• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Procedure for the sequential collection of high added value compounds from wet biomass. The present invention relates to a process that allows the recovery of the different compounds of high added value (proteins, lipids, pigments and carbohydrates) from wet biomass by successive extractions with subcritical pressurized water, with expanded ethanol with CO 2 sub- or supercritical and, finally, with CO 2 supercritical. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2695874A1
    申请号:ES201730896
    申请日:2017-07-05
    公开日:2019-01-11
    发明作者:EZEQUIEL Mª ELENA IBÁÑEZ；López Bienvenida Gilbert；León José Antonio Mendiola；Gallego Alejandro Cifuentes；Calleja Miguel Herrero
    申请人:Consejo Superior de Investigaciones Cientificas CSIC；
    IPC主号:
    专利说明:

    [0001] Procedure for the sequential collection of high added value compounds from wet biomass
    [0002]
    [0003]
    [0004] The present invention relates to a process that allows the recovery of the different compounds of high added value (soluble proteins, lipids, pigments and carbohydrates) from wet biomass through successive extractions with subcritical pressurized water, with ethanol expanded with sub-CO2. or supercritical and, finally, with supercritical CO2.
    [0005]
    [0006] STATE OF THE ART
    [0007] One of the most important challenges in the development of bio-refineries of microalgae is the improvement of processes that allow the integral recovery of the compounds of high added value from the algal biomass (proteins, carbohydrates, lipids, pigments, phenolic compounds) . Although much progress has been made in this field, practically all processes start with the drying of the biomass after harvesting (to eliminate the water content, up to 80%) using drying procedures, such as lyophilization , and that require high energy and operating costs.
    [0008]
    [0009] For example, the document: Welcome Gilbert-Lopez, Jose A. Mendiola, Javier Fontecha, Lambertus AM van den Broek, Lolke Sijtsma, Alejandro Cifuentes, Miguel Herrero and Elena Ibáñez, Green Chem., 2015, 17, 4599, presents a process in four sequential stages for the extraction of different biologically functional compounds such as soluble proteins, lipids and pigments from microalgae. Prior to the extraction process in 4 stages, we proceed with a lyophilization procedure of the microalgae.
    [0010]
    [0011] The high energy cost of these drying processes of the algal wet biomass is reflected in the following document: Hou Jian, Yang Jing, and Zhang Peidong, The Scientific World Journal, Volume 2015 (2015), Article ID 920968, 9 pages, where the difference in energy expenditure between extracting lipids from dry algae or wet algae is explained by subcritical co-solvents, achieving an improvement of 43.83% in the energy ratio of wet algae on dry algae. Therefore, less energy is consumed to extract compounds from wet algae than from dry algae.
    [0012] Additionally the patent US9328310 (B1) cites a process for the extraction of lipids from wet algae with subcritical water. For this, in a preferred embodiment, the document indicates that a cake of wet biomass, which is not freeze-dried, and which is constituted by algae, is pressurized in a reactor and proceeds with an extraction with liquid under pressure, using water as solvent .
    [0013]
    [0014] As a consequence of the documents studied, these processes of extraction of biomass formed by algae or microalgae, present several disadvantages such as: the high energy expenditure associated with the drying processes in some cases, as well as the expense associated with the number of operations or extractions necessary to achieve high percentages of obtaining high added value compounds that are extracted from the starting biomass in others.
    [0015]
    [0016] Therefore, it is necessary to develop an extraction procedure to take advantage of all the high added value and biologically functional compounds present in the biomass (for example, algal biomass) and to reduce the energy expenditure associated with the different processes involved in the process. same and the pre-treatments necessary to perform the extractions as they have been done so far.
    [0017]
    [0018] DESCRIPTION OF THE INVENTION
    [0019]
    [0020] Surprisingly, sequentially performing extractions with compressed fluids from directly wet biomass, such as algae, which is developed in the present invention, not only eliminates the need for drying or lyophilization of the starting biomass that is carried out in the procedures of the state of the art, with the pertinent energy saving, but also leads to a percentage yields of the extracted compounds higher than expected or those described in the state of the art following other procedures than the one of the present invention.
    [0021]
    [0022] Therefore, a first aspect of the present invention relates to a process for the recovery of high value added compounds from wet biomass comprising the following steps:
    [0023]
    [0024] (a) extraction with subcritical water at a pressure between 1-11 MPa and a temperature between 30-50 ° C to obtain a first extract with high added value compounds and a residue, and subsequent drying of the obtained residue;
    [0025]
    [0026] (b) extraction of the residue obtained in step a) by an extraction process at a pressure of 5-10 MPa and a temperature of between 40-60 ° C with sub- or supercritical CO2 (ScCO2) using a percentage of ethanol between 40-80%, to obtain a second extract with high added value compounds and a second residue, and then drying the second obtained residue; Y
    [0027]
    [0028] (c) finally, extraction of the second residue obtained in step b) by means of a supercritical CO2 extraction process at a pressure between 25-60 MPa and at a temperature between 40-70 ° C to obtain a third extract with compounds of high added value and a third residue.
    [0029]
    [0030] In a preferred embodiment the starting moist biomass can be frozen or at a temperature comprised between 10-30 ° C.
    [0031]
    [0032] In another preferred embodiment the compounds of high added value are proteins, lipids (polar, medium polarity and low polarity) and pigments (carotenoids, xanthophylls and chlorophylls) and carbohydrates. Preferably the proteins extracted are those soluble in water and ethanol.
    [0033]
    [0034] In another preferred embodiment of the process, the extracts obtained in steps a), b) and c) are collected in vessels protected from light.
    [0035]
    [0036] In another preferred embodiment of the described process, the wet biomass comprises algal biomass, microalgae or mixture of the above.
    [0037]
    [0038] In another preferred embodiment of the process, the ethanol of the extract obtained in step b) is removed under reduced pressure and temperature between 25-35 ° C, thus preventing the ethanol from degrading the high added value compounds extracted in said step b ).
    [0039] In another preferred embodiment of the process, the extract obtained in step a) is frozen to avoid degradation of the high added value compounds extracted in said step a).
    [0040]
    [0041] In another preferred embodiment, the moist biomass comprises microalgae, and more preferably the microalgae are selected from the list comprising Isochrysis galbana, Scenedesmus obliquus, Phaeodactylum tricornutum, Nannochloropsis gaditana, Anacystis nidulans, Neochloris oleoabundans Spirulina, Dunaliella, Haematococcus and any mixture thereof. previous
    [0042]
    [0043] In another preferred embodiment of the described process, the drying of the residues of steps a) and b) is carried out in N2 at a pressure of less than 1.5 MPa in a time of between 10 and 70 minutes. More preferably the drying of the residues of steps a) and b) is carried out in N2 at a pressure of between 0.03 and 1 MPa for a time of between 20 and 60 minutes.
    [0044]
    [0045] In another preferred embodiment of the described process, the subcritical water extraction of step a) is carried out at a pressure of between 9-11 MPa and a temperature of between 30-40 ° C. More preferably, the subcritical water extraction of step a) is carried out at a pressure of 10 MPa and a temperature of 30 ° C.
    [0046]
    [0047] In another preferred embodiment of the described process, the extraction using a percentage of expanded ethanol of between 40-80% with sub- or supercritical CO2 of stage b) is carried out at a pressure of between 6.5-8 MPa and a temperature of between 45 and 55 ° C. More preferably, extraction using a percentage of expanded ethanol of between 40-80% with CO2 from step b) is carried out at a pressure of 7 MPa and a temperature of 50 ° C.
    [0048]
    [0049] In another preferred embodiment of the described process, the extraction with supercritical fluids of step c) is carried out at a pressure between 25-60 MPa and at a temperature between 60-70 ° C. More preferably, the extraction with supercritical fluids of step c) is carried out at a pressure of 30 MPa and a temperature of 70 ° C.
    [0050]
    [0051] In a preferred embodiment, a step prior to step (a) of the process described in the present invention is carried out, characterized in that the biomass wet is mixed with a filling under temperature conditions between 10 ° C and 30 ° C and in a weight ratio between wet biomass and filling between 1: 5 and 1:20, where the filling is an inert material selected from the list comprising glass balls, Raschig rings, Fenske rings, sea sand, silica gel of 60 Ánstróng pore size and silica gel of pore size of 120 Ánstróng. In this case it will be the wet biomass next to the filling that is introduced in stage (a) to perform the extraction with supercritical water.
    [0052]
    [0053] In the present invention, Raschig rings are understood to be pieces of tubular geometry whose diameter is approximately equal to their length and which are used as filling for columns in distillation processes and in other engineering chemical processes.
    [0054]
    [0055] In the present invention, Fenske rings refer to pieces of glass in the form of a helix with a defined diameter and which are used as fillers for columns in distillation processes and in other engineering chemical processes.
    [0056]
    [0057] In the present invention, as is well known to a person skilled in the art, silica gel means a granular and porous form of silicon dioxide synthetically manufactured from sodium silicate. Despite the name, silica gel is solid.
    [0058]
    [0059] In the present invention, "wet biomass" is understood to mean one containing a content percentage by weight of water of more than 20% by weight, greater than 40%, greater than 60% and more preferably a water content by weight of between 65% and 90%.
    [0060]
    [0061] In the present invention "algal biomass" is understood as the total amount of material obtained from algae and / or microalgae cultures in open reactors, ponds, photobioreactors or any type of process used for their production and growth. invention the algal biomass has been supplied by Fitoplancton Marino SL (Cádiz, Spain), and that are used as raw material of the extraction processes described in the present invention.
    [0062]
    [0063] In the present invention, the obtained extracts are preferably preserved from light to avoid degradation of the high added value compounds obtained.
    [0064] In the present invention, "extraction with compressed fluids" is understood to mean those in which the extractant used is a compound subjected to high pressure, and comprises any of the following techniques well known as: Extraction with supercritical fluids (SFE), extraction with liquids pressurized (PLE), such as with pure water or pure ethanol, and extraction under subcritical conditions or with expanded liquids.
    [0065]
    [0066] Throughout the description and the claims the word "comprises" and its variants do not intend to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and characteristics of the invention will emerge partly from the description and partly from the practice of the invention. The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention.
    [0067]
    [0068] The Project that leads to this patent application has received its funding from the Seventh Framework Program for research, technological development and demonstration under the grant agreement no. 613588.
    [0069]
    [0070] BRIEF DESCRIPTION OF THE FIGURES
    [0071]
    [0072] FIG. 1 Histogram representing the recovery percentage of proteins, carbohydrates, lipids and pigments for the procedure in three stages for each of these stages.
    [0073]
    [0074] FIG. 2 Histogram representing the recovery percentage of proteins, carbohydrates, lipids and pigments for the procedure in four stages of the bibliography for each of these stages.
    [0075]
    [0076] EXAMPLES
    [0077]
    [0078] The invention will now be illustrated by means of tests carried out by the inventors, which highlights the effectiveness of the product of the invention.
    [0079]
    [0080] Example 1: Process of sequential extraction of high added value compounds from wet algal biomass according to the method of the present invention
    [0081] The procedure of sequential extraction of a sample of 25 g of wet biomass (with a humidity of 72%) comprising I. galbana in a supercritical fluid extractor "Spe-ed Helix" is carried out. 300 ml basket and it is placed in the high pressure stainless steel extraction cell, pressurized CO2 is pumped for the pre-conditioning of the chamber and the pressurized biomass cake is preheated to the extraction temperature.
    [0082]
    [0083] The extraction is carried out in three sequential steps using as solvents of each extraction (i) pure water (PLE) (ii) an extraction of expanded liquid, of ethanol expanded with supercritical CO2 and (iii) an extraction with supercritical CO2 (SFE), with a drying with N2 at low pressure between each of the extractions.
    [0084]
    [0085] The extraction with pressurized liquid using water as extractant (i) is carried out by feeding the cell with water until reaching a pressure of 10 MPa, and maintaining a temperature of 30 ° C. The extraction of polar compounds contained in the sample of the microalgae is carried out in a single static cycle for 30 minutes. Subsequently, the biomass sample is dried with N2 at 1 MPa for 40 minutes.
    [0086]
    [0087] Extraction of expanded liquid, in particular of ethanol expanded with CO2 (ii), is carried out by pumping ScCO2 up to 7 MPa at 50 ° C, at the same time that the extraction cell is fed with a flow rate of 2.5 ml / min of ethanol. The extraction is performed dynamically for a period of 90 minutes. Subsequently, the biomass sample is dried with N2 at 1 MPa for 30 minutes.
    [0088]
    [0089] Extraction with supercritical fluids with ScCO2 (iii) is carried out by pumping ScCO2 up to 30 MPa at 70 ° C in the extraction cell. The extraction is performed dynamically for a period of 90 minutes.
    [0090]
    [0091] All the collection containers of each of the extractions are protected from light. Finally, for the best preservation of the extracted compounds, on the one hand, for the extract of the second stage with ethanol expanded with ScCO 2 , the ethanol is eliminated by means of reduced pressure and temperature of 30 ° C, and, on the other hand, the extract obtained in the first stage of extraction in water under pressure, must be frozen.
    [0092] Example 2: Sequential extraction procedure of high added value compounds from dry algal biomass (lyophilized)
    [0093]
    [0094] By way of comparison, a dry algal biomass extraction process is related in four successive stages using compressed fluids:
    [0095]
    [0096] The procedure of sequential extraction of a sample of 10 g of lyophilized biomass comprising I. galbana in a supercritical fluid extractor "Spe-ed Helix" is carried out.The sample is mixed homogeneously with 30 g of sea sand and the whole is placed in a 300 ml basket that is placed in the high pressure stainless steel extraction cell, pressurized CO2 is pumped for the pre-conditioning of the chamber and the pressurized biomass cake is preheated to the extraction temperature .
    [0097]
    [0098] The extraction is carried out in four sequential steps using as solvents of each extraction (i) supercritical CO2 (ii) an extraction of expanded liquid, of ethanol expanded with supercritical CO2 (iii) an extraction with pure pressurized ethanol (PLE) and (iv) an extraction with pure water.
    [0099]
    [0100] Extraction with supercritical fluid with ScCO2 (i) is carried out by pumping ScCO2 up to 30 MPa at 50 ° C in the extraction cell. The extraction is performed dynamically for a period of 60 minutes.
    [0101]
    [0102] The extraction with an expanded liquid, in particular of ethanol expanded with CO2 (ii), is carried out by pumping ScCO2 up to 7 MPa at 50 ° C at the same time that the extraction cell is fed with a flow rate of 2.5 ml / min of ethanol. The extraction is performed dynamically for a time of 120 minutes.
    [0103]
    [0104] The extraction with pressurized liquid using ethanol as extractant (iii) is carried out by feeding the cell with water until reaching a pressure of 10 MPa, and maintaining a temperature of 80 ° C. The extraction of compounds of medium-high polarity contained in the sample of microalgae is carried out in a single static cycle for 30 minutes.
    [0105] Subsequently, the biomass sample is dried with N2 at 1 MPa for 30 minutes.
    [0106]
    [0107] The extraction with pressurized liquid using water as extractant (iv) is carried out by feeding the cell with water until reaching a pressure of 10 MPa, and maintaining a temperature of 80 ° C. The extraction of polar compounds contained in the sample of the microalgae is carried out in a single static cycle for 30 minutes.
    [0108]
    [0109] All the collection containers of each of the extractions are protected from light. Finally, for the best preservation of the extracted compounds, on the one hand, the extracts of the second and third stages (with expanded ethanol with ScCO2 and pure ethanol, respectively), the ethanol is eliminated by rotavapor (at reduced pressure), and by On the other hand, the extract obtained in the fourth stage of extraction in pressurized water must be frozen.
    [0110]
    [0111] Example 3: Comparison of the yields of high added value compounds obtained in the processes of examples 1 and 2 described above
    [0112]
    [0113] Recovery obtained for each type of compound in the extract, with respect to the initial amount in the alga, obtained for each stage of the extraction process for the 4-stage and 3-stage procedure.
    [0114]
    [0115] Table 1: Shows the yields (in% by weight with respect to the percentage of each compound in the initial algal biomass sample) of each of the high value-added compounds obtained in the procedure of Example 1 (with wet biomass) and 2 (with dry biomass).
    [0116]
    [0117]
    [0118]
    [0119]
    [0120] n represents the number of repetitions of the entire process.
    [0121] SD is defined as the relative standard deviation
    [0122] As can be seen in table 1, the yields obtained in the process corresponding to the present invention are clearly superior to those obtained by the process from dry (lyophilized) biomass known in the state of the art. For quantified compounds of interest (lipids, proteins, sugars and the fucoxanthin pigment), recoveries are higher, reaching values higher than 80% of recovery of the lipids present in the algal biomass and 70% of fucoxanthin, both extracted mainly in the stages (ii) and (iii) of the sequential process. Regarding the content of proteins and sugars, the process corresponding to the present invention allows to recover a percentage of proteins higher than 18% and a percentage of sugars greater than 35%, mainly in stages (i) and (ii) of the sequential process .
    [0123]
    [0124] The residue of the sequential extraction process contains mainly sugars and proteins and corresponds to the percentage not extracted in the sequential stages.
    [0125]
    [0126] Additionally, in the absence of a previous drying or lyophilization stage in the process of the present invention, this implies an improvement in the level of energy consumption with respect to the process known in the state of the art.
    权利要求:
    Claims (17)
    [1]
    1. Procedure for obtaining sequence! of high value added compounds from wet biomass comprising the following stages:
    (a) extraction of the wet biomass with subcritical water at a pressure of between 1-11 MPa and a temperature of between 30-50 ° C to obtain a first extract that comprises high added value compounds and a residue, and subsequent drying of that residue obtained;
    (b) extraction of the dry residue obtained in step (a) by an extraction process at a pressure of 5-10 MPa and a temperature of between 40-60 ° C with sub- or supercritical CO2 using an ethanol percentage between 40 -80%, to obtain a second extract comprising high added value compounds and a second residue, and then drying that residue; Y
    (c) extraction of the dry residue obtained in step (b) by means of a supercritical CO2 extraction process at a pressure between 25-60 MPa and at a temperature between 40-70 ° C to obtain a third extract comprising compounds of high added value and a third residue.
    [2]
    Process according to the preceding claim, wherein the high value added compounds of each of the extracts obtained in steps (a), (b) and (c) are selected from proteins, lipids, pigments, carbohydrates or combinations thereof.
    [3]
    3. Method according to any of the preceding claims wherein the extracts obtained in steps (a), (b) and (c) are collected in vessels protected from light.
    [4]
    4. Process according to any of the preceding claims wherein the ethanol of the extract obtained in step b) is removed under reduced pressure and temperature between 25-35 ° C.
    [5]
    Process according to any of the preceding claims wherein the extract obtained in step (a) is frozen.
    [6]
    6. Process according to any of the preceding claims wherein the wet biomass comprises algal biomass, microalgae or any mixture of the above.
    [7]
    Method according to any of the preceding claims wherein the wet biomass comprises microalgae selected from the list comprising Isochrysis galbana, Scenedesmus obliquus, Phaeodactylum tricornutum, Nannochloropsis gaditana, Anacystis nidulans, Neochloris oleoabundans, Spirulina, Dunaliella, Haematococcus and any mixture of the above .
    [8]
    8. Process according to any of the preceding claims wherein the wet biomass is frozen or at a temperature comprised between 10 and 30 ° C.
    [9]
    9. Process according to any of the preceding claims wherein the drying process of steps (a) and (b) is carried out with N2 at a pressure of less than 1.5 MPa in a time of between 10 and 70 minutes.
    [10]
    The process according to claim 9, wherein the drying process of steps (a) and (b) is carried out in N2 at a pressure of between 0.03 and 1 MPa for a time of between 20 and 60 minutes.
    [11]
    The process according to any of the preceding claims wherein the subcritical water extraction of step (a) is carried out at a pressure of between 9 11 MPa and a temperature of between 30-40 ° C.
    [12]
    The process according to claim 11, wherein the subcritical water extraction of step (a) is carried out at a pressure of 10 MPa and a temperature of 30 ° C.
    [13]
    Method according to any of the preceding claims, wherein the extraction using a percentage of expanded ethanol of between 40-80% with CO2 from step (b) is carried out at a pressure of between 6.5-8 MPa and a temperature of between 45-55 ° C.
    [14]
    The process according to claim 13, wherein the extraction using a percentage of expanded ethanol of between 40-80% with CO2 from stage (b) is carried out at a pressure of 7 MPa and a temperature of 50 ° C.
    [15]
    15. Method according to any of the preceding claims wherein the supercritical CO2 extraction of step (c) is carried out at a pressure between 25-60 MPa and at a temperature between 60-70 ° C.
    [16]
    16. The process according to claim 15, wherein the supercritical CO2 extraction of step (c) is carried out at a pressure of 30 MPa and a temperature of 70 ° C.
    [17]
    Method according to any one of the preceding claims, wherein a step prior to stage (a) is carried out, characterized in that the wet biomass is mixed with a filling under temperature conditions between 10 ° C and 30 ° C and in a weight ratio between the wet biomass and the filling between 1: 5 and 1:20, where the filling is an inert material selected from the list consisting of glass balls, Raschig rings, Fenske rings, sea sand and gel of silica of pore size between 60 and 120 Ánstróng.
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    同族专利:
    公开号 | 公开日
    WO2019008211A1|2019-01-10|
    ES2695874B2|2019-06-07|
    引用文献:
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    WO2014052823A1|2012-09-27|2014-04-03|Yale University|System and method for separating lipid based products from biomass|
    CN111214846A|2019-11-29|2020-06-02|云南绿A生物产业园有限公司|Haematococcus pluvialis extract and preparation method thereof|
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    ES201730896A|ES2695874B2|2017-07-05|2017-07-05|Procedure for the sequential collection of high added value compounds from wet biomass|ES201730896A| ES2695874B2|2017-07-05|2017-07-05|Procedure for the sequential collection of high added value compounds from wet biomass|
    PCT/ES2018/070482| WO2019008211A1|2017-07-05|2018-07-05|Method for the sequential production of high-added-value compounds from wet biomass|
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