METHOD OF PREPARATION OF THE REDUCED ADSTRINGENCE OF VEGETABLE PROTEIN PRODUCTS, VEGETABLE PROTEIN P

专利摘要:
abstract “production of legume protein products with reduced astringency” pulse proteins of reduced astringency are obtained by fractionating legume protein products that are completely soluble and heat stable in aqueous media with an acidic pH value of less than approx. of 4.4 in lower molecular weight, less astringent proteins and higher molecular weight, more astringent proteins.
公开号:BR112015029903A2
申请号:R112015029903
申请日:2014-05-30
公开日:2020-04-28
发明作者:I Segall Kevin；Schweizer Martin；Medina Sarah
申请人:Burcon Nutrascience Mb Corp；
IPC主号:

专利说明:

METHOD OF PREPARATION OF THE REDUCED ADSTRINGENCE OF VEGETABLE PROTEIN PRODUCTS, VEGETABLE PROTEIN PRODUCTS AND AQUEOUS SOLUTION
Related Order Reference
[0001] This application claims priority under 35 USC 119 (e) of US Provisional Patent Applications Nos. 61 / 828,735 deposited on May 30, 2013 and 61 / 927,182 deposited on January 14, 2014.
Field of the Invention
[0002] The present invention relates to the production of legume protein products, preferably isolated from legume protein.
Background of the Invention
[0003] In North American Patent Applications No. 13 / 103,528 filed on May 9, 2011 (North American Patent Publication No. 2011-027497 published on November 10, 2011), 13 / 289,264 filed on 4 November 2011 (United States Patent Publication No. 20120135117 published on May 31, 2012), 13 / 556,357 filed on July 24, 2012 (United States Patent Publication No. 2013-0189408 published on July 25, 2012 2013) and 13 / 642,003 filed on January 7, 2013 (United States Patent Publication No. 2013-0129901 published on May 23, 2013), attributed to this assignee and the disclosures of which are incorporated into this document by reference, the provision of a new legume protein product is described which has a protein content of at least about 60% by weight (N x 6.25) on a dry weight basis, preferably a legume protein isolate which has a protein content of hair
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2/72 minus about 90% by weight (N x 6.25) d.b .. The legume protein product has a unique combination of properties, namely:
- completely soluble in aqueous media at acid pH values of less than about 4.4;
- heat-stable in aqueous media at acid pH values of less than about 4.4;
- does not require stabilizers or other additives to keep the protein product in solution;
- is low in phytic acid;
- does not require enzymes in its production.
[0004] This new protein product from legumes is prepared by a method comprising:
(a) extracting a source of legume protein with an aqueous solution of calcium salt, preferably an aqueous solution of calcium chloride, to cause the legume protein to solubilize from the protein source and to form an aqueous solution of legume protein , (b) separating the aqueous vegetable protein solution from the residual vegetable protein source, (c) optionally diluting the aqueous vegetable protein solution, (d) adjusting the pH of the aqueous vegetable protein solution to a pH of about 1.5 to about 4.4, preferably about 2 to about 4, to produce an acidified solution of legumes protein, (e) optionally clarifying the acidified solution of legumes protein if it is not yet clear, ( f) alternatively from steps (b) to (e),
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3/72 optionally dilute and then adjust the pH of the combined aqueous legume protein solution and residual legume protein source to a pH of about 1.5 to about 4.4, preferably about 2 to about 4 , then separate the acidified legume protein solution, preferably clear, from the residual legume protein source, (g) optionally concentrate the aqueous legume protein solution while maintaining ionic resistance substantially constant by a selective membrane technique, (h ) optionally diafiltrate the protein solution of
legumes optionally concentrated, and (i) optionally dry the solution in protein from legumes optionally concentrated and optionally diafiltered. [0005] 0 protein product in legumes
preferably it is an isolate that has a protein content of at least about 90% by weight, preferably at least about 100% by weight (N x 6.25) d.b ..
[0006] In certain acidic drinks, particularly those that have a pH at the low end of the acceptable pH range for acidic drinks, the new protein product from legumes tends to induce an unpleasant astringent feeling in the mouth.
Summary of the Invention
[0007] Now, it has been discovered that this undesirable astringency can be reduced or eliminated by modifying the procedure used to manufacture the new protein product from legumes.
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[0008] In accordance with the present invention, a method of preparing the legume protein product with reduced astringency is provided, comprising:
(a) extracting a protein source from legumes with an aqueous solution of calcium salt to cause solubilization of the protein from the protein source and to form an aqueous protein solution from legumes, (b) separating the aqueous protein solution from Legumes from the residual legume protein source, (c) Optionally dilute the aqueous legume protein solution, (d) Adjust the pH of the aqueous legume protein solution to a pH of about 1.5 to about 4.4 to produce an acidified legume protein solution, (e) optionally clarify the acidified legume protein solution, if not already clear, (f) alternatively from steps (b) to (e), optionally dilute and then adjusting the pH of the combined aqueous protein protein solution and the residual protein protein source to a pH of about 1.5 to about 4.4 and then separating the acidified protein solution, preferably clear, from the source of residual legume protein, and (g) fractionating the proteins in the acidified legume protein solution to separate the lower molecular weight, less astringent proteins of higher molecular weight, more astringent proteins.
[0009] In accordance with one aspect of the present invention, the
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The process is modified to remove proteins that precipitate at a pH of about 5 to about 6.5 and that can interact with salivary proteins, thereby producing a less astringent product. In order to precipitate the protein fraction, the pH of the acidified protein protein solution, preferably after concentration and partial diafiltration, is adjusted to about 5 to about 6.5, preferably about 5.5 to about 6, 0. The precipitated protein is removed and the protein remaining in the solution is then reacidified to about pH 3 and another membrane processed to form one of the products of the invention. The collected material that precipitates by adjusting the pH can also be a process for providing another product of the invention. The precipitated material can be processed as follows:
1. Optionally washed with water and spray dried at about pH 5.5; or
2. Optionally washed with water, adjusted to a pH of about 6 to 8, then spray dried; or
3. Adjusted to about pH 3, processed membrane then spray dried; or
4. Adjusted to about pH 3, processed membrane, adjusted to a pH of about 6 to 8 then spray-dried.
[0010] This product is typically intended for use in neutral applications.
[0011] The less astringent proteins that remain in solution when the precipitation method mentioned above is applied appear to be of lower molecular weight than the more astringent species. In another aspect of this
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6/72 invention, the more astringent protein component can be separated from the less astringent protein component by membrane processing. The optional concentration and diafiltration of a protein solution containing a mixture of the most and least astringent proteins using a membrane with an appropriate pore size allows smaller, less astringent proteins to pass through the permeate, while retaining the most astringent species in the solution concentrated protein. The less astringent proteins can be separated from the contaminants by a subsequent ultrafiltration and / or diafiltration step using a membrane that has a smaller pore size than that employed in the fractionation step. The purified least astringent protein fraction is a product of the invention. The solution of the broadest, most astringent protein species can also be further processed and optionally neutralized to form another product of the invention, which is typically intended for use in neutral applications.
[0012] According to a further aspect of the present invention, a protein product of legumes is provided which has a protein content of at least about 60% by weight (N x 6.25) d.b. is that:
- it is completely soluble in aqueous media at acid pH values of less than 4.4;
- it is heat-stable in aqueous media at acid values of less than about 4.4;
- does not require stabilizers or other additives to keep the protein product in solution or suspension;
- is low in phytic acid;
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- does not require enzymes in its production;
- is low in astringency when tested in an aqueous solution at a pH below about 5.
[0013] The protein product of legumes preferably has a protein content of at least about 90% by weight, more preferably of 100% by weight, (N x 6.25) db. The protein product of legumes preferably does not it is hydrolyzed and preferably has a phytic acid content of less than about 1.5% by weight, preferably less than about 0.5% by weight.
[0014] According to a further aspect of the present invention, a protein product of legumes is provided which has a protein content of at least about 60% by weight (N x 6.25) d.b. and which has low astringency when tested in an aqueous solution at a pH below about 5 which is substantially completely soluble in an aqueous medium at a pH of less than about 4.4.
[0015] The legume protein product can be mixed with water-soluble powdered materials to produce aqueous solutions of the mixture, preferably a powdered drink. The legume protein product can be formed with an aqueous solution, such as a drink, which is heat-stable at a temperature of less than about 4.4. The drink can be a clear drink in which the dissolved legume protein product is completely soluble and transparent, or it can be a non-transparent drink in which the dissolved legume protein increases or fails the level of fog or cloud.
[0016] In accordance with a further aspect of the present invention, a protein product of legumes is provided
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8/72 that has a molecular weight profile as determined
by methods described at the Example 25, which is: fence in 10 The fence in 75% greater than about in 100,000 Gives;fence in 10 The fence in 45% of about 15. 000 about
of 100,000 Da;
fence in 8 the fence 55% of about 5,000 The fence in 15 .000 Da;fence in 2 the fence 12% of about 1,000 The fence in 5. 000 Da.[0017] 0 profile of molecular weight can to be: fence in 15 to about 40 % bigger then fence in 100. 000
Da about 25 to about 40% of about 15,000 to about 100,000 Da;
about 15 to about 50% of about 5,000 to about 15,000 Da;
about 3 to about 10% of about 1,000 to about 5,000 Da.
[0018] In accordance with another aspect of the present invention, a protein product of legumes is provided which has a molecular weight profile, as determined by the methods described in Example 25, which is:
about 10 to about 85% greater than about 100,000 Da;
about 10 to about 45% of about 15,000 to about 100,000 Da;
about 0 to about 40% of about 5,000 to about 15,000 Da;
about 1 to about 34% of about 1,000 to about
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5,000 Da.
[0019] The molecular weight profile can be:
about 18 to about 78% greater than about 100,000 Da;
about 15 to about 38% of about 15,000 to about 100,000;
Da about 2 to about 35% of about 5,000 to about 15,000 Da;
about 3 to about 25% of about 1,000 to about 5,000 Da.
[0020] Accordingly, with a further aspect of the present invention, a protein product of legumes is provided which has a protein content of at least about 60% by weight (N x 6.25) d.b. which has a solubility of 1% w / v protein in water at a pH of about 2 to about 7 of more than about 50%, as determined by the methods described in Example 5. The protein product of legumes is preferably it has a protein content of at least about 90% by weight, more preferably at least about 100% by weight (N x 6.25) db.
[0021] The less astringent legume protein products of the invention, produced according to the processes in this document, are suitable, not only for protein fortification of acidic media, but can be used in a wide variety of conventional applications of the products protein, including, but not limited to, protein fortification of processed foods and beverages, oil emulsification and as a foaming agent in products that retain gases. Legume protein products can also be used in
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10/72 nutritional supplements. Legume protein products can also be used in dairy and alternative dairy products that are mixtures of dairy / vegetable ingredients. Other uses of legume protein products are in animal feed, animal feed and in industrial and cosmetic applications and in personal care products.
General Description of the Invention
[0022] The initial step in the process of supplying legume protein products involves solubilizing legume protein from a legume protein source. The pulses to which the invention can be applied include, but are not limited to, lentils, chickpeas, dried peas and dried beans. The protein source of pulses can be pulses or any pulse product or by-product derived from pulse processing. For example, the protein source of legumes can be a flour prepared by grinding an optionally peeled pulse. As another example, the source of legume protein may be a protein-rich pulse fraction formed by peeling and shredding a pulse and then air classification of the peeled and crushed material into starchy and protein-rich fractions. The legume protein product recovered from the legume protein source may be the naturally occurring protein in pulses, or the protein material may be a protein modified by genetic manipulation, but which has hydrophobic characteristics and polar properties of the natural protein.
[0023] Protein solubilization from protein source
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11/72 of legume material is most conveniently made using calcium chloride solution, although solutions of other calcium salts can be used. In addition, other alkaline earth metal compounds can be used, such as magnesium salts. Also, the extraction of legume protein from the legume protein source can be carried out using calcium salt solution in combination with another salt solution, such as sodium chloride. Additionally, the extraction of the legume protein from the legume protein source can be carried out using water or another salt solution, such as sodium chloride, with calcium salt being subsequently added to the aqueous legume protein solution produced in the extraction step. . The precipitate formed by adding the calcium salt is removed before further processing.
[0024] As the concentration of the calcium salt solution increases, the degree of solubilization of the protein from the legume protein source initially increases until it reaches a maximum value. Any subsequent increase in salt concentration does not increase the total solubilized protein. The concentration of the calcium salt solution that causes maximum protein solubilization varies depending on the salt in question. It is generally preferred to use a concentration value of less than about 1.0 M, and more preferably a value of about 0.10 to about 0.15 M.
[0025] In a batch process, the salt solubilization of the protein is carried out at a temperature of about 1 ° C to about 100 ° C, preferably about 15 ° C to about
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65 ° C, more preferably about 20 ° C to about 35 ° C, preferably accompanied by stirring to decrease the solubilization time, which is generally about 1 to about 60 minutes. It is preferred to perform solubilization to extract substantially as much protein from the legume protein source as is practicable, in order to provide a high total product yield.
[0026] In a continuous process, the extraction of protein from the protein source of legumes is carried out in any way consistent with the effectiveness of a continuous extraction of protein from the protein source of legumes. In one embodiment, the protein source of legumes is continuously mixed with the calcium salt solution and the mixture is transported through a tube or conduit that has a length and flow rate for a sufficient residence time to effect the desired extraction according to the parameters described in this document. In such a continuous procedure, the salt solubilization step is carried out in a time of about 1 minute to about 60 minutes, preferably to effect the solubilization to extract substantially as much protein from the legume protein source as is practicable. Solubilization in the continuous procedure is carried out at temperatures between about 1 ° C and about 100 ° C, preferably between about 15 ° C and about 65 ° C, more preferably between about 20 ° and about 35 ° C.
[0027] Extraction is generally carried out at a pH of about 4.5 to about 11, preferably about 5 to about 7. The pH of the extraction system (source of legume protein and calcium salt solution) Can be
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13/72 adjusted to any desired value within the range of about 4.5 to about 11 for use in the extraction step by using any convenient food grade acid, usually hydrochloric acid or phosphoric acid, or food grade alkali, usually sodium hydroxide as required.
[0028] The concentration of the protein source of legumes in the calcium salt solution during the solubilization step can vary widely. Typical concentration values are about 5 to about 15% w / v.
[0029] The protein extraction step with the aqueous salt solution has the additional effect of solubilizing fats that may be present in the legume protein source, which then results in the fats that are present in the aqueous phase.
The protein solution that results from the extraction step generally has a protein concentration of about 5 to about 50 g / L, preferably about 10 to about 50 g / L.
[0031] The aqueous solution of calcium salt may contain an antioxidant. The antioxidant can be any convenient antioxidant, such as sodium sulfite or ascorbic acid. The amount of antioxidant employed can vary from about 0.01 to about 1% by weight of the solution, preferably about 0.05% by weight. The antioxidant serves to inhibit the oxidation of any phenolics in the protein solution.
[0032] The aqueous phase resulting from the extraction step can then be separated from the residual legume protein source, in any convenient way, such as using a decanter centrifuge, followed by
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14/72 disk centrifugation and / or filtration, to remove material from the residual legume protein source. The separation step can be conducted at any temperature within the range of about 1 ° C to about 100 ° C, preferably about 15 ° to about 65 ° C, more preferably about 20 ° C to about 35 ° Ç. Alternatively, the optional dilution steps and acidification steps described below can be applied to the mixture of the aqueous legume protein solution and residual legume protein source, with subsequent removal of material from the residual legume protein source by the separation step described above. The separate residual legume protein source can be dried for disposal or further processed, such as recovering residual amide and / or protein. The residual protein can be recovered by reextracting the separated residual legume protein source with fresh calcium salt solution and the protein solution produced by clarification combined with the initial protein solution for further processing as described below. Alternatively, the separate residual legume protein source can be processed by a conventional precipitation process or any convenient procedure to recover the residual protein.
[0033] The aqueous protein solution of legumes can be treated with an antifoam, such as any non-silicone based antifoam, of adequate food quality, to reduce the volume of the foam formed by further processing. The amount of defoamer used is generally greater than about
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0.0003% w / v. Alternatively, the defoamer in the
amount described can be added in the steps in extraction.[0034] The solution watery protein of legumes separated Can be submitted to a operation in degreasing, if required, according described in Patents North American No. 5,844,086 and 6,005. 076, assigned to the present assignment io and the disclosures of
which are incorporated into this document by reference. Alternatively, degreasing the separated aqueous protein protein from legumes can be achieved by any other convenient procedure.
[0035] The aqueous protein solution of legumes can be treated with an absorbent, such as powdered activated carbon or granulated activated carbon, to remove color and / or odor compounds. Such an absorbent treatment can be carried out under any convenient conditions, generally at room temperature of the separated aqueous protein solution. For powdered activated carbon, an amount of about 0.025% to about 5% w / v, preferably about 0.05% to about 2% w / v, is employed. The absorption agent can be removed from the protein solution of legumes by any convenient means, such as by filtration.
[0036] The resulting aqueous protein solution of legumes can be diluted generally with about 0.1 to about 10 volumes, preferably about 0.5 to about 2 volumes of aqueous diluent, in order to decrease the conductivity of the solution aqueous protein from legumes to a value of generally below about 105 mS,
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16/72 preferably about 4 to about 21 mS. Such dilution is generally carried out using water, although the diluted salt solution, such as sodium chloride or calcium chloride, which has a conductivity up to about 3 mS, can be used.
[0037] The diluent, with which the legume protein solution is mixed, generally has the same temperature as the legume protein solution, but the diluent can have a temperature of about 1 ° C to about 100 ° C , preferably about 15 ° C to about 65 ° C, more preferably about 20 ° C to about 35 ° C.
[0038] The optionally diluted legume protein solution is then adjusted in pH to a value of about 1.5 to about 4.4, preferably about 2 to about 4, by the addition of any suitable food-grade acid , such as hydrochloric acid or phosphoric acid, to result in an acidified aqueous protein protein solution, preferably a clear acidified aqueous protein protein solution. The aqueous acidified legume protein solution has a conductivity generally below about 110 mS for a diluted legume protein solution, or generally below about 115 mS for an undiluted legume protein solution, in both cases preferably about 4 to about 26 mS.
[0039] As mentioned above, as an alternative to the previous separation of the residual legume protein source, the aqueous legume protein solution and the material of the residual legume protein source can be optionally diluted and acidified together and then
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17/72 aqueous acidified legume protein solution is clarified and separated from the residual legume protein source material by any technique as discussed above. The aqueous solution of acidified legume protein can optionally be defatted, optionally treated with an absorbent and optionally treated with a defoamer, as described above.
[0040] The aqueous solution of acidified legume protein can be subjected to a heat treatment to inactivate the thermal unstable antinutritional factors, such as trypsin inhibitors, present in such a solution as a result of the extraction of material from the legume protein source during the extraction step. Such a heating step also provides the benefit of reducing the microbial load. Generally, the protein solution is heated to a temperature of about 70 ° C to about 160 ° C, preferably about 80 ° C to about 120 ° C, more preferably about 85 ° to about 95 ° C , for about 10 seconds to about 60 minutes, preferably about 10 seconds to about 5 minutes, more preferably about 30 seconds to about 5 minutes. The heat treated acidified legume protein solution can be cooled for further processing, as described below, to a temperature of about 2 ° C to about 65 ° C, preferably about 50 ° C to about 60 ° C.
[0041] If the optionally diluted, acidified and optionally heat treated legume protein solution is not transparent, it can be clarified by any convenient procedure such as filtration or
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18/72 centrifugation.
[0042] According to one aspect of the present invention, the acidified aqueous protein protein solution, preferably after the concentration and diafiltration steps described below, more preferably after performing the partial concentration and diafiltration steps described below, is adjusted to pH for the range of about 5 to about 6.5, preferably about 5.5 to about 6.0 to effect protein precipitation and fractionation. Such pH adjustment can be carried out using any suitable food grade alkali, such as aqueous sodium hydroxide solution. The protein that precipitates at that pH is collected by any convenient means such as centrifugation and the resulting solution is reacidified to a pH of about 1.5 to about 4.4, preferably about 2 to about 4, by adding any suitable food grade acid, such as hydrochloric acid or phosphoric acid, to result in a reacidified aqueous solution of legume protein, preferably a clear reacidified aqueous solution of legume protein. This aqueous solution of reacidified legume protein contains the least astringent protein species. The aqueous solution of reacidified legume protein is then processed according to the steps described below.
[0043] The protein precipitated from about pH 5 to about 6.5 and separated from the resulting solution can be further processed. The precipitate, which is the most astringent protein fraction, can be washed with water and dried by any convenient procedure such as
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19/72 drying for spraying or freeze drying. Alternatively, the precipitate can be washed with water, adjusted to a pH of about 6 to 8 and then dried. The precipitate can be adjusted to a pH of about 1.5 to about 4.4, preferably about 2 to about 4, then the membrane is processed as described below and dried. The precipitate can be adjusted to a pH of about 1.5 to about 4.4, preferably about 2 to about 4, the membrane processed as described below, adjusted to a pH of about 6 to about 8, and then dried.
[0044] The aqueous solution of acidified legume protein can be concentrated before fractionation by adjusting the pH as described above. Such a concentration step increases the protein concentration of the solution while maintaining its ionic resistance substantially constant. Such a concentration step is generally carried out to provide a concentrated protein solution of legumes that has a protein concentration of about 50 to about 300 g / L, preferably about 100 to about 200 g / L. When the acidified aqueous protein solution is partially concentrated before precipitation and removal of the most astringent protein at a pH of about 5 to about 6.5, the concentration step is preferably carried out at a protein concentration below about 50 g / L. The concentrated or partially concentrated acidified aqueous solution can be diluted with water before the pH adjustment step in order to reduce the sample viscosity and facilitate the recovery of the precipitated protein by adjusting the pH.
[0045] The aqueous protein solution of legumes
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Reacidified 20/72 can also be concentrated to increase its protein concentration while keeping its ionic resistance substantially constant. Such a concentration step is generally carried out to provide a concentrated reacidified legume protein solution that has a protein concentration of about 10 to about 300 g / L, preferably about 100 to about 200 g / L. When the reacidized aqueous protein solution is partially concentrated, the concentration step is preferably carried out at a protein concentration of less than about 10 g / L.
[0046] Such concentration steps may be carried out in any convenient manner consistent with batch or continuous operation, such as employing any convenient selective membrane technique, such as ultrafiltration or diafiltration, using membranes such as hollow fiber membranes or spiral wound, with a suitable molecular weight cut, such as about 1,000 to about 1,000,000 daltons, preferably about 1,000 to about 100,000 daltons, more preferably about 1,000 to about 10,000 daltons having regard to the different membrane materials and configurations, and, for continuous operation, sized to allow the desired degree of concentration as the aqueous protein solution passes through the membranes.
[0047] As is well known, ultrafiltration and similar selective membrane techniques allow low molecular weight species to bypass them while preventing higher molecular weight species from doing so. Low molecular weight species include
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21/72 only the ionic species of the salt, but also low molecular weight materials extracted from the source material, such as carbohydrates, pigments and low molecular weight proteins including the less astringent proteins (discussed below) and the anti-nutritional trypsin inhibitors. The molecular weight cut of the membrane is normally chosen to ensure the retention of a significant proportion of the protein in the solution, while allowing contaminants to pass through, in relation to different membrane materials and configurations.
[0048] The protein solution of concentrated acidified or reacidified concentrated legumes can be subjected to a diafiltration step using water or a diluted saline solution. The diafiltration solution can be at its natural pH or at a pH equal to that of the protein solution being diafiltered or at any intermediate pH value. Such diafiltration can be carried out using about 1 to about 40 volumes of diafiltration solution, preferably about 2 to about 25 volumes of diafiltration solution. In the diafiltration operation, additional amounts of contaminants are removed from the aqueous protein solution of legumes by passing through the membrane with the permeate. This purifies the aqueous protein solution and can also reduce viscosity. The diafiltration operation can be carried out until no significant additional amounts of contaminants or visible color are present in the permeate or, in the case of the reacidified protein solution, until the retentate has been sufficiently purified so that, when dried, it provides an isolate of legume protein
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with a content of protein of hair any less fence 90% by weight (N x 6.25) d.b .. Such diafiltration can to be effected using the same membrane like for The step of concentration However, if wanted, The step of diafiltration Can be effected using an membrane
separated with a different molecular weight cut, such as a membrane that has a molecular weight cut in the range of about 1,000 to about 1,000,000 daltons, preferably about 1,000 to about 100,000 daltons, more preferably about 1,000 to about 10,000 daltons with respect to different membrane materials and configuration.
[0049] Alternatively, the diafiltration step can be applied to the acidified or reacidified aqueous protein solution prior to concentration or to the acidified partially concentrated or partially concentrated acidified aqueous protein solution. Diafiltration can also be applied at multiple points during the concentration process. When diafiltration is applied before concentration or to the partially concentrated solution, the resulting diafiltered solution can then be fully concentrated. The reduction in viscosity achieved by multi-time diafiltration, as the protein solution is concentrated, can allow for a higher final protein concentration, fully concentrated to be achieved. In the case of the reacidified protein solution, this reduces the volume of the material to be dried.
[0050] An antioxidant may be present in the diafiltration medium during at least part of the
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23/72 diafiltration. The antioxidant can be any convenient antioxidant, such as sodium sulfite or ascorbic acid. The amount of antioxidant used in the diafiltration medium depends on the materials used and can vary from about 0.01 to about 1% by weight, preferably about 0.05% by weight. The antioxidant serves to inhibit the oxidation of any phenolics present in the concentrated legume protein solution.
[0051] Concentration steps and optional diafiltration steps can be performed at any convenient temperature, usually about 2 ° C to about 65 ° C, preferably about 50 ° C to about 60 ° C, and for a period of time to effect the desired degree of concentration. The temperature and other conditions used to some degree depend on the membrane equipment used to perform the membrane processing, the desired protein concentration in the solution and the effectiveness of removing contaminants into the permeate.
[0052] According to another aspect of this invention, the concentration and optional diafiltration steps are operated in the aqueous solution of leguminous protein acidified in such a way as to separate the lower molecular weight proteins, less astringent from the higher molecular weight proteins , more astringent. When this process is employed, the molecular weight cut of the concentration and diafiltration membranes is chosen to allow smaller, less astringent proteins to pass through the permeate with the contaminants. Such concentration and diafiltration steps can be carried out in any convenient manner consistent with batch operation or
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24/72 continues, as employing any convenient selective membrane technique, such as microfiltration or ultrafiltration, using membranes, such as hollow fiber membranes or spiral wound membranes, with an appropriate molecular weight cut, such as about 0 , 05 to about 0.1 pm, preferably about 0.08 to about 0.1 pm for microfiltration and about 10,000 to about 1,000,000 daltons, preferably about 100,000 to about 1,000,000 daltons for ultrafiltration, having different materials and membrane configurations, and, for continuous operation, dimensioned to allow the desired degree of concentration as the aqueous protein solution passes through the membranes. In the concentration step, the acidified protein solution is concentrated to a protein concentration of about 50 to about 300 g L, preferably about 100 to about 200 g / L. The concentrated protein solution can then be diafiltered with water or diluted saline. The diafiltration solution can be at its natural pH or at a pH equal to that of the protein solution that is diafiltered or any intermediate pH value. Such diafiltration can be carried out using about 1 to about 40 volumes of diafiltration solution, preferably about 2 to about 25 volumes of diafiltration solution. Optional concentration and diafiltration steps can be carried out at any convenient temperature, generally from about 2 ° C to about 65 ° C, preferably about 50 ° to about 60 ° C. Smaller, less astringent proteins are captured in the permeate of membrane processes alongside other small molecule contaminants.
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[0053] The less astringent proteins are then separated from the contaminants by subsequent concentration of the protein solution (permeate from step 1) by processing the membrane such as ultrafiltration to a
concentration of protein of fence in 10 to about 300 g / L, preferably about of 100 The about 200 g / L e diafiltration optional. When The solution of protein (permeated by step 1) is partially and concentrated, the stage
concentration is preferably carried out at a protein concentration of less than about 10 g / L. The concentration and diafiltration steps are performed using a membrane that has a lower molecular weight cut such as about 1,000 to about 100,000 daltons, preferably 1,000 to about 10,000 daltons operated as described above.
[0054] Additional products can be obtained from the retentation of the membrane fractionation process, which contains the most astringent proteins. This protein solution can be dried by any means
convenient, with or without adjustment of pH of the solution in protein from about 6 to about 8 using alkali in food quality. [0055] The concentration and the phases optional in
diafiltration used in the purification of aqueous solutions of less astringent proteins derived either from precipitation or from the membrane fractionation procedure can be carried out in this document in such a way that the less astringent legume protein product recovered contains less than about 90% by weight protein (N x 6.25) db, such as at least about
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26/72 of 60% by weight of protein (N x 6.25) d.b .. By partially concentrating and / or partially diafiltering the aqueous protein solution from legumes, it is only possible to partially remove contaminants. That protein solution can then be dried to provide a protein product from legumes with lower levels of purity. The legume protein product is highly soluble and capable of producing less astringent protein solutions, preferably clear, less astringent protein solutions, under acidic conditions.
[0056] As mentioned earlier, the pulses contain antinutritional trypsin inhibitors. The level of trypsin inhibitor activity in the final legume protein product can be controlled by manipulating several process variables.
[0057] As noted above, the heat treatment of the aqueous acidified protein protein solution can be used to inactivate heat-unstable trypsin inhibitors. The partially concentrated or fully concentrated acidified legume protein solution can also be heat treated to inactivate heat unstable trypsin inhibitors. Such heat treatment can also be applied to the reacidified legume protein solution that arises from the precipitation fractionation method or the less astringent protein solution, of lower molecular weight that arise from the membrane separation method, before or after partial concentration or complete. When the heat treatment is applied to a solution that is no longer fully concentrated, the resulting heat treated solution can then be
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27/72 additionally concentrated.
[0058] Acidifying and processing the legume protein solution membrane at a lower pH, such as 1.5 to 3, can reduce the activity of the trypsin inhibitor compared to processing the solution at a higher pH, such as 3 to 4 , 4. When the protein solution is concentrated and diafiltered at the low end of the pH range, it may be desired to raise the pH of the retentate before drying. The pH of the concentrated and diafiltered protein solution can be raised to the desired value, for example, pH 3, by adding any suitable food grade alkali, such as sodium hydroxide.
[0059] In addition, a reduction in trypsin inhibitor activity can be achieved by exposing the pulse materials to reduce the agents that affect or reorganize the disulfide bonds of the inhibitors. Suitable reducing agents include sodium sulfite, cysteine and Nacetyl cysteine.
[0060] The addition of such reducing agents can be carried out at various stages of the total process. The reducing agent can be added with the legume protein source material in the extraction step, can be added to the clarified aqueous protein protein solution after removal of the residual legume protein source material, can be added to the retentate optionally diafiltered before drying or can be mixed dry with dry legume protein product. The addition of the reducing agent can be combined with the heat treatment step and membrane processing steps, as described above.
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[0061] If it is desired to retain the active trypsin inhibitors in the products, this can be achieved by eliminating or reducing the intensity of the heat treatment step, not using reducing agents, operating the concentration and diafiltration steps at the upper end of the pH, such as 3 to 4.4.
[0062] Any of the concentrated and optionally diafiltered protein solutions described above can be subjected to an additional degreasing operation, if required, as described in US Patent Nos. 5,844,086 and 6,005,076. Alternatively, degreasing of concentrated and optionally diafiltered protein solutions can be achieved by any other convenient procedure.
[0063] Any of the concentrated and optionally diafiltered aqueous protein solutions described above can be treated with an absorbent, such as powdered activated carbon or granulated activated carbon, to remove color and / or odor compounds. Such an absorbent treatment can be carried out under any convenient conditions, generally at room temperature of the concentrated protein solution. For powdered activated carbon, an amount of about 0.025% to about 5% w / v, preferably about 0.05% to about 2% w / v, is employed. The absorbent can be removed from the protein solution of legumes by any convenient means, such as by filtration.
[0064] The concentrated and optionally diafiltered aqueous protein solutions of collected legume protein described above can be dried by any convenient technique, such as
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29/72 spray drying or lyophilization. A pasteurization step can be carried out on legume protein solutions or on resuspended legume protein precipitates before drying. Such pasteurization can be carried out under any desired pasteurization conditions. Generally, the concentrated and optionally diafiltered legume protein solution or resuspended legume protein precipitate is heated to a temperature of about 55 ° C to about 70 ° C, preferably about 60 ° C to about 65 ° C, for about 30 seconds to about 60 minutes, preferably about 10 minutes to about 15 minutes. The pasteurized concentrated legume protein solution or resuspended legume protein precipitate can then be cooled by drying, preferably at a temperature of about 25 ° C to about 40 ° C.
[0065] Each of the dried legume protein products obtained by the procedures described above has a protein content greater than about 60% by weight. Preferably, the dried legume protein products are isolated with a protein content in excess of about 90% by weight of protein, preferably at least about 100% by weight, (N x 6.25) d.b ..
[0066] The less astringent legume protein products produced in this document are soluble in an acidic aqueous environment, making the products ideal for incorporation into drinks, both carbonated and non-carbonated, to provide protein fortification to them. Such drinks have a wide range of acidic pH values, ranging from about 2.5 to about 5. Products
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30/72 of legume protein provided in that document can be added to such drinks in any amount convenient to provide protein fortification for such drinks, for example, at least about 5 g of legume protein per serving. The legume protein product added dissolves in the drink and the haze or cloud level is not increased by thermal processing. The protein product of legumes can be mixed with dry drink before reconstituting the drink by dissolving it in water. In some cases, modification to the normal beverage formulation to tolerate the composition of the invention may be necessary where the components present in the beverage may adversely affect the ability of the composition of the invention to remain dissolved in the beverage.
Examples
Example 1:
[0067] This Example illustrates the production of the legume protein product with reduced astringency of the invention using methods where the acidified legume protein solution is partially concentrated or concentrated and diafiltered before the precipitation of the most astringent protein by adjusting the pH.
[0068] 'a' kg of 'b' was combined with 'c' L of purified water by reverse osmosis (RO) and the mixture stirred for 'd' minutes at room temperature. The insoluble material was removed and the sample partially clarified by centrifugation, yielding a protein solution having a protein concentration of 'e'% by weight. To this protein solution was added 'f kg of chloride stock solution
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31/72 of calcium, prepared by dissolving 1 kg of calcium chloride pellets (95.5%) by 9 L of water 'g'. The insoluble material was removed and the sample clarified by centrifugation, yielding 'h' L of protein extract solution having a wt% protein concentration.
'j' L of protein extract solution was combined with 'k' L of water by RO and the pH of the sample decreased to T with HCl solution (concentrated HCl diluted with an equal volume of water), 'm' L of acidified protein solution was clarified by performing it in a microfiltration system equipped with a Membralox ceramic membrane with a pore size of 0.8 pm operated at 'n' ° C to 'o' L permeate (protein solution acidified
clarified) be collected, ' P' L for 'q', by having a content in protein from 'r'% in Weight was 's' concentrated up to 't' L using a membrane in ultrafiltration PES by having one
pore size of 1,000 daltons operated at a temperature of around 'u' ° C. 'v' L of 'w' concentrated protein solution was then diafiltered with 'x' L of water by RO at about 'y' ° C to provide 'z' of 'aa' diafiltered concentrated protein solution having a content % ab protein by weight. The 'ac' solution of concentrated diafiltered protein was diluted with 'ad' L water by RO and the pH adjusted to 'ae' with NaOH solution, which caused the formation of a precipitate, 'af' kg of wet precipitate was removed by centrifugation to provide 'ag' L of protein solution with a protein content of 'ah'% by weight. The pH of the protein solution was reduced to 'ai' and then 'aj' L of the reacidified protein solution was polished by running the solution through a membrane of
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32/72 Membralox ceramic microfiltration having a pore size of 0.80 pm and operated at 'ak' ° C until 'al' L of permeate is collected, 'am' L of 'an' was then reduced in volume to 'to' L by concentration on a PES ultrafiltration membrane having a pore size of 1,000 daltons operated at a temperature of about 'ap' ° C. The resulting 'aq' concentrated protein solution, having a 'air' wt% protein content was then diafiltered with 'as' L of water by RO at about 'at' ° C 'au' to provide 'av' 'kg of concentrated diafiltered protein solution having a protein content of' aw '% by weight. This represented a yield of 'ax'% of the protein in the protein extract solution resulting from the clarification step after the addition of calcium chloride, 'ay' kg of concentrated diafiltered protein solution was spray dried to yield a protein product , having a protein content of 'az'% (N x 6.25) db, called 'ba' 'bb'.
[0069] The 'af kg of wet precipitate collected, having a protein content of' be ', represented a yield of' bd '% of the protein in the protein extract solution resulting from the clarification step after the addition of calcium chloride , 'be' kg of this precipitate was diluted with 'bf kg of water following the pH adjusted to' bg 'and the mixture pasteurized in about' bh 'for' bi 'minutes. The 'bj' sample was then spray dried to provide a dry protein product having a protein content of 'bk'% (N x 6.25) d.b. which was called 'ba' 'bl'.
[0070] The parameters 'a' to 'bl' are determined in the following table 1.
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Table 1 - Parameters for the production of protein products by the precipitation and fractionation method
ba YP20-D23-13A YP20-D24-13A YP20-E02-13A LE03-D02-14A The 30 30 60 36 B Yellow pea protein concentrate Yellow pea protein concentrate Yellow pea protein concentrate Lentil flourwhole green ç 500 500 1000 600 d 30 30 30 10 and 2.69 2.68 2.67 1.27 f 63.14 65 137.34 80 g And the stirred mixture15 minutes And the stirred mixture15 minutes And the mixture stirred 15 minutes AT H 459 484 978 586 i 1.60 1.41 1.55 0.68 j 459 484 978 586 k 371 317 640 368 1 2.91 3.12 3.00 3.02 m 830 790 AT AT n 59 59 AT AT 0 NR NR AT AT P 780 700 1585 975 q Clarified acidified protein solution Clarified acidified protein solution Clarified acidified protein solution Clarified acidified protein solution r 0.81 0.74 0.81 0.40 s Partially AT AT Partially t 120 72 215 50 u 57 57 58 58 V 120 72 215 50
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w Partially AT AT Partially X 240 144 430 100 y 60 61 59 60 z 120 L 72 L 220 L 48.56 kg aa Partially AT AT AT ab 4.04 5.57 5.62 5.13 B.C Partially AT AT AT ad 120 78 344 NR ae 5.63 5.73 About 5.5 6.10 af 33.50 31.12 105.36 16.14 ag 230.1 128.5 444 80 ah 0.40 0.51 0.36 0.65 there 3.08 2.79 3.11 2.99 aj AT AT AT 80 ak AT AT AT 46 al AT AT AT 64 am 230 150 444 64 an Reacidified protein solution Reacidified protein solution Reacidified protein solution Reacidified protein solution to 78 25 32.5 22 ap 58 52 54 58 aq AT AT AT Partially air 1.16 1.91 4.62 0.62 at 78 25 32.5 22 at 60 59 60 59 au And then stillfocused AT And then still concentrated AT av 34.56 29.14 24.86 21.00 a W 2.87 2.38 6.25 1.51
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ax 13.5 10.1 10.2 8.0 ay 35.54 29.14 24.86 21.00 az 100.17 99.36 101.84 92.26 bb YP705 YP705 YP705 LE705 bc 12.33 11.65 10.38 11.18 bd 56.3 53.2 72.2 45.2 be 8.5 8.94 24 16.14 bf 8.5 8.94 0 8.00 bg 7.07 6.82 AT AT bh AT AT AT 66 bi AT AT AT 15 bj AT AT AT Pasteurized bk 102.58 102.49 101.44 102.08 bl YP705P YP705P YP705P LE705P
ΝΑ = not applicable NR = not registered
Example 2:
[0071] This example illustrates the production of the legume protein product with reduced astringency of the invention according to the procedure where the acidified legume protein solution has the pH adjusted to precipitate the most astringent protein.
[0072] 18 kg of yellow pea protein concentrate were combined with 300 L of purified water by reverse osmosis (RO) and the mixture stirred for 30 minutes at room temperature. The insoluble material was removed and the sample partially clarified by centrifugation, yielding a protein solution having a protein concentration of 2.47% by weight. To this protein solution were added 51.1 kg of calcium chloride stock solution, prepared by dissolving 8.0 kg of pellets of
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36/72 calcium chloride (95.5%) in 72 L of water. The insoluble material was removed and the sample clarified by centrifugation, yielding 295 L of protein extract solution having a protein concentration of about 1.32% by weight. The 295 L of protein extract solution was combined with 206 L of water per RO and the pH of the sample decreased to 2.75 with HCl solution (concentrated HCl diluted with an equal volume of water). 495 L of acidified protein solution having a protein content of 0.66% by weight were then adjusted to pH 5.5 using 2M NaOH solution, resulting in the formation of a precipitate. 24.92 kg of precipitate were collected by centrifugation yielding 480 L of protein solution from legumes having a protein concentration of 0.20% by weight. The pH of the sample was then adjusted to about 3 with diluted HCl solution and then 480 L of reacidified protein protein solution was concentrated to 28 L using a PES ultrafiltration membrane having a 1000 dalton pore size operated at a temperature of about 58 ° C. 28 L of concentrated protein solution were then diafiltered with 28 L of water per RO at about 63 ° C and still concentrated to provide 19.94 kg of concentrated diafiltered protein solution having a protein content of 6.52% in Weight. This represented a 33.4% yield of the protein in the protein extract solution resulting from the clarification step after the addition of calcium chloride. 19.94 kg of concentrated diafiltered protein solution were spray dried to yield a protein product, having a protein content of 96.07% (N x 6.25) d.b., called YP20-E13-13A YP705.
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[0073] The 24.92 kg of wet precipitate collected, having a protein content of 7.83% by weight, represented a yield of 50.1% of the protein in the protein extract solution resulting from the clarification step after the addition of calcium chloride. A 14.76 kg aliquot of the precipitate was washed with an equal weight of water by RO and then recaptured by centrifugation. This washed precipitate was suspended in fresh water and then spray dried. The dry protein product had a protein content of 95.02% (N x 6.25) d.b. and was named YP20E13-13A YP705P-01. A second aliquot (10 kg) of the precipitate was suspended in water and spray dried without a washing step. The dry protein product had a protein content of 87.52 (N x 6.25) d.b. and was named YP20-E13-13A YP705P-02.
Example 3:
[0074] This example illustrates the production of the legume protein product with reduced astringency of the invention according to the procedure where membrane processing is used to separate the less astringent proteins from the more astringent proteins, 'at' kg of 'b' it was combined with 'c' L of purified water by reverse osmosis (RO) and the mixture stirred for 10 minutes at room temperature. The insoluble material was removed and the sample partially clarified by centrifugation, yielding a protein solution having a protein concentration of 'd'% by weight. To this protein solution was added 'e' g of defoamer and 'f kg of calcium chloride stock solution, prepared by dissolving' g 'kg of calcium chloride pellets (95.5%) in' h 'L in
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38/72 water. The insoluble material was removed and the sample clarified by centrifugation, yielding '1' L of protein extract solution having a protein concentration of 'j'% by weight, 'k' L of protein extract solution was combined with TL of water per RO and the pH of the sample decreased to about 'm' with HCl solution (concentrated HCl diluted with an equal volume of water), 'n' L of acidified protein protein solution, having a protein concentration of The 'wt%' was concentrated to 'p' using a polyvinylidene fluoride (PVDF) microfiltration membrane having a pore size of 0.08 pm operated at a temperature of about 'q' ° C. The microfiltration retentate was then diafiltered with 'r' L of water by RO at about 's' ° C and then the diafiltered retentate further reduced to 't' kg by about 'u' ° C. 'v' L of microfiltration / diafiltration permeate, having a protein concentration of 'w'% by weight, was concentrated to 'x' L using a PES ultrafiltration membrane having a pore size of 1,000 daltons operated at a temperature about 'y' ° C. The concentrated protein solution was then diafiltered with 'z' L of water by RO in about 'aa' ° C 'ab' to provide 'ac' kg of concentrated diafiltered protein solution having a 'ad' protein content % by weight. This represented a yield of 'ae'% of the protein in the protein extract solution resulting from the clarification step after adding calcium chloride, 'af' kg of concentrated diafiltered protein solution was spray-dried to yield a protein product , having a protein content of 'ag'% (N x 6.25) db, called 'ah' 'ai'
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[0075] aj 'aj' kg of retained microfiltration 'ak', having a protein content of 'al'% by weight represented a yield of 'am'% of the protein in the protein extract solution resulting from the clarification step after the addition of calcium chloride, 'an' kg of concentrated microfiltration retentate and diafiltrate was adjusted to pH 'ao' and then spray dried to form a protein product having a protein content of 'ap'% (N x 6.25) db, called 'ah' 'aq'
[007 6] The parameters 'a' to 'ao' are determined in the following table 2.
Table 2 - Parameters for the production of protein products by the membrane fractionation method
ah ΥΡ23Ή12-13Α ΥΡ23Ή14-13Α YP23-J02-13A LE03-D01-14A The 24 24 60 36Concentrate of Concentrate of Concentrate of Flour of B protein from protein from protein from green lentilyellow pea yellow pea yellow pea entire ç 400 400 1008 600 d 3.11 2.92 3.16 1.25 and AT AT 19 AT f 54.6 56.0 135 79.36 g 6 6 20 10 H 54 54 180 90 i 398 398.8 934 604 j 1.66 1.60 About 1.90 0.61 k 398 398.8 934 604 1 269 278.2 666 398 m 3.17 3.16 2.99 3.01 n 670 490 1440 1025
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0 0.86 0.91 0.83 0.30 P 65 L 28.04 kg 180 L 35 L q 59 55 55 56 r AT AT 180 80 s AT AT 55 55 t AT AT 140 AT u AT AT 55 AT V 600 458 About 1470 1052 w 0.18 0.29 0.31 0.27 X 28 30 40 48 y 56 54 56 54 z 140 150 200 96 aa 59 59 58 61 ab And still concentrated AT And still concentrated And still concentrated B.C 21,369 32.35 33.6 32.08 ad 3.43 2.44 5.02 2.09 ae 11.0 12.4 9.5 18.2 af 21.36 32.35 33.6 32.08 ag 101.64 98.24 99.78 93.52 there YP706 YP706 YP706 LE706 aj 65 L 28.04 kg 140 L 32.12 ak Focused Focused Concentrated anddiafiltered Concentrated anddiafiltered al 7.02 9.45 6.63 4.87 am 69.0 41.5 52.3 42.4 an AT AT 135 32.12 to AT AT About 7 7.29 ap AT AT 91.60 94.64
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aq AT AT YP706B LE706B
N / A = not applicable
Example 4:
[0077] This example contains an assessment of dry color and color in solution of protein products from legumes with reduced astringency, produced by the methods of Examples 1-3.
[0078] The color of dry powders was assessed using a HunterLab ColorQuest XE instrument in reflectance mode. The color values are determined in the following table 3:
Table 3 - HunterLab scores for dried legume protein products with reduced astringency
Sample L * The* B* YP20-D23-13AYP705 89.33 0.02 5.75 YP20-D24-13AYP705 88.55 -0.14 5.73 YP20-E02-13AYP705 89.14 0.26 6.68 YP20-E13-13AYP705 86.90 0.90 8.55 LE03-D02-14A LE705 88.09 1.07 5.54 ΥΡ23Ή12-13ΑYP706 88.23 -0.09 6.35 ΥΡ23Ή14-13ΑYP706 88.53 0.22 6.78 YP23-J02-13A YP706 87.25 0.75 7.45 LE03-D01-14A LE706 85.94 0.84 7.92
[0079] As can be seen from Table 3, protein products from legumes with astringency
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42/72 reduced were light in color.
[0080] Solutions of legumes products with reduced astringency were prepared by dissolving enough protein powder to provide 0.48 g of protein in 15 ml of water per RO. The pH of the solutions was measured with a pH meter and the color and clarity were evaluated using a HunterLab Color Quest XE instrument operated in transmission mode. The results are shown in the following table 4.
Table 4 - pH and HunterLab scores for legume protein product solutions with reduced astringency
Sample pH L * The* B* mist YP20-D23-13AYP705 3.35 97.2 -0.10 6.42 22.9 YP20-D24-13AYP705 2.93 97.91 -0.40 5.81 8.2 YP20-E02-13AYP705 3.39 97.76 -0.33 5.52 9.9 YP20-E13-13AYP705 3.26 95.33 0.05 9.69 29.8 LE03-D02-14ALE705 3.21 96.33 0.66 7.18 4.5 ΥΡ23Ή12-13ΑYP706 3.72 94.65 0.01 9.20 14.9 ΥΡ23Ή14-13ΑYP706 3.57 96.07 -0.25 8.99 7.7 YP23-J02-13AYP706 3.51 96.55 0.09 9.7 17.2 LE03-D01-14A 3.42 93.86 0.60 12.8 21.5
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LE706
[0081] As can be seen from the results in
Table 4, the solutions of the legume protein products with reduced astringency were light in color and generally low in fog.
Example 5:
[0082] This example contains an assessment of the water solubility of protein products of legumes with reduced astringency, produced by the methods of Examples 1 and 3. The solubility was tested based on the solubility of the protein (called the protein method, a modified version the procedure by Morr et al., J. Food Sci. 50: 1715-1718) and total product solubility (called the pellet method).
[0083] The protein powder sufficient to provide 0.5 g of protein was weighed in a beaker and moistened by mixing with about 20-25 ml of purified water with reverse osmosis (RO). Additional water was then added to bring the volume to approximately 45 ml. The beaker contents were then slowly stirred for 60 minutes using a magnetic stirrer. The pH was determined immediately after dispersing the protein and was adjusted to the appropriate level (2, 3, 4, 5, 6 or 7) with diluted NaOH or HCl. A sample was also prepared at natural pH. For samples with adjusted pH, the pH was measured and corrected periodically during the 60 minutes of stirring. After 60 minutes of stirring, the samples were prepared in 50 ml of total volume with water by RO, yielding 1% w / v of protein dispersion. The protein content of the dispersions was determined by combustion analysis using a Leco Nitrogen
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Determinator. Aliquots (20 ml) of the dispersions were then transferred to pre-weighed centrifuge tubes, which had been dried overnight in an oven at 100 ° C then cooled in a desiccator and the capped tubes. The samples were centrifuged at 7,800 g for 10 minutes, which sedimented insoluble material and yielded a supernatant. The protein content of the supernatant was measured by combustion analysis and then the supernatant and tube caps were discarded and the pellet material dried overnight in an oven set at 100 ° C. The next morning, the tubes were transferred to a desiccator and allowed to cool. The weight of dry pellet material was recorded. The dry weight of the initial protein powder was calculated by multiplying the weight of the powder used by a factor of ((100 - moisture content of the powder (%)) / 100). The product solubility was then calculated in two different ways:
1) Solubility (protein method) (%) = (% of protein in supernatant /% of protein in initial dispersion) x 100;
2) Solubility (pellet method) (%) = (1 - (insoluble pellet material in dry weight / ((weight of 20 ml weight of dispersion of 50 ml dispersion) x dry protein powder in initial weight)) ) x 100;
3) Values calculated as large as 100% have been reported as 100%.
[0084] The natural pH values of 1% w / v of protein solutions of the protein products produced in examples 1 and 3 are shown in table 5:
Table 5 - Natural pH of pulse solutions with
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Lot Product natural pH YP20-D23-13A YP705 3.36 YP20-D24-13A YP705 3, 15 YP20-E02-13A YP705 3.22 LE03-D02-14A YP705 3, 19 YP23-H12-13A YP706 3.74 YP23-H14-13A YP706 3.53 LE03-D01-14A YP706 3.40
[0085] The obtained solubility results are determined in the following tables 6 and 7:
Table 6 - Solubility of products at different pH values based on the protein method
Solubility (protein method) (%) Lot Product ph 2 ph 3 ph 4 ph 5 ph 6 ph 7 PHNatural YP20-D23-13A YP705 100 100 95.4 94.4 90.1 96.1 98.1 YP20-D24-13A YP705 98.0 100 100 100 93.7 98.1 100 YP20-E02-13A YP705 96.9 100 100 99.0 98.9 93.1 100 LE03-D02-14A YP705 98.0 100 99.1 95.9 100 99.0 96.1 ΥΡ23Ή12-13Α YP706 99.0 100 100 80.2 78.4 92.9 95.2 ΥΡ23Ή14-13Α YP706 100 100 99.0 73.2 77.8 82.7 100 LE03-D01-14A YP706 93.3 100 100 64.6 59.8 64.6 100
Table 7 - Solubility of products at different pH values based on the pellet method
Solubility (protein method) (%) Lot Product ph 2 ph 3 ph 4 ph 5 ph 6 ph 7 natural pH YP20-D23-13A YP705 97.4 98.8 98.4 94.8 92.9 93.7 98.6
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YP20-D24-13A YP705 99.8 100 99.3 98.4 97.4 98.4 99.4 YP20-E02-13A YP705 99.8 99.8 100 96.4 96.9 97.9 99.1 LE03-D02-14A LE705 99.9 100 99.4 94.4 96.3 95.7 99.6 YP23-H12-13A YP706 99.8 99.9 99.1 82.0 79.7 87.8 100 YP23-H14-13A YP706 97.8 97.7 98.5 67.9 81.7 75.0 98.9 LE03-D01-14A LE706 96.8 97.2 96.3 67.1 54.7 68.6 97.4
[0086] As can be seen from the results presented in tables 6 and 7, protein products from legumes with reduced astringency were extremely soluble in the pH range 2-4 and also quite soluble in the pH range 5-7.
Example 6:
[0087] This example contains an assessment of the water clarity of legume protein products with reduced astringency, produced by the methods of Examples 1 and 3.
[0088] The clarity of 1% w / v of protein solutions prepared as described in Example 5 was evaluated by measuring the absorbance at 600 nm (water white), with a lower absorbance score indicating greater clarity. The analysis of the samples in a HunterLab ColorQuest XE instrument in transmission mode also provided a percentage fog reading, another measure of clarity.
[0089] The clarity results are determined in the
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47/72 following tables 8 and 9:
Table 8 - Clarity of protein solutions at different pH values as assessed by A600
A600 Lot Product pH 2 pH 3 pH 4 pH 5 pH 6 pH 7 natural pH YP20-D23-13A YP705 0.0008 0.0016 0.029 0.337 0.807 0.596 0.022 YP20-D24-13A YP705 0.013 0.012 0.021 0.076 0.309 0.213 0.012 YP20-E02-13A YP705 0.007 0.011 0.014 0.063 0.506 0.369 0.012 LE03-D02-14A LE705 0.010 0.012 0.073 0.062 0.027 0.026 0.014 YP23-H12-13A YP706 0.008 0.016 0.034 1,923 1,889 0.791 0.033 YP23-H14-13A YP706 0.011 0.015 0.024 1,931 1,690 1,577 0.018 LE03-D01-14A LE706 0.019 0.025 0.050 2,424 2,412 2,426 0.024
Table 9 - Clarity of protein solutions at different pH values as assessed by HunterLab fog analysis
HunterLab Mist Reading
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Lot Product pH 2 pH 3 pH 4 pH 5 pH 6 pH 7 pHNatural YP20-D23-13A YP705 0.5 5.6 13.0 73.6 90.8 85.3 9.7 YP20-D24-13A YP705 0.0 1.7 6.4 23.1 65.7 50.3 2.2 YP20-E02-13A YP705 0.0 0.8 3.2 16.0 79.5 68.5 1.0 LE03-D02-14A LE705 0.3 1.2 19.8 16.7 3.6 1.8 1.8 ΥΡ23Ή12-13A YP706 0.0 1.0 4.4 96.0 95.8 87.9 4.7 ΥΡ23Ή14-13A YP706 0.0 0.5 2.3 95.9 95.7 95.5 1.1 LE03-D01-14A YP706 3.3 4.9 12.6 100.3 101.3 101.3 4.3
[0090] As can be seen from the results of tables 8 and 9, protein products from legumes with reduced astringency generally provided transparent solutions at pH 2-4.
Example 7:
[0091] This example contains an assessment of the solubility in a soft drink (Sprite) and sports drink (Gatorade of orange) of legumes products with reduced astringency produced by the methods of Examples 1 and 3. Solubility was determined with the protein added to drinks with no pH correction and again with the pH of protein fortified drinks adjusted to the level of the original drinks.
[0092] When solubility has been assessed with no
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49/72 pH correction, a sufficient amount of protein powder to provide 1 g of protein was weighed in a beaker and moistened by mixing with about 20-25 ml of drink. Additional drink was then added to bring the volume to 50 ml and then the solutions were stirred slowly on a magnetic stirrer for 60 minutes to yield a w / v dispersion of 2% protein. The protein content of the samples was determined by combustion analysis using a Leco Nitrogen Determinator then an aliquot of the protein-containing drinks was centrifuged at 7,800 g for 10 minutes and the protein content of the supernatant measured.
[0093] Solubility (%) = (% protein in supernatant /% protein in initial dispersion) x 100.
[0094] Values calculated as more than 100% have been reported as 100%.
[0095] When the solubility was evaluated with pH correction, the pH of the soda (Sprite) and sports drink (Gatorade of orange) without protein was measured. Sufficient amount of protein powder to provide 1 g of protein was weighed in a beaker and moistened by mixing with about 20-25 ml of drink. Additional drink was added to bring the volume to approximately 45 ml, and then the solutions were stirred slowly on a magnetic stirrer for 60 minutes. The pH of protein-containing drinks was determined immediately after dispersing the protein and was adjusted to pH without original protein with HCl or NaOH, as needed. The pH was measured and corrected periodically during the 60 minutes of stirring. After 60 minutes of shaking, the total volume of
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50/72 each solution was placed in 50 ml with additional drink, yielding a w / v dispersion of 2% protein. The protein content of the samples was determined by combustion analysis using a Leco Nitrogen Determinator then an aliquot of the protein-containing drinks was centrifuged at 7,800 g for 10 minutes and the protein content of the supernatant measured.
- Solubility (%) = (% protein in supernatant /% protein in initial dispersion) x 100;
Values calculated as more than 100% have been reported as 100%.
[0096] The results obtained are determined in the following table 10:
Table 10 - Solubility of legume protein products with reduced astringency in Sprite and orange Gatorade
No pH correction pH correction Lot Product Solubility (%)in Sprite Solubility (%) in Gatorade of orange Solubility (%)in Sprite Solubility (%) in Gatorade of orange YP20-D23-13A YP705 100 98.0 97.0 100 YP20-D24-13A YP705 100 97.5 99.5 99.0 YP20-E02-13A YP705 100 100 100 100 LE03- LE705 100 100 98.5 100
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D02-14A YP23-H12-13A YP706 100 99.0 97.0 96.0 YP23-H14-13A YP706 98.5 99.5 98.0 92.1 LE03-D01-14A YP706 92.6 98.9 93.3 100
[0097] As can be seen from the results in Table 10, protein products from legumes with reduced astringency were highly soluble in Sprite and Gatorade orange.
Example 8:
[0098] This example contains an assessment of the clarity in a soft drink and sports drink of protein products from legumes with reduced astringency, produced by the methods of Examples 1 and 3.
[0099] The clarity of the 2% w / v protein dispersion prepared in soft drink (Sprite) and the sports drink (Gatorade of orange) in example 7 were evaluated using the HunterLab fog method described in Example 6.
[0100] The results obtained are determined in the following table 11:
Table 11 - HunterLab fog readings for legume protein products with reduced astringency in Sprite and Orange Gatorade
No pH correction pH correction
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Lot Product Solubility (%)in Sprite Solubility (%) in Gatorade of orange Solubility (%)in Sprite Solubility (%) in Gatorade of orange Without protein0.0 82.6 0.0 82.6 YP20-D23-13A YP705 17.8 70.6 21.8 72.2 YP20-D24-13A YP705 9.4 79.7 12.5 76.3 YP20-E02-13A YP705 8.5 86.2 20.2 86.5 LE03-D02-14A YP705 1.4 85.4 1.7 85.0 YP23-H12-13A YP706 10.2 84.7 6.4 79.9 YP23-H14-13A YP706 4.5 80.6 7.3 78.7 LE03-D01-14A YP706 11.5 77.5 12.1 78.9
[0101] As can be seen from the results of
Table 11, the addition of protein products from legumes with reduced astringency to soda and sports drink added little or no cloudiness.
Example 9:
[0102] This example contains an assessment of the thermal stability in water of protein products of legumes with reduced astringency, produced by the methods of Examples 1 and 3.
[0103] protein solutions w / v to 2% of the protein products were prepared in water by RO. The pH of the solutions
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53/72 was determined with a pH meter and then adjusted to about 3.0 with HCl solution. The clarity of the solutions was assessed by measuring the mist with the HunterLab Color Quest XE instrument operated in transmission mode. The solutions were then heated to 95 ° C, maintained at this temperature for 30 seconds and then immediately cooled to room temperature in an ice bath. The clarity of the heat-treated solutions was then measured again.
[0104] The clarity of the protein solutions before after heating is determined in Table 12 below:
Table 12 - Effect of heat treatment on the clarity of 2% w / v protein solutions of legumes products with reduced astringency
Lot Product Mist before heat treatment(%) Mist after theheat treatment(%) YP20-D23-13A YP705 13.0 0.0 YP20-D24-13A YP705 4.2 0.0 YP20-E02-13A YP705 5.5 1.4 LE03-D02-14A YP705 1.0 0.0 ΥΡ23Ή12-13Α YP706 5.0 2.0 ΥΡ23Ή14-13Α YP706 3.3 2.2 LE03-D01-14A LE706 6.3 1.6
[0105] As can be seen from the results in Table 13, the protein products solutions of legumes with reduced astringency were substantially clear before heat treatment and the mist level was currently reduced by heat treatment.
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Example 10:
[0106] This example illustrates the production of protein products from legumes by the method described in United States Patent Application 13 / 556,357.
[0107] 'a' kg of 'b' was combined with 'c' L of 'd' in 'e' and stirred for 'f' minutes, 'g' kg of calcium chloride pellets (95.5%) dissolved in 'h' L of water per RO was then added and the mixture stirred for an additional '1' of minutes. Residual solids were removed by centrifugation to produce a centrifugate having a protein content of 'j'% by weight, 'k' L of centrifugate was added to '1' L of water by RO in 'm' and the pH of the sample decreased to 'n' with diluted HCl. The diluted and acidified centrifugate was further clarified by filtration to provide a clear protein solution with a protein content of 'o'% by weight.
[0108] The filtered protein solution was reduced in volume from 'p' L to 'q' L by concentration on a polyethersulfone membrane, having a molecular weight cut of 'r' daltons, operated at a temperature of around ' s' ° C. At this point, the protein solution, with a protein content of 't'% by weight, was diafiltered with 'u' L of water by RO, with the diafiltration operation conducted at about 'ν' ° C. The diafiltered protein solution was then further concentrated in 'w' kg, having a protein content of 'x'% by weight, then diluted with water by RO to a protein content of 'y'% by weight to facilitate spray drying. The protein solution before spray drying, having a weight of 'z' kg was recovered in a yield of 'aa'% of the initial centrifuge which was still
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55/72 processed. The concentrated and diafiltered protein solution was then dried to yield a product which was found to have a protein content of 'ab'% by weight (N x 6.25) d.b .. The product received the desognation 'ac'.
[0109] The parameters 'a' to 'ac' are determined in the following table 13.
Table 13 - Parameters for executions to produce 701 wrist products
B.C YP01-E19-11AYP701 YP05-E18-12AYP701 LE01-J24-13A LE701 The 20 70 20 B pea flouryellow match pea flouryellow match lentil flourwhole green ç 200 300 200 d 0.15 M CaCI ₂ RO water 0.13 M CaCI ₂ and 60 ^S C 30 ^S C Temperatureenvironment f 30 60 30 g 0 4.52 0 H 0 10 0 i 0 30 0 j 1.32 2.92 1.65 k 186.5 223.3 146.2 1 225.8 223.0 147.7 m 60 ^S C Temperatureenvironment Temperatureenvironment n 3.34 3.04 2.65 0 0.58 1.25 0.62 P 400 550 295 q 35 101 25
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r 100,000 10,000 100,000 s 58 53 30 t 4.94 4.05 4.23 u 350 202 250 V 60 53 32 w 21.52 34.78 21.60 X 7.54 10.02 4.69 y AT 5.00 AT z 21.52 57.90 21.60 aa 65.9 44.5 41.9 ab 103.19 101.99 103.11
N / A = not applicable
Example 11:
[0110] This example illustrates a comparison of the astringency level of the YP20-D24-13A YP705 prepared as described in Example 1 with that of YP01-E19-11A YP701 prepared as described in example 10.
[0111] Samples were prepared for sensory evaluation by dissolving enough protein powder to provide 5 g of protein in 250 ml of purified drinking water. The initial pH of the YP705 solution was 3.09 and it was adjusted to about 3.50 with food-grade sodium hydroxide solution. The initial pH of the YP701 solution was 3.92 and it was adjusted to about 3.50 with food grade hydrochloric acid. An informal panel of seven respondents was asked to blindly taste the samples and indicate which was less astringent.
[0112] Five out of seven respondents indicated that the YP20-D24-13A YP705 was less astringent.
Example 12:
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[0113] This example illustrates a comparison of the astringency level of YP20-E02-13A YP705 prepared as described in example 1 with that of YP01-E19-1 ΙΑ YP701 prepared as described in example 10.
[0114] Samples were prepared for sensory evaluation by dissolving enough protein powder to provide 5 g of protein in 250 ml of purified drinking water. The initial pH of the YP705 solution was 3.38 and it was adjusted to about 3.50 with food grade sodium hydroxide solution. The initial pH of the YP701 solution was 3.94 and it was adjusted to about 3.50 with food grade hydrochloric acid. An informal panel of seven respondents was asked to taste the samples blindly and indicate which were less astringent.
[0115] Five out of seven respondents indicated that the YP20-E02-13A YP705 was less astringent.
Example 13:
[0116] This example illustrates a comparison of the astringency level of YP20-E13-13A YP705 prepared as described in example 2 with that of YP05-A18-12A YP701 prepared as described in example 10.
[0117] Samples were prepared for sensory evaluation by dissolving enough protein powder to provide 5 g of protein in 250 ml of purified drinking water. The two samples had pH values within 0.1 units of each other so that no pH adjustment was made. An informal panel of eight respondents was asked to blindly taste the samples and indicate which ones were less astringent. The experiment was conducted a second time with a panel of ten members. The results
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Cumulative 58/72 are presented below.
[0118] Eleven out of eighteen respondents indicated that the YP20-E13-13A YP705 was less astringent.
Example 14:
[0119] This example illustrates a comparison of the astringency level of YP20-H12-13A YP706 prepared as described in example 1 with that of YP05-A18-12A YP701 prepared as described in example 10.
[0120] Samples were prepared for sensory evaluation by dissolving enough protein powder to provide 5 g of protein in 250 ml of purified drinking water. The initial pH of the YP706 solution was 3.72 and it was adjusted to about 3.50 food grade hydrochloric acid. The initial pH of the YP701 solution was 3.17 and it was adjusted to about 3.50 with food grade sodium hydroxide solution. An informal panel of seven respondents was asked to taste the samples blindly and indicate which were less astringent.
[0121] Four out of seven respondents indicated that the YP20-H12-13A YP706 was less astringent.
Example 15:
[0122] This example illustrates a comparison of the astringency level of YP20-H14-13A YP706 prepared as described in example 1 with that of YP05-A18-12A YP701 prepared as described in example 10.
[0123] Samples were prepared for sensory evaluation by dissolving enough protein powder to provide 5 g of protein in 250 ml of purified drinking water. The initial pH of the YP701 solution was 3.12 and it was adjusted to 3.48 with sodium hydroxide solution with
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59/72 food. The pH of the YP706 solution was 3.46. An informal panel of seven respondents was asked to taste the samples blindly and indicate which were less astringent.
[0124] Five out of seven respondents indicated that the YP20-H14-13A YP706 was less astringent.
Example 16:
[0125] This example illustrates a comparison of the astringency level of LE03-D02-14A LE705 prepared as described in example 1 with that of LE01-J24-13A YP701 prepared as described in example 10.
[0126] Samples were prepared for sensory evaluation by dissolving enough protein powder to provide 5 g of protein in 250 ml of purified drinking water. The initial pH of the LE705 solution was 3.17 and it was adjusted to 3.47 with food-grade sodium hydroxide solution. The initial pH of the LE701 solution was 3.81 and it was adjusted to 3.52 with food grade hydrochloric acid. An informal panel of eight respondents was asked to blindly taste the samples and indicate which ones were less astringent.
[0127] Six out of eight respondents indicated that LE03D02-14A LE705 was less astringent.
Example 17:
[0128] This example illustrates a comparison of the astringency level of LE03-D01-14A LE706 prepared as described in example 3 with that of LE01-J24-13A LE701 prepared as described in example 10.
[0129] Samples were prepared for sensory evaluation by dissolving enough protein powder to
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60/72 provide 5 g of protein in 250 ml of purified water for drinking. The initial pH of the LE706 solution was 3.37 and it was adjusted to about 3.5 with food-grade sodium hydroxide solution. The pH of the LE701 solution was 3.84 and it was adjusted to about 3.5 with a food grade hydrochloric acid solution. An informal panel of eight respondents was asked to blindly taste the samples and indicate which ones were less astringent.
[0130] Five out of eight respondents indicated that LE03-D01-14A LE706 was less astringent.
Example 18:
[0131] This example contains an assessment of the dry color and color in solution of the by-products of the production of legume protein products with reduced astringency, prepared according to the methods of Examples 1-3.
[0132] The color of dry powders was assessed using a HunterLab ColorQuest XE instrument in reflectance mode. The color values are determined in the following table 14:
Table 14 - HunterLab scores for dry protein products
Sample L * The* B* YP20-D23-13A YP705P 84.78 1.30 9.87 YP20-D24-13A YP705P 88.97 0.21 6.08 YP20-E02-13A YP705P 89.06 0.22 6.37 YP20-E13-13A YP705P-01 82.64 1.99 12.53 YP20-E13-13A YP705P-02 83.61 1.80 11.06 LE03-D02-14A LE705P 74.27 1.53 8.32 YP23-J02-13A YP706B 81.57 1.32 10.45 LE03-D01-14A LE706B 78.19 1.96 8.35
[0133] As can be seen from the results in
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Table 14, by-products were generally darker, redder and more yellow than protein products from legumes with reduced astringency.
[0134] Solutions of the co-products of the preparation of legume protein products with reduced astringency were prepared by dissolving enough protein powder to provide 0.48 g of protein in 15 ml of water per RO. The pH of the solutions was measured with a pH meter and the color and clarity were evaluated using a HunterLab Color Quest XE instrument operated in transmission mode. The results are shown in the following table 15.
Table 15 - pH and HunterLab scores for legume protein product solutions
Sample pH L * The* B* mist YP20-D23-13AYP705P 5.81 43.87 5.5 28.43 97.1 YP20-D24-13AYP705P 6.13 40.94 6.82 30.44 97.3 YP20-E02-13AYP705P 4.95 39.68 6.79 31.08 99.2 YP20-E13-13AYP705P-01 5.29 39.32 8.4 33.01 96.5 YP20-E13-13AYP705P-02 5.03 32.10 10.7 34.12 96.4 LE03-D02-14ALE705P 6.40 11.69 11.81 17.59 97.9 YP23-J02-13AYP706B 7.39 41.26 7.88 31.65 95.7 LE03-D01-14ALE706B 7.09 38.09 7.75 25.18 97.3
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[0135] As can be seen from the results in Table 15, the solutions of the co-products were all with a very high mist content. The solutions were also darker, redder and more yellow than the solutions of the wrist products with reduced astringency.
Example 19:
[0136] This example contains an assessment of the water solubility of the byproducts of the production of wrist products with reduced astringency, prepared by the methods of Examples 1 and 3. The solubility was tested based on the solubility of the protein (called the protein method, a modified version of the procedure by Morr et al., J. Food Sci. 50: 1715-1718).
[0137] Sufficient protein powder to provide 0.5 g of protein was weighed in a beaker and moistened by mixing with about 20-25 ml of purified water with reverse osmosis (RO). Additional water was then added to bring the volume to approximately 45 ml. The beaker contents were then slowly stirred for 60 minutes using a magnetic stirrer. The pH was determined immediately after dispersing the protein and was adjusted to the appropriate level (6, 6.5, 7, 7.5 or 8) with diluted NaOH or HCl. The pH was then measured and corrected periodically during the 60 minutes of stirring. After 60 minutes of stirring, the samples were constituted in 50 ml of the total volume with water by RO, yielding a dispersion of protein at 1% w / v. The protein content of the dispersions was determined by combustion analysis using a Leco Nitrogen Determinator. The samples were then centrifuged at 7,800 g for 10 minutes, which sedimented the insoluble material and
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63/72 yielded a supernatant. The protein content of the supernatant was measured by combustion analysis.
[0138] Solubility of product was Next calculated: 1) Solubility (method in protein) (%) (% in supernatant protein /% in protein in dispersal initial) x 100; 2) Values calculated as more than what 100% were
reported as 100%.
[0139] The obtained solubility results are determined in the following table 16:
Table 16- Solubility of products at different pH values based on the protein method
Solubility (protein method) (%) Lot Product pH 6 pH 6.5 pH 7 pH 7.5 pH 8 YP20-D23-13A YP705P 5.7 2.9 9.9 12.0 11.8 YP20-D24-13A YP705P 13.0 9.9 15.2 11.7 15.3 LE03-D02-14A LE705P 13.6 10.9 11.0 11.7 9.6 YP23-J02-13A YP706B 16.5 15.5 20.4 17.7 9.6 LE03-D01-14A LE706B 2.0 1.8 4.7 9.3 5.1
[0140] As can be seen from the results in Table 16, the co-products of the production of protein products from legumes with reduced astringency were poorly soluble over a pH range of 2 to 7.
Example 20:
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[0141] This example contains an assessment of the water-binding capacity of the by-products of the production of wrist products with reduced astringency, prepared by the methods of Examples 1 and 3.
[0142] Protein powder (1 g) was weighed in centrifuge tubes (50 ml) of known weight. To this powder approximately 20 ml of purified water by reverse osmosis (RO) at natural pH were added. The contents of the tubes were mixed using a vortex mixer at moderate speed for 1 minute. The samples were incubated at room temperature for 5 minutes thereafter, mixed with the vortex mixer for 30 seconds. This was followed by incubation at room temperature for another 5 minutes followed by another 30 seconds of vortex mixing. The samples were then centrifuged at 1,000 g for 15 minutes at 20 ° C. After centrifugation, the supernatant was carefully poured, ensuring that all solid material remains in the tube. The centrifuge tube was then re-weighed and the weight of the sample saturated with water was determined.
[0143] The water binding capacity (WBC) w was calculated as:
WBC (ml / g) = (sample mass saturated with water initial sample mass) / (initial sample mass x total sample solids content)
[0144] The results of the water binding capacity obtained are determined in the following table 17.
Table 17 - Water binding capacity of various products
Product WBC (ml / g)
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YP20-D23-13A YP705P 2.60 YP20-D24-13A YP705P 2.59 LE03-D02-14A LE705P 3.90 YP23-J02-13A YP706B 2.88 LE03-D01-14A LE706B 2.74
[0145] As can be seen from the results in Table 17, all byproducts from the production of legume protein products with reduced astringency had moderate water binding capacities.
Example 21:
[0146] This example illustrates the preparation of a legume protein isolate by conventional isoelectric precipitation.
[0147] 20 kg of yellow pea protein concentrate was added to 200 L of water per RO at room temperature and the pH adjusted to about 8.5 by the addition of sodium hydroxide solution. The sample was stirred for 30 minutes to provide an aqueous protein solution. The extraction pH was monitored and maintained at about 8.5 over the 30 minutes. The residual pea protein concentrate was removed and the resulting protein solution clarified by centrifugation and filtration to produce 240 L of filtered protein solution having a protein content of 3.52% by weight. The pH of the protein solution was adjusted to about 4.5 by the addition of HCl which had been diluted with an equal volume of water and a precipitate was formed. The precipitate was collected by centrifugation, then washed by resuspending it in 2 volumes of water by RO. The washed precipitate was then collected by centrifugation. A total of 30.68 kg of
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66/72 washed precipitate was obtained with a protein content of 22.55% by weight. This represented an 81.9% yield of the protein in the clarified extract solution. A 15.34 kg aliquot of the washed precipitate was combined with 15.4 kg of water per RO and then the pH of the sample adjusted to about 7 with sodium hydroxide solution. The pH-adjusted sample was then spray-dried to yield an isolate with a protein content of 90.22% (N x 6.25) d.b .. The product was designated conventional YP12-K13-12A IEP pH 7.
Example 22:
[0148] This Example is a sensory evaluation of the product YP20-D23-13A YP705P prepared as described in example 1 with the product of pea protein isolate prepared as described in example 21.
[0149] Samples were prepared for sensory evaluation by dissolving enough protein powder to provide 5 g of protein in 250 ml of purified drinking water. The initial pH of the conventional YP12-K13-12A IEP pH 7 solution was 7.08. The initial pH of the YP705P solution was 5.77 and it was adjusted to 7.08 with a food-grade sodium hydroxide solution. An informal panel of eight respondents was asked to blindly taste the samples and indicate which ones tasted cleaner and which sample they preferred.
[0150] Seven out of eight respondents preferred the YP20D23-13A YP705P and seven out of eight considered it to have a cleaner flavor.
Example 23:
[0151] This Example is a sensory evaluation of the product YP20-D24-13A YP705P prepared as described in
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67/72 example 1 with the conventional pea protein isolate product prepared as described in example 21.
[0152] Samples were prepared for sensory evaluation by dissolving enough protein powder to provide 5 g of protein in 250 ml of purified drinking water. The initial pH of the conventional YP12-K13-12A IEP pH 7 solution was 7.06. The initial pH of the YP705P solution was 6.18 and it was adjusted to 7.10 with food-grade sodium hydroxide solution. An informal panel of nine respondents was asked to blindly taste the samples and indicate which ones tasted cleaner and which sample they preferred.
[0153] All nine respondents preferred the YP20D24-13A YP705P and considered it to have a cleaner taste.
Example 24:
[0154] This Example is a sensory evaluation of the product of YP23-J02-13A YP706B prepared as described in example 3 with the product of conventional pea protein isolate prepared as described in example 21.
[0155] Samples were prepared for sensory evaluation by dissolving enough protein powder to provide 5 g of protein in 250 ml of purified drinking water. The pH of the YP12-K13-12A IEP pH 7 solution was 7.09. The pH of the YP23-J02-13A YP706B solution was adjusted to 7.04 with food grade hydrochloric acid. An informal panel of eight respondents was asked to blindly taste the samples and indicate which tasted cleaner and which sample they preferred. The experiment was conducted a second time with a panel having 7 members. Cumulative results are shown below.
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[0156] Eleven out of fifteen respondents considered the YP23-J02-13A YP706B to have the cleanest taste. Ten out of fifteen respondents preferred the YP23-J02-13A YP706B.
Example 25:
[0157] This example illustrates the molecular weight profile of legume protein products prepared as described in examples 1-3 as well as the molecular weight profile of some commercial yellow pea protein products (Propulse (Nutri-Pea, Portage la Prairie, MB), Nutralys S85F (Roquette America, Inc. Keokuk, LA) and Pisane C9 (Cosucra Groupe Warcoing SA, Belgium) These protein products were chosen because they were among the most highly purified pea protein ingredients today commercially available.
[0158] Molecular weight profiles were determined by size exclusion chromatography using a Varian ProStar HPLC system equipped with a 300 x 7, 8 mm Phenomenex BioSep series S-2000 column. The column contained rigid support means of hydrophilic bound silica, 5 micron diameter, pore size of 145 Angstrom.
[0159] before the legume protein samples were analyzed, a standard curve was prepared using a Biorad protein standard (Biorad product No. 151-1901) containing proteins with known molecular weights between 17,000 Daltons (myoglobin) and 670,000 Daltons (thyroglobulin ) with Vitamin B 12 added as a low molecular weight marker in 1,350 Daltons. A standard 0.9% w / v protein solution was prepared in water, filtered with a 0.45 pm pore size filter disc following an aliquot 50 run on
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69/72 column using a mobile phase of 0.05M phosphate / 0.15M NaCl, pH 6 containing 0.02% sodium azide. The flow rate of the mobile phase was 1 mL / min and the components were detected based on the absorbance at 280 nm. Based on the retention times of these molecules of known molecular weight, a regression formula was developed related to the natural log of the molecular weight to the retention time in minutes.
[0160] Retention time (min) = -0.955 x Em (molecular weight) + 18.502 (r2 = 0.98).
[0161] For the analysis of legume protein samples, 0.05 M NaCl, pH 3.5 containing 0.02% sodium azide was used as the mobile phase and also to dissolve dry samples. The protein samples were mixed with the mobile phase solution at a concentration of 1% w / v, placed on a shaker for at least 1 hour then filtered using 0.45 pm pore size filter discs. The sample injection size was 50 pL. The flow rate of the mobile phase was 1 mL / minute and the components were detected based on the absorbance at 280 nm.
[0162] The regression formula above relating molecular weight and retention time, was used to calculate the retention times that correspond to the molecular weights of 100,000 Da, 15,000 Da, 5,000 Da and 1,000 Da. The ProStar HPLC system was used to calculate the peak areas that are within these retention time ranges and the percentage of protein ((peak area of the range / peak area of the total protein) x 100) falling within a given molecular weight range was calculated. Note that the data was not corrected due to the protein response.
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[0163] The molecular weight profiles of the products prepared as described in examples 1-3 and the commercial products are shown in table 18.
Table 18 - Molecular Weight Profile of Products
Pulse Protein
Product %> 100.00 Da % 15,000-100,000 Da % 5,000 -15,000 Da % 1,000 - 5,000Gives YP20-D23-13AYP705 31 33 31 5 YP20-D24-13AYP705 30 36 29 5 YP20-E02-13AYP705 31 37 58 4 YP20-E13-13AYP705 66 16 14 4 LE03-D02-14ALE705P 37 38 16 9 ΥΡ23Ή12-13ΑYP706 21 38 16 9 ΥΡ23Ή14-13ΑYP706 28 29 36 7 YP23-J02-13AYP706 16 28 48 8 LE03-D01-14ALE706 39 24 18 9 YP20-D23-13AYP705P 22 29 34 15 YP20-D24-13AYP705P 21 30 33 17 YP20-E02-13A 24 32 30 15
Petition 870170036348, of 05/30/2017, p. 74/86
71/72
YP705P YP20-E13-13AYP705P-01 27 26 19 29 YP20-E13-13AYP705P-02 38 22 17 24 LE03-D02-14ALE705P 35 37 22 6 YP23-J02-13AYP706B 38 28 14 20 LE03-D01-14ALE706B 75 16 3 5 Nutralys S85f 7 29 9 56 Pisane C9 5 31 29 36 Propulse 13 25 18 45
[0164] As can be seen from the results shown in Table 18, the molecular weight profiles of the products prepared according to Examples 1-3 were different from the molecular weight profiles of commercial yellow pea protein products.
Example 26:
[0165] This example contains an assessment of the phytic acid content of the legume protein products produced as described in examples 1 to 3. The phytic acid content was determined using the method of Latta and Eskin (J. Agric. Food Chem. , 28: 1313-1315).
[0166] The results obtained are determined in the following table 19.
Table 19 - Phytic acid content of protein products
Product % phytic acid d.b. YP20-D23-13A YP705 0.00
Petition 870170036348, of 05/30/2017, p. 75/86
72/72
YP20-D24-13A YP705 0.00 YP20-E02-13A YP705 0.02 YP20-E13-13A YP705 0.00 LE03-D02-14A LE705 0.19 P23Ή12-13Α YP706 0.00 P23Ή14-13Α YP706 0.00 YP23-J02-13A YP706 0.01 LE03-D02-14A LE705 0.29 YP20-D23-13A YP705P 0.02 YP20-D24-13A YP705P 0.01 YP20-E02-13A YP705P 0.06 YP20-E13-13A YP705P-01 0.00 YP20-E13-13A YP705P-02 0.00 LE03-D02-14A LE705P 0.23 YP23-J02-13A YP706B 0.10 LE03-D01-14A LE706B 0.21
[0167] As can be seen from the results in Table 19, all products tested were low in phytic acid.
Description Summary
[0168] In the summary of this description, the present invention provides legume protein products, preferably isolated from legume protein, which have reduced astringency when savored in an acidic solution such as an acidic drink. Modifications are possible within the scope of this invention.

权利要求:
Claims (19)
[1]
1. Method of preparing the protein product of legumes with reduced astringency when tested in aqueous solution at a pH below 5, CHARACTERIZED by the fact that it comprises:
(a) extracting a legume protein source with an aqueous solution of calcium salt to cause the legume protein to solubilize from the protein source and to form an aqueous legume protein solution;
(b) separating the aqueous legume protein solution from the residual legume protein source;
(c) optionally diluting the aqueous protein solution of legumes;
(d) adjusting the pH of the aqueous vegetable protein solution to a pH of 1.5 to 4.4 to produce an acidified vegetable protein solution;
(e) optionally clarifying the acidified legume protein solution if it is not yet clear;
(f) alternatively from steps (b) to (e), optionally dilute and then adjust the pH of the combined aqueous legume protein solution and the residual legume protein source to a pH of 1.5 to 4.4 and then separating the acidified legume protein solution, preferably clear, from the residual legume protein source; and (g) fractionating the proteins in the acidified legume protein solution to separate the lower molecular weight proteins, less astringent from the higher molecular weight proteins, but astringent.
[2]
2. Method, according to claim 1, CHARACTERIZED
Petition 870170036348, of 05/30/2017, p. 77/86 due to the fact that the said fractioning step is carried out by:
(i) adjusting the pH of the acidified legume protein solution to a pH value of 5 to 6.5 to precipitate higher molecular weight proteins, more astringent from the acidified legume protein solution and providing a legume protein solution adjusted to pH;
(ii) removing the precipitate from the pH-adjusted protein protein solution;
(iii) adjusting the pH of the pH-adjusted protein protein solution to a pH value of 1.5 to 4.4, to form a reacidified aqueous protein protein solution; and (iv) drying the reacidified aqueous soy protein solution to provide a less astringent legume protein product.
[3]
3. Method, according to claim 2, CHARACTERIZED
by the fact that stage ( i) from adjustment of pH is carried out in a pH of 5.5 to 6.0.4. Method, of wake up with claim 2 or 3, CHARACTERIZED BY fact of that stage ( (iii ) of the pH is carried out at a pH from 2 to 4.
[4]
5. Method according to any of the claims
1 to 4, CHARACTERIZED by the fact that the aqueous protein solution of leguminized protein from step 1 (e) or (f) as defined in claim 1 is concentrated before step (i) as defined in claim 2 or the protein solution of reacidified legumes from step (iii) as defined in claim 2 is concentrated to increase the protein concentration from 50 to 300 g / L, or from 100 to 200 g L, or
Petition 870170036348, of 05/30/2017, p. 78/86 partially concentrated before step (i) as defined in claim 2 for a protein concentration in the case of the acidified aqueous protein protein solution of less than 50 g / L and in the case of the reacidified protein protein solution less than 10 g / L, while keeping its ionic resistance substantially constant.
[5]
6. Method, according to claim 5, CHARACTERIZED by the fact that said concentration step is carried out using ultrafiltration using a membrane with a molecular weight cut of 1,000 to 1,000,000 daltons, or 1,000 to 100,000 daltons, or 1,000 to 10,000 daltons.
[6]
7. Method according to claim 5 or 6, CHARACTERIZED by the fact that the concentrated or partially concentrated acidified protein protein solution or concentrated or partially concentrated reacidified protein solution or acidified protein solution before concentration or the leguminous protein solution reacidified before concentration is diafiltered, and in the case of diafiltration of the solution before concentration or diafiltration of the partially concentrated solution, the diafiltered solution is concentrated to a concentration of 50 to 300 g / L, or 100 to 200 g / L for acidified legume protein solution and a concentration of 10 to 300 g / L, or 100 to 200 g / L for reacidified legume protein solution.
[7]
8. Method, according to claim 7, CHARACTERIZED by the fact that the diafiltration step is carried out in the optional presence of an antioxidant such as sodium sulfite or ascorbic acid, in the amount of 0.01 to 0.1% by weight , or 0.05% by weight, using 1 to 40 volumes, or
Petition 870170036348, of 05/30/2017, p. 79/86
2 to 25 volumes of diafiltration solution, using a membrane that has a molecular weight cut of 1,000 to 1,000,000 daltons, or 1,000 to 100,000 daltons, or 1,000 to 10,000 daltons.
[8]
9. Method, according to claim 8, CHARACTERIZED by the fact that the diafiltration operation is carried out until no significant additional amounts of contaminants or visible color are present in the permeate, or in the case of the reacidified protein solution, until the retentate has been sufficiently purified to provide, when dried, a legume protein isolate that has a protein content of at least 90% by weight (N x 6.25) db.
[9]
10. Method, according to claim 2, CHARACTERIZED by the fact that the precipitate removed from step (ii), is further processed by a step selected from the group consisting of:
(i) optionally washing the removed precipitate and drying the washed precipitate;
(ii) optionally washing the removed precipitate, adjusting the pH of the precipitate from 6 to 8 and drying the precipitate with adjusted pH;
(iii) adjusting the pH of the removed precipitate to 1.5 to 4.4, or to 2 to 4, processing the membrane to remove contaminants and drying the processed precipitate from the membrane; and (iv) adjust the pH of the removed precipitate to 1.5 to 4.4, or to 2 to 4, process the membrane to remove contaminants, adjust the pH of the processed membrane solution to 6 to 8, and dry the solution with adjusted pH.
[10]
11. Method according to claim 1,
Petition 870170036348, of 05/30/2017, p. 80/86
CHARACTERIZED by the fact that the said fractionation step is carried out by:
(i) processing the membrane of the acidified aqueous protein protein solution to fractionate the protein components of the acidified aqueous protein protein solution into a fraction of higher molecular weight in a first retentate and a fraction of lower molecular weight in a first permeate ;
(ii) processing the membrane of the first permeate to retain protein components of lower molecular weight fraction in a second retentate and to allow contaminants to pass through the membrane in a second permeate; and (iii) drying the second retentate to provide a reduced astringency protein protein product.
[11]
12. Method according to claim 11, CHARACTERIZED by the fact that the membrane processing step (i) is carried out by microfiltration using membranes that have a pore size of 0.05 to 0.1 pm, or 0 , 08 to 0.1 pm or ultrafiltration using membrane with a molecular weight cut of 10,000 to 1,000,000 daltons, or 100,000 to 1,000,000 daltons to concentrate the acidified aqueous protein protein solution at a protein concentration of 50 at 300 g / L, or from 100 to 200 g / L, to provide a concentrated retentate.
[12]
13. Method, according to claim 12, CHARACTERIZED by the fact that the concentrated retentate is subjected to a diafiltration step using from 1 to 40 volumes of diafiltration solution, or from 2 to 25 volumes of diafiltration solution.
Petition 870170036348, of 05/30/2017, p. 81/86
[13]
14. Method according to any one of the claims
11 to 13, CHARACTERIZED by the fact that the processing of the membrane of the first permeate in step (ii) is carried out by ultrafiltration to concentrate the first permeate at a concentration of 10 to 300 g / L, or from 100 to 200 g / L, followed by optional diafiltration, or at a partial concentration of less than 10 g L, using membranes that have a molecular weight cut of 1,000 to 100,000 daltons, or 1,000 to 10,000 daltons.
[14]
15. Method, according to any of claims 11 to 13, CHARACTERIZED by the fact that the first retentate of step (i) is further processed by a step selected from the group consisting of:
(i) drying the first retentate;
(ii) adjust the pH of the first retentate to a pH of 6 to
8, and dry the pH-adjusted retentate.
[15]
16. Legume protein product (A) which has a protein content of at least 60% by weight, or 90% by weight or 100% by weight (N x 6.25) d.b., FEATURED by the fact that:
- it is completely soluble in aqueous media at acid pH values of less than 4.4;
- is heat-stable in aqueous media at acid values of less than 4.4;
- does not require stabilizers or other additives to keep the protein product in solution or suspension;
- is low in phytic acid;
- does not require enzymes in its production;
- it is low in astringency when tested in the aqueous solution at a pH below 5; or
Petition 870170036348, of 05/30/2017, p. 82/86 a protein product from legumes (B) which has a protein content of at least 60% by weight, or 90% by weight or 100% by weight (N x 6.25) d.b. and which has low astringency when tested in the aqueous solution at a pH below 5 which is substantially completely soluble in an aqueous medium at a pH of less than 4.4; or a legume protein product (C), which has a protein content of at least 60% by weight, or 90% by weight or 100% by weight (N x 6.25) db, which has a solubility in water of 1 % w / v protein at a pH of 2 to 7 of more than 50%, as determined by the methods defined in Example 5; or a legume protein product (D that has a molecular weight profile, as determined by the methods defined in Example 25, which is:
10 to 75% greater than 100,000 Da or 15 to 40% greater than 100,000 Da,
10 to 45% of 15,000 to 100,000 Da or 15 to 40% greater than
100,000 Da,
8 to 55% of 5,000 to 15,000 Da or 15 to 50% of 5,000 to
15,000 Da,
2 to 12% from 1,000 to 5,000 Da or 3 to 10% from 1,000 to 5,000 Da; or a Legume protein product (E), which has a molecular weight profile, as determined by the methods defined in Example 25, which is:
10 to 85% greater than 100,000 Da or 18 to 78% greater than 100,000 Da,
10 to 45% from 15,000 to 100,000 Da or 15 to 38% from 15,000 to 100,000 Da,
Petition 870170036348, of 05/30/2017, p. 83/86
0 to 40% from 5,000 to 15,000 Da or 2 to 35% from 5,000 to 15,000
Gives,
1 to 34% from 1,000 to 5,000 Da or 3 to 25% from 1,000 to 5,000 Da.
[16]
17. Legume protein product according to claim 16, CHARACTERIZED by the fact that the legume protein has not been hydrolyzed.
[17]
18. Legume protein product according to claim 16 or 17, characterized in that it has a phytic acid content of less than 1.5% by weight or less than 0.5% by weight.
[18]
19. Legume protein product according to any one of claims 16 to 18, CHARACTERIZED by the fact that it is mixed with water-soluble powder materials for the production of aqueous solutions of the mixture.
[19]
20. Aqueous solution of the legume protein product as defined in any of claims 16 to 18, CHARACTERIZED by the fact that it is heat-stable at a pH of less than 4.4 and that it is a drink that is a clear beverage in which the dissolved legume protein product is completely soluble and transparent or is a non-transparent drink in which the dissolved legume protein increases or does not cloud or cloud.

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引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

US5844086A|1996-01-31|1998-12-01|Stilts Corporation|Oil seed protein extraction|

---

GB0329833D0|2003-12-23|2004-01-28|Unilever Plc|Beverages and their preparation|

---

NZ616202A|2008-10-21|2015-04-24|Burcon Nutrascience Mb Corp|Production of soluble protein solutions from soy |

---

CN102387713B|2009-02-11|2015-07-29|伯康营养科学公司|Soy protein products is produced with calcium chloride extraction|

---

US8404299B2|2009-06-30|2013-03-26|Burcon Nutrascience Corp.|Preparation of soy protein isolate using calcium chloride extraction |

---

PT2482670T|2009-06-30|2017-12-11|Burcon Nutrascience Corp|Preparation of soy protein isolate using calcium chloride extraction |

---

US8128993B2|2009-07-31|2012-03-06|Nantero Inc.|Anisotropic nanotube fabric layers and films and methods of forming same|

---

US10506821B2|2010-05-07|2019-12-17|Burcon Mutrascience Corp.|Production of soluble protein solutions from pulses|

---

US20120135117A1|2010-05-07|2012-05-31|Segall Kevin I|Production of soluble protein solutions from pulses|

---

KR20130079408A|2010-05-07|2013-07-10|버콘 뉴트라사이언스 코포레이션|A method of producing soluble protein solutions from pulses and product thereof|

---

WO2012083418A1|2010-11-24|2012-06-28|Burcon Nutrascience Corp.|Astringency in soy protein solutions|

---

US9635875B2|2013-05-30|2017-05-02|Burcon Nutrascience Corp.|Production of pulse protein products with reduced astringency|WO2017127927A1|2016-01-26|2017-08-03|Burcon NutrascienceCorp.|Ph adjusted pulse protein product|

---

US9635875B2|2013-05-30|2017-05-02|Burcon NutrascienceCorp.|Production of pulse protein products with reduced astringency|

---

US20160159868A1|2014-10-01|2016-06-09|Burcon NutrascienceCorp.|Production of soy protein products with reduced astringency |

---

WO2016049763A1|2014-10-01|2016-04-07|Burcon NutrascienceCorp.|Production of soy protein products with reduced astringency |

---

CN107404897B|2015-03-18|2021-10-22|不二制油集团控股株式会社|Method for producing chocolate food and improvement thereof|

---

CA2980561A1|2015-03-24|2016-09-29|Cargill, Incorporated|Corn protein isolate and methods of manufacturing same|

---

CN109153982A|2016-01-07|2019-01-04|睿普食品公司|The component and its preparation process of product analog or this kind of analog|

---

CA3012532A1|2016-01-27|2017-08-03|Burcon NutrascienceCorp.|Preparation of non-soy oilseed protein products |

---

CA3054856A1|2017-03-03|2018-09-07|Burcon NutrascienceCorp.|Preparation of acid soluble pulse protein hydrolyzates with little or no astringency and pulse protein hydrolyzates of improved amino acid score|

---

US11102998B1|2017-08-25|2021-08-31|The Hershey Company|Binders and methods of making and using the same|

---

US10143226B1|2018-01-15|2018-12-04|Innovative Proteins Holding, LLC|Yellow pea protein compositions with high digestibilities and amino acid scores|

---

EP3540035A1|2018-03-13|2019-09-18|The Procter & Gamble Company|Hand dishwashing detergent composition|

---

CA3108620A1|2018-09-17|2020-03-26|Societe Des Produits Nestle S.A.|Non-dairy drink with rice and pea proteins|

---

AU2018447545A1|2018-11-04|2021-04-01|Societe Des Produits Nestle S.A.|Thermally treated composition comprising plant proteins and methods of production and use thereof|

---

AU2020294519A1|2019-06-18|2022-02-03|Corn Products Development, Inc.|Pulse protein emulsifiers|

---

US20220071260A1|2020-09-08|2022-03-10|Ripple Foods, Pbc|Solid shaped food containing non-dairy protein|

法律状态:
2018-02-27| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

---

2018-03-27| B15K| Others concerning applications: alteration of classification|Ipc: A23J 1/14 (2006.01), A23J 3/14 (2006.01), A23J 3/1 |

---

2019-09-10| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

---

2020-07-21| B07A| Application suspended after technical examination (opinion) [chapter 7.1 patent gazette]|

---

2020-12-08| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

---

2021-02-09| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 30/05/2014, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

---

申请号 | 申请日 | 专利标题

---

US201361828735P| true| 2013-05-30|2013-05-30|

---

US61/828,735|2013-05-30|

---

US201461927182P| true| 2014-01-14|2014-01-14|

---

US61/927,182|2014-01-14|

---

PCT/CA2014/000466|WO2014190418A1|2013-05-30|2014-05-30|Production of pulse protein products with reduced astringency|

---