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    • 专利摘要:
    O F I N V E N T I O N The present invention relates to an immobilized saccharifying enzyme product in particulate form, said saccharifying enzyme being selected from the group consisting of amyloglucosidase and maltogenic alpha-amylase, which product comprises carrier particles consisting mainly of casein granules essentially coated with a liquid-permeable proteinaceous layer in which the saccharifying enzyme is co-crosslinked with egg albumen by means of glutaraldehyde, and the invention also relates to a process for the preparation of an immobilized saccharifying enzyme product in particulate form, said saccharifying enzyme being selected from the group consisting of amyloglucosidase and maltogenic alpha-amylase, which process comprises essential= ly coating carrier particles consisting mainly of precipitated casein with a proteinaceous layer in which the saccharifying enzyme is co-crosslinked with egg albumen by means of glutaral= dehyde.
    公开号:SU921470A3
    申请号:SU772503856
    申请日:1977-07-01
    公开日:1982-04-15
    发明作者:Амотц Шмуль；Кер Нильсен Таге；Берге Росениус Поульсен Поуль；Эдмунд Норман Барри
    申请人:Ново Индустри А/С (Фирма)；
    IPC主号:
    专利说明:

    (54) THE SYMAGES OF OBTAINING AN IMMOBILIZED OSACHARIZING ENZYME DRUG. : The invention relates to methods for the preparation of insoluble enzyme preparations,. in particular, preparations derived from saccharifying enzymes - amnloglucose and a-a dylase, which are used in the food industry and adjacent fields. The process of transformation into insoluble cottage (or immobilization) of extracellular and intracellular water-soluble enzymes, i.e., fixation of catalytically active (Naive) enzymes in solid or quaevite, molded or possessing the ability to form structures, becomes more and more important technologically as well as economically, it can be considered as a means, allowing it to reuse the ground enzyme in batch processes or use it in uninterrupted so called enzyme reactors. In recent years, considerable efforts have been directed towards the development of immobilization technology in the starch industry, which is associated with the production of products such as highly concentrated dextrose and fructose syrups. Recently, in the food and related industries, the relative importance of the second of these two products has increased, with highly concentrated fructose syrups mostly replacing sucrose syrup and less sweet highly concentrated dextrose syrup. Two additional types of sugar syrup obtained by hydrolysis of starch and containing a significant amount of maltose (i.e. syrups with a high content of maltose and a high degree of conversion) are used to an ever increasing extent, especially in confectionery (hard candy) and the canning industry. The complete industrial process of transforming starch through a high dextrose high-fat skillet into a high fructose syrup includes three successive stages, namely the process of diluting or liquefying starch. Acid-catalyzed or bacterial amylase dextrins, followed by 392 saccharifying them to high dextrose syrup and conversion of the latter to high fructose syrup, each process being catalyzed by its specific enzyme, namely amyl lucosidase and glucose isomerase, respectively. While, as a result of a series of recent technological developments, it became possible to carry out the first and third stages of this sequence continuously, industrial saccharification (t, e, second stage) is still a batch process. high-maltose content can also be obtained from starch.The saccharification of liquefied starch, which results in a syrup with a high content of maltose (and usually containing ositelno slight amount of glucose) is performed via malyogennoy amylase, preferably a fungal a-amylase. Like the glycogen saccharification of the corresponding maltogen), the 1st process is usually carried out as a batch process using a soluble enzyme. Although the preference for a completely uninterrupted process is obvious, the improvement of the designs of an industrial enzymatic reactor, based on the use at the saccharification stage of immobilized amylocosidase, is at the initial stage. Several methods are known for immobilizing saccharifying enzymes, for example, amyloglucosidase, by binding the enzyme to an insoluble organic or inorganic carrier. However, only a few of these methods went beyond laboratory practice. For example, a method is known for the preparation of an immobilized saccharifying enzyme preparation based on amyloglucosidase by covalently linking it with porous particles of glass or ceramic materials using glutaric aldehyde. Particles of a carrier with an average pore diameter of about 400 A are placed in a column and subsequently a solution of glutaraldehyde and an enzyme solution is passed through it. The resulting product is used in a continuous saccharification process. The column of the pilot plant was filled with the obtained ground enzyme enzyme and poured with commercially available dextrins. A degree of conversion to dextrose is achieved, which is close to 92%, which is usually realized in the processes of periodic saccharification with soluble amyloglucosidase. However, in contrast to the batch process, in which the free enzi case is used, in the column the degree of vizimorezorotvany, i. The increased interconversion encountered when using an immobilized enzyme preparation of this type is apparently due to the use of porous enzyme carriers. Obviously, the porous structure greatly increases the total surface area, causing the maximum binding of enzymes, and therefore the maximum enzymatic activity of the immobilized enzyme preparation obtained (1. However, the combination of a high concentration of enzyme bound to the surface of the carrier and a lower diffusion rate pore limits inevitably leads to locally high glucose concentrations, which creates favorable conditions for accelerating enzyme processes with high K values (i.e., undesirable interconversion reactions, especially those that lead to the formation of isomaltose and isomaltriose). The aim of the invention is to improve the quality of the immobilized enzyme preparation obtained, causing a reduction in the formation of by-product polymerization when using the obtained preparation in the saccharification process. At the same time, the performance characteristics of the resulting product should be substantially close to those of the soluble enzyme, regardless of whether Whether the immobilized drug is in a batch process or in a continuous scheme process. In the case of an immobilized amyloglucosidase preparation, this requirement leads to the fact that a continuous saccharification process can be carried out under such particularly preferred conditions (for example, with such composition and dextrin feed rate) at which the glucose concentration at the output is 92%, with the total disaccharide concentration ( maltose and isomaltose) and trisaccharides (mainly panoses and isomaltotriose) would not exceed approximately 4 and 1%, respectively. In the case of immobilized maltogenic a-amylase, the necessary conversion parameters are as follows: maltose, about 40-60%; maltotriosis 25-35% and glucose less than 10%. This goal is achieved by the fact that, according to the method of obtaining immobilized
    the saccharifying enzyme preparation of binding the enzyme to an insoluble carrier using glutaraldehyde granular casein is used as a carrier, the binding process is carried out with s stirring in an aqueous medium or in the presence of egg albumin in the following ratios: enzyme: carrier (0.02 - 0,2) enzyme: albumin (0,2-1,5) glutaraldehyde: enzyme with albumin (02, -0,4): and the water content in the reaction mixture is 30-60 wt.%, Followed by mechanical, crushing the resulting product and drying.
    As a saccharifying enzyme, amyloglucosidase or o-amylase is used.
    Glutaraldehyde is usually used in the form of a 50% aqueous solution.
    The preferred particle size of casein is 00-500 microns.
    The product obtained according to the invention is a carrier particle, i.e., granular casein, coated with a protein layer of albumin, permeable to liquid, and in this layer, the saccharifying enzyme is cross-linked with egg albumin with glutaric aldehyde.
    The method is carried out as follows.
    A thoroughly mixed dry mixture of 30 granulated casein and egg albumin powder is combined with an aqueous mixture containing saccharifying enzyme and glutaraldehyde, dissolved at pH 4-7, and albumin can be arbitrarily dissolved 35 with these components instead of mixing it in a dry state with casein The resulting mixture is quieted in a quiescent state, usually at ambient temperature, in order to complete the coating of the carrier with the 40 protein layer and the crosslinking process of the protein. The displacement can be carried out either manually, for example, by thorough mixing and grinding in a mortar, by thorough mixing and grinding the protrusion, or mechanically, for example, in a horizontal dump mixer of the type supplied by the form of the People’s Machinebau, Zaadadna Germany, or similar type of industrial mixers. 50, The water content in the mixture should be 30-60 wt.%.
    The properties of the granular casein product used as a carrier material are important. So, granules. casein must have considerable physical strength in order to be able to withstand the significant deformations that occur when soaking the nozzle column.
    In addition, the degree of swelling in water should be reasonably 1ZHZKO, preferably should not exceed 200%. An example of a product that satisfies these requirements is granulated acid precipitated casein. Food grades of products of this type, preferably having a particle size of 00-500 microns, can be obtained from various sources, for example, they are produced by the French company Szerm C. Either scanning the surface of such particles is very curved and unstable lano minimizing macroscopic pumice. It is likely that such a povervechnoe microstructure has a large number of binding 5 asthkas for glutaraldehyde.
    For the implementation of the invention, it is important that albumin used as a binding protein dissolves almost instantly in water. Hence, the preferred albumin variety is the usual spray dried product.
    during the mixing process, a certain degree of agglomeration of the coated particles may occur, which leads to the completion of the cross-linking reaction during the formation of a wet, coarse-grained and lumpy product. This product must be further crushed, for example, by granulating to form a crushed product with a particle size in the preferred range. Granulation can be carried out on an oscillating one: a granulator of the type that is supplied by a number of Companies (Diaf, Copenhagen, or Manosti, Liverpool). The granular product is passed through a sieve, for example, with a hole size of 1-2 mm, and it can be freed from the thin material by standard methods. The resulting product can be npoMbiT and then dried to the desired water content. I.
    Before using in the process of saccharification, the dried enzyme product is usually Uluoyasht, for which it is pre-soaked in an aqueous solution. At this stage, as well as at the initial stage of subsequent use, a certain decrease in enzyme activity can occur. Found that the inclusion of an additional stage of cross-linking. arbitrarily combined with the soaking process can lead to a significant decrease in the indicated loss of enzyme activity. Hence, in accordance with a preferred embodiment, the invention pretreats the enzyme preparation with an aqueous solution containing an acceptable amount of glutaraldehyde, preferably 0.5 to 5%, followed, as a rule, by removing excess reagent by pulling and removing the dried preparation. In addition, it was found that by pretreatment of the enzymatic product with a salt of sulfurous acid, a significant increase in the degree of conversion (expressed as percentage of dextrose equivalent or the value of EH) of the saccharified product can be achieved. The sulfite effect is very likely due to the disclosure of certain mesh structures, including the protein enzymatic active layer, but, apparently, without binding the enzyme, which thereby facilitates the access of higher molecular weight oligosachrids to the active sites of the enzyme. Thus, the treatment is preferable (enzymatic product with a dilute aqueous solution of sulfite containing 0.1-2% sodium sulfite at pH 4-5. The sulfite treatment is preferably carried out at room temperature and is usually interrupted after 120 minutes by washing with water, followed by extraction of the dried product. It has been established that, after the treatment with sulfite, there is no loss of enzyme activity. Enzyme preparations prepared according to the described procedure can be used in the process of saccharification or period action, including the separation and reuse of an enzyme preparation, or a continuous action in an enzymatic reactor. Amyloglucosidase is obtained by cultivating the membranes belonging to the species Aspergillus or Rizopus, i.e. Aspergillus niger or Rizopus delemar. oryzea. Determination of amyloglucosidase activity Under standard conditions, i.e. at pH 4.5 and temperature 55.0 ° C, the specific activity of soluble amyloglucosidase is determined, as well as the specific activity of immobil The enzyme is an enzyme, which is defined as the amount of an enzyme or enzyme product that hydrolyzes an aqueous solution containing 30% (wt.) of maltose, with an initial rate of 1 micromolar maltose per minute. The activity of the immobilized enzyme is determined by the process of periodic action on the sample, which is maintained in a suspended state by means of an agitator. Determination of the activity of fungal a-amyl Under standard conditions, namely at pH 5.6-5.7, temperature 37, oio, 05 ° C and in the presence of 4.3 mmol of Ca ions, the specific activity of the soluble a- amylase, as well as the specific activity of the immobilized enzyme, expressed as the amount of enzyme or fermentative product that hydrolyzes the solution containing soluble starch (Merck brand Ami mind solubile, DAV, Erg. 8. Vf), (6.95 g per 000 ml) starting rate of 5.26 mg to ahmala per hour. To control the reaction, blue color with iodine is observed. When the starch emulsion breaks down, this color becomes weaker and gradually turns into reddish-brown. The change in color is controlled visually by comparison with the color 1 glass standards. The activity of the immobilized enzyme is tested in terms of conditions analogous to those indicated by the data of myoglucosidase. In soluble saccharifying enzymes, industrial Novo ISO amyglucosidase 1600 and fungamyl 1600 are used (fungicidal patents; trade name of the product). The first, which is sold in the form of an aqueous solution, is usually converted to a spray dried powder before use. The drying process may be preceded by ultrafiltration, usually combined with a washing procedure to remove low molecular weight impurities. By applying the processes of concentration and purification, well known to those skilled in the art, a solid amyloglucosidase product with an activity of 5000 units can be easily obtained. per gram or higher activity. Fungamil 1600, marketed at. powder form, has an activity of 1600 units. per gram It can be used directly as a commercial drug, and the drug subjected to additional concentration and purification. The complete extraction of activity in an immobilized saccharifying enzyme npojoyKTe depends on the specific parameters of the immobilization process, but is usually not less than 40%. The substrate in the process of saccharification, carried out using an immobilized enzyme preparation in accordance with the invention, can be any oligosaccharide that undergoes saccharification in the presence of the corresponding soluble enzyme. Examples of preferred substrates are dextrins obtained by acidic and / or enzymatic dilution of starch and characterized by DE-values equal to 5-40; highly concentrated maltose syrups (with EH values from 35 to 60) to the residual oligosaccharides remaining in the mother liquor after dextrose utah crystallization in raffinates obtained by fractionating fructose and glucose. Preferred dextrin solutions with a dry matter content ranging from 20 to 55% (wt.ab.). - c-m and m with p 1. Relatively low-grade granulated casein precipitated with hydrochloric acid (9 g pshtsevogo casein, sold by the firm Szerma SA, France, and containing particles of 100-500 µm in size, and the fraction from 300 to 500 m is about 60%) is mixed with industrial albumin albumin, dried (1.2 g). To this mixture is added a pre-mixed solution of amyloglucosidase powder (6.5 tons of an 18.5% aqueous solution) containing a total of 12,500 units. amyloglucosidase and glutaraldehyde (1.2 ml of a 50% aqueous solution). Amyloglucosidase powder is an ultrafiltered, flow-dried product prepared from Novy amyloglucosidase (Novo Industry A / S, Dani). The mixture is thoroughly ground in a mortar and then kept at room temperature for 1 hour. The resulting aggregates are ground in a mortar to obtain a granulated product, which is allowed to dry at room temperature for one day. The activity of the dried granulated product (11.4 g) is 425 units. per gram, which is 38.8%. Example 2. Acid-granulated precipitated casein (1500 g of type M 60 Cs. Serum A.A. containing particles of 100-500 µm in size, the fraction from 150 to 350 µm being about 70%) are mixed in a dry state with commercially dried jet albumin (120 g) in a 20-liter horizontal mixer. The ultrafiltered amyloglucosidase concentrate prepared from Novo amyloglucosidase (650 g of a solution containing about 25% dry matter and about 3200 units of soluble amyloglucosidase in 1 G of concentrate) is thoroughly mixed with glutaric aldehyde (180 ml of a 50% aqueous solution) at pH 4 , 9 and at a temperature, the resulting solution is poured into the mixer. Thorough mixing is carried out for another half minute or one minute, and then the wet precipitated product is removed from the mixer. The wet product is left at rest for about 45 minutes to complete the crosslinking process, resulting in chunks of aggregated particles. The granulation of the product is carried out on an oscillating granulator (for example, of the type supplied by Diaf A / S, Kopeetagen), equipped with screens with 1.5 mm apertures. The granulated product is washed with desalted water (10 l) for 10 minutes, removed by filtration and then subjected to drying in a fluidized bed. The dried product (1600 g, 450 units of amyloglucosidase per gram) is freed from fine particles by sieving. Example 3. A solution of 105 g of a spray-dried powder of amyloglucosidase prepared in Example 1 and 105 g of spray-dried albumin was prepared in 400 ml of tap water and left to stand overnight in a refrigerator at 6-7 ° C, pH of 5.4 . Glutaraldehyde (100 ml of a 50% aqueous solution) is then added and the resulting solution is added to the well-stirred for 3 minutes (a portion of the granulated hydrochloric acid precipitated casein (725 g of the same sort as used in Example 1) The mixture was mixed in an industrial mixer of the type used in Example 2. The mixture was thoroughly stirred for several minutes, after which it was left to rest for about 1 hour to complete the crosslinking reaction. The resulting aggregates and the lump product are granulated on an oscillating granulator through a 2 mm sieve. The granulated product is washed with a solution of sodium acetate (2pH 4.2) and then dried under vacuum at 35 ° C. The activity of the dried granulated product (925 g) is 800 units. per gram, which corresponds to 67.8% of the initial activity. Example 4. The immobilized amyloglucosidase product prepared according to Example 3 (20 g) was soaked at room temperature for 1 hour in an aqueous solution of glutaraldehyde (500 ml of a 1% solution adjusted to pH 7.0), after which extraction of the dried product (20 g).::,. . The activity of the obtained product is 625 unit grams, that is, in the second stage of immobilization, the relative activity is 7%, and the total recovery from soluble amyloglucosidase is 53%. , Example 5, An aqueous solution of maltogenic alpha-amylase (2 ml of a 5% solution of unyamyl 1600 (Novo Industry A / S, Dani) with deionized water at pH 6.2) is mixed with an aqueous solution of glutaric aldeide (0.3 ml of 50% solution) and then added quickly to a dry mixture of acid-granulated precipitated casein (4/0 g type M 60) and industrial egg albumin, spray-dried (0.5 g). 119 After thorough grinding in a mortar, the obtained product is kept at 1 at room temperature. The pieces are crushed in a mortar after which the product sugiat at room temperature for one day. Example 6. A column with a diameter of 15 m equipped with a bruise and maintained at 55 ° C was charged with 5.7 g of an immobilized amyloglucosidase product prepared according to prnmer 1. The column was fed at a constant rate of 15 ml per hour from top to bottom, containing 30% (weight vol.) acidified with (an enzyme of hydrolyzed dextrin (CPC iCHoy Flzik Maltodextrin 01915 DE), having the following approximate value,%: Glucose4-5 Disaccharides. 8-9 Trisaccharides6-7 Tetra and oligosaccharides 79-82 k to which 0.2% (w / w%) of sodium sulfite is added and then the pH is adjusted to 4.5. The flow was analyzed by high pressure liquid chromatography (HPLC). The data obtained are presented in Table 1. Example 7. A 25 mm diameter immobilized amyloglucosidase product (20 g) was loaded into a column with a diameter of 25 mm and maintained at 55 ° C. prepared according to example 2. Raw materials of the same concentration, the same composition and pH, which is used in example 6, are fed from the bottom up to the column. The feed rate is constant and equal to 50 ml. at one o'clock. The glucose content is determined by the becoxinase method. The data obtained are listed in Table 2. Example 8. A column with an internal diameter of 25 mm, equipped with a jacket and under f f. immobilized amyloglucosidase product (25 g), prepared according to example 2, the composition feedstock is fed from bottom to top. used in example 6. Raw materials (pH 4.5) contains dry solids 30% (wt. wt.) and with the addition of 2 g per liter of potassium sorbate as an antioxidant. The flow rate is adjusted in this way to maintain an approximately constant degree of conversion. The effluent is analyzed by the HPLC. The amount of glucose conversion is determined over time. The data presented in Table. 3. After the experiment was completed, no changes in the hardness and other physical properties of the material of the packing of the column were noted. Example 9. Following the procedure of Example 7, but replacing immobilization with amyloi12 glucose / sludge product of Example 2 with that prepared in Example 4, the results are presented as shown in the following table. 4. Example 10. Immobilized maltogenic alpha-amylase prepared according to the method of example 5 (4 g) is loaded onto a srubber column maintained at a temperature of 4–5 ° C and the raw material — industrial dextrin (ATP Snow Flake Maltodextrkn 0191.3) is fed from the top down, containing Wf (wt .ab.) solid solids and 0, sodium sulfite as an antioxidant. The pH of the raw material is adjusted to 4.5 and the flow rate is set at 15 ml per hour. 5 indicates the composition of the stream flowing from the column, with the disaccharides represented mainly by maltose. PRI me R 11. A). A column with an internal diameter of 15 mm, equipped with a shirt, which is maintained at 55 ° C, is loaded with 12 g of immobilized amyloglucosidase prepared according to the procedure of Example 2. I The dextrin feedstock is fed from top to bottom (31% (w / w), calculated on the dry matter content) prepared by diluting starch with Termamyl L 60 (trade name, Novo Industry A / S, Dani), and having the following composition,%: Glucose1 Shs & Harishl. 7-8 Trisccharides .10-12 Tetra and oligosaccharides79-82 D. Е.21 0.2% (w / v) sodium sulfite is added to the feedstock as an antioxidant, and the feedstock has a pH of 4. The flow is adjusted to thus, to maintain a PERMANENT degree of conversion (Consumer from prodrug glucose). The effluent is analyzed by the HPLC. In tab. 6 presents the results obtained. at). The dextrin substrate prepared according to the method described in section (A) and having the following composition,%: Glucose1.1 Disaccharides6.8 Trisadrides11.0 Tetra and oligosaccharides: 81.0 are treated with 0.01% (based on the content of dry of the hard substance) maltogenic alpha-amylase (Fungamil 1600, Novo-Industry A / S) at pH 5.0 and for 2 hours. After the treatment, the following composition of the substrate is crawled,%: Glucose1.3 Disaccharides13.9 Trisaccharides22.3 Tetra- and oligosaccharides62,5 Each of two columns with an internal diameter of 15 mm, with a shirt and a sub at a temperature of 55 ° C, immobilized amyloglucosidase (10 g prepared according to Example 2) is loaded .. In columns n, the raw material is supplied from top to bottom, and the raw material contains 30% (w / v) of two types of dextrin prepared according to this description; 0.2% (potassium sorbate w / v and pH is set to 4.5. 1u {the maximum DH value at the output of the processing of dextrin with fungamil is 92.7%, and the maximum value of the DH value at the output after processing dextrin with fungal 93.3%. Example 12. A 10 g portion of the immobilized amylogyucosidase prepared by the procedure of Example 2 is soaked in 100 m solutions containing various concentrations of sodium sulfite. The pH of the solution is adjusted to 4.5 by adding acetic acid. After soaking for 2 hours at room temperature, the enzyme samples are washed with deionized water and loaded into columns with an inner diameter of 15 mm, jacketed and maintained at 55 ° C. The columns are fed from top to bottom with streams of raw material containing dextrin of 30% (w / v) prepared according to the procedure of Example PA and to which 0.2% (w / v) of potassium sorbate is added, followed by pH adjustment to 4.5 . By adjusting the flow rate in each column, the following results are obtained from the condition of maximizing the DH value at the outlet: Sodium sulfite. Maximum DH g / 100 ml% O90.3 0.0 f90.2, 0.191.2 0.291.7 0.591.9 1.092 , 4 Example 13. The stream collected at the outlet of the column with immobilized fungamil of example 10 with a high content of maltose (DE approximately 42) is used as the raw material for the column of example b with immobilized amyloglucose daza. The flow rate is adjusted (up to 73 ml per hour) so that the output will have a high conversion syrup (DE approximately 65), which has the following condition,%: Glucose 38.1 Disaccharides39.6 Trisaccharides2.8 Tetra and oligosaccharides19, 6 Example 14. A immobilized amyloglucosidase product (20 g) prepared according to Example 3 is loaded into a column having an internal diameter of 25 mm and equipped with a crushing, at a temperature of 55 ° C. Raw materials containing mother liquor (or raffinate) are fed upwards from the bottom of the column, sex) Fructose-glucose fractionation P1enu, having the following tav,%: Fructose: 3 25 Glucose 85.71 Disaccharides9.17 Trisaharvda, 42 Higher saccharides0.54 Moreover, the pH is adjusted to 4.5, and the flow rate through the column is 250 ml per hour at a concentration of 25% (weight.weig .). The composition at the outlet of the column, determined by the HPLC, is as follows: Fructose3.25 Glucose91.16 Disaccharides4.14 Trisaccharides1.15 Higher saccharides0.30 The advantage of the present invention is the relatively low cost of the preparations obtained, since their preparation is based on relatively low-cost commercially easily produced osteel and other auxiliary materials.
    5.6 4.2
    92.0 93.2
    3.9
    92 3.2 93.6
    3.1
    933 3.2 92.6
    2.9
    92.6
    I
    Days
    one
    .
    1.7 2.0
    95.3 95.8
    2.5
    95.4 95.8
    2.5
    2.9 3.3
    95.5 94.9
    94
    3.6
    Table 2 The glucose content,%
    89.0 93.6 93.6 93.1
    Table. 3
    Glucose content.
    101
    92.4 Flow rate, ml / h
    17
    9 10
    Days
    13
    921470
    18 Continuation of table 3
    60 57
    92.0
    Table 4
    Glucose content
    93.3
    3.6
    92.3
    3.6
    92.2 92.4 3.1
    3.3
    92.1
    56
    权利要求:
    Claims (4)
    [1]
    Claim I. Investigation of an immobilized saccharifying enzyme preparation by linking a saccharifying enzyme saccharide with an insoluble carrier using glutaraldehyde, characterized in that, in order to improve the quality of the target product. granulated casein is used as a carrier, and binding is carried out with stirring in an aqueous medium with the additional presence of egg albumin in the following ratios of components: enzyme: carrier (0.02-0.2) enzyme: albumin (0.2-1.5) glutarium- aldehyde: enzyme with albumin (0.2-0.4): and containing 30–60 wt.% of water in the reaction mixture, followed by mechanical crushing of the obtained product and drying.
    3.3
    33
    3.5
    27 18 3.6
    3.7
    15
    [2]
    2. The method according to claim 1, dtl and h ay rs and and -; so that amyloglucosidase is used as a saccharifying enzyme or
    “-Amylase.
    [3]
    3. The method according to claim 1, wherein the particle size of the carrier is 00-500 microns.
    [4]
    4. The method according to any one of the preceding claims, so that glutaraldehyde is isolar in the form of a 50% aqueous solution.
    Sources of information taken into account in the examination
    -, I .-;
    1. D. D, Lee, J. J. Lee, G. T. Tsao. Continuons Production of Glucose from Dektrin by Glucoamylase tmmobitized on Porous SJIica DieStarke-, 1975. Nr. 1, 27. 384-387 (about .totype).
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    同族专利:
    公开号 | 公开日
    ES460304A1|1978-10-01|
    DK146417C|1984-03-05|
    ZA773580B|1978-05-30|
    AR210710A1|1977-08-31|
    IT1112120B|1986-01-13|
    DK292177A|1978-01-03|
    AU508098B2|1980-03-06|
    JPS6117470B2|1986-05-07|
    FR2356665B1|1983-07-22|
    GB1571987A|1980-07-23|
    AU2619677A|1978-12-21|
    CA1072029A|1980-02-19|
    DK146417B|1983-10-03|
    NL7707233A|1978-01-04|
    MX4281E|1982-03-11|
    JPS539389A|1978-01-27|
    FR2356665A1|1978-01-27|
    BR7704346A|1978-04-11|
    DE2729459A1|1978-01-12|
    BE856365A|1978-01-02|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    RU2711790C1|2018-12-26|2020-01-22|федеральное государственное бюджетное образовательное учреждение высшего образования "Воронежский государственный университет" |Method of producing polybromoline preparation using glutaric aldehyde|NL6812443A|1968-08-31|1970-03-03|
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    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    GB27749/76A|GB1571987A|1976-07-02|1976-07-02|Enzyme products|
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