CN112126668B - Method for preparing bitter-free soybean polypeptide by specific endo-complex enzyme - Google Patents
Method for preparing bitter-free soybean polypeptide by specific endo-complex enzyme Download PDFInfo
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- 230000007062 hydrolysis Effects 0.000 claims description 66
- 238000006460 hydrolysis reaction Methods 0.000 claims description 66
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- 238000004925 denaturation Methods 0.000 claims description 14
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- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims description 13
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- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
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- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
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- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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Abstract
The invention discloses a method for preparing bitter-free soybean polypeptide by specific endo-complex enzyme, which comprises the following steps: s1, adding water into soybean protein isolate, and uniformly stirring and mixing; s2, regulating the PH value of the mixture obtained in the step S1, and heating to denature the protein; s3, regulating the pH value of the mixture obtained in the step S2 to 4.0-9.0, and adding specific endo-compound protease to hydrolyze isolated soy protein to obtain an enzymolysis liquid at the temperature of 40-60 ℃; compared with the preparation method on the market, the preparation method of the soybean peptide powder has the advantages of low total enzyme addition amount, simple operation, less required process equipment and low production cost.
Description
Technical Field
The invention belongs to the technical field of vegetable protein processing, and mainly relates to a method for preparing bitter-free soybean polypeptide by using specific endo-compound enzyme.
Background
The soybean protein is internationally recognized ideal plant protein, the polypeptide overcomes the weakness of the soybean protein in nutrition, and the soybean protein has more abundant nutrition and functional characteristics than the soybean protein. The soybean polypeptide has become an effective auxiliary preparation, can be used as a nutrient source of microorganisms in the fermentation industry, promotes the growth and metabolism of the microorganisms, and can be applied to baked foods, candy cakes and cold drink foods in the food industry, thereby reducing the cost and improving the flavor, taste, tissue state and the like of the product; meanwhile, in recent years, soybean polypeptide is easy to absorb and digest, can reduce cholesterol, inhibit blood pressure rise, has low antigenicity, can enhance immunity and resist free radical loss and other physiological functions, and has important significance in developing novel functional health-care foods of peptides.
In the prior art, two main problems encountered in the preparation of soybean polypeptide are that when in vitro enzymatic digestion of soybean protein is carried out, the soybean protein has compact structure and contains a large amount of hydrophobic amino acids, deep hydrolysis is difficult to realize, and the enzymatic conversion rate is greatly lower than that of milk-derived protein and fish protein, so that the production cost is high; meanwhile, due to hydrolysis, hydrophobic amino acid residues in the soybean protein molecule are exposed, and the types and arrangement sequences of the amino acid residues of the bitter peptide are caused, the obtained soybean polypeptide product has serious bitter taste, and the application of the soybean polypeptide is limited. In order to remove the bitter taste of the soybean peptide product, various debittering processes including adsorption, masking, etc. have been developed, and additional debittering processes inevitably complicate the process, increase the cost, and lower the yield.
The existing method for preparing soybean polypeptide mainly comprises a microbial fermentation method and an enzymolysis method, wherein the enzymolysis method comprises a single-enzyme hydrolysis method, a double-enzyme hydrolysis method, a compound enzyme enzymolysis method and the like. Along with the deep research, people gradually recognize the defects of the single-enzyme hydrolysis method, the multi-enzyme composite hydrolysis method gradually becomes an application key point, and the performance of the prepared product is generally higher than that of the single-enzyme hydrolysis method.
Disclosure of Invention
The invention aims to solve the defects in the prior art, and provides a method for preparing the soybean polypeptide without bitter taste by using the specific endo-compound enzyme, so as to achieve the purposes of reducing the production cost, improving the hydrolysis degree of soybean protein, removing the bitter taste of the soybean polypeptide and realizing the efficient utilization of the byproduct of the aqueous enzymatic method.
The technical scheme of the invention is as follows: a method for preparing a bitter-free soybean polypeptide by using a specific endo-complex enzyme, comprising the following steps:
s1, adding water into soybean protein isolate, and uniformly stirring and mixing;
s2, regulating the PH value of the mixture obtained in the step S1, and heating to denature the protein;
s3, regulating the pH value of the mixture obtained in the step S2 to 4.0-9.0, and adding specific endo-compound protease to hydrolyze isolated soy protein to obtain an enzymolysis liquid at the temperature of 40-60 ℃;
s4, heating the enzymolysis liquid for deactivation treatment, wherein the heating temperature is 80-100 ℃ and the heating time is 10-30 minutes;
s5, filtering and separating the enzymolysis liquid after the inactivation treatment to obtain clear hydrolysis filtrate, and recovering unhydrolyzed soy protein isolate for secondary hydrolysis;
s6, concentrating and desalting the clarified hydrolysis filtrate obtained in the step S5 by utilizing nanofiltration membrane equipment; desalting, evaporating and concentrating, and pulverizing to obtain non-bitter soybean polypeptide powder.
The ratio of the mass of the soy protein isolate in S1 to the volume of water is 8% -15%.
In the step S2, the pH value of the feed liquid is regulated to 3.0-6.0, the protein denaturation temperature is controlled to be 70-95 ℃, and the denaturation time is controlled to be 10-20 minutes.
The specific endo-compound protease in the S3 is the compound enzymolysis of proline endoprotease and/or aspartic endoprotease and/or serine endoprotease and acid protease and/or alkaline protease and/or neutral protease and/or beer enzyme;
the addition amount of the proline endoprotease, the aspartic endoprotease, the serine endoprotease, the acid protease, the alkaline protease, the neutral protease and the beer enzyme is 0.2-9% of the ammonia endoprotease and/or 0.2-9% of the aspartic endoprotease and/or 0.2-9% of the serine endoprotease and 0.2-9% of the acid protease and/or 0.2-9% of the alkaline protease and/or 0.2-9% of the neutral protease and/or 0.2-9% of the beer enzyme, the total addition amount of the specific endoprotease accounts for 0.2-9% of the mass percentage of the isolated soy protein, and the hydrolysis time is 2-7 hours.
The pH values of the feed liquid in the S2 and the S3 are regulated by sulfuric acid solution with mass fraction of 30-50% or sodium hydroxide solution with mass fraction of 30-50%.
And (2) the hydrolysis degree of the clarified hydrolysis filtrate in the step (S5) is 30-85%, unhydrolyzed soybean protein isolate is recovered, the collected filter residues are subjected to water re-dissolution cleaning, filtration separation is repeated for 1-4 times, the collected treated filter residues are substituted into the soybean protein isolate powder with the mass fraction of 75% -150%, and the secondary recovery hydrolysis degree is 20% -64%.
And (3) desalting and concentrating in the step (S6) to obtain yellow or dark yellow clear filtrate, and freeze-drying or spray-drying to obtain powder, thereby obtaining the soybean peptide powder product with the molecular weight mainly distributed in 500D-2000D.
The beneficial effects of the invention are as follows: the preparation method of the invention relates to the steps of raw material pretreatment, enzymolysis reaction, reaction termination, product purification, secondary recovery of reaction residues and the like. The raw material treatment is that the soybean protein isolate raw material is treated by adding acid at high temperature, so that the space structure of soybean protein is destroyed, and the enzyme treatment time is shortened; then carrying out compound hydrolysis by adopting 1-7 proteases with different characteristics; stopping the enzyme reaction by adopting high-temperature treatment after reaching a proper hydrolysis degree; filtering and separating the reacted feed liquid, desalting, concentrating and pulverizing to obtain soybean peptide powder product; and (5) filtering the filter residues, cleaning, reducing swelling reaction, and recycling for secondary use.
The preparation method of the soybean peptide powder has lower total enzyme addition compared with the preparation method on the market. And the pH value is not required to be adjusted by sectional enzymolysis during hydrolysis, the pH value and the temperature are only required to be adjusted once after the raw material pretreatment, and the one-pot method feeding can start the hydrolysis to the enzymolysis to finish the inactivation treatment, so that the operation is simple, the required process equipment is less, and the production cost is low.
On the basis of the process for preparing the soybean polypeptide by the complex enzymolysis of the aqueous enzymatic method, the invention adopts proline endoprotease, aspartic endoprotease, serine endoprotease complex acid protease, neutral protease, alkaline protease, beer enzyme and the like, and the specific enzymolysis of the soybean protein isolate ensures that the hydrolysis degree is high; the hydrolysis rate is high; the cut polypeptide has low hydrophobicity and the proportion of hydrophobic amino acid residues such as Lys, val, leu, pro, phe, tyr, ile, trp and the like in the polypeptide is low so as to remove the bitter taste of the soybean peptide powder. The technology of pretreatment denaturation of soybean protein isolate, direct feeding by a one-pot method, concentration and desalination of enzymolysis clear liquid and the like are adopted, so that the physicochemical property and structural characteristic of soybean protein are improved, and the quality and content of soybean polypeptide are improved. The method has the characteristics of simple process equipment, safe operation, secondary hydrolysis utilization of residue recovery, less total enzyme addition, low production cost and high production benefit, and the prepared soybean polypeptide has good quality, no beany flavor and no bitter taste.
Detailed Description
The present invention will be further illustrated with reference to specific examples, but the present invention is not limited to the examples. The means used in the examples, unless otherwise specified, are all means conventional in the art.
Example 1:
dissolving soybean protein isolate in pure water to obtain 8% (W/V) water-powder mixture, adjusting pH to 3 (regulated by 50% sulfuric acid), heating to 80deg.C, and denaturing for 10 min; then adjusting the pH value of the mixed solution after the protein denaturation treatment to 9 (adjusting by 50% sodium hydroxide), wherein the temperature is 40 ℃, adding proline endoprotease (self-production), aspartic endoprotease, serine endoprotease, acidic protease, alcalase alkaline protease (novelin) and neutral protease which account for 6% of the mass percentage of the soybean protein isolate powder, wherein the total complex enzyme addition accounts for 6% of the mass percentage of the soybean protein isolate powder, and the hydrolysis time is 5 hours. And heating the enzymolysis liquid for deactivation treatment after hydrolysis, wherein the heating temperature is 85 ℃ and the heating time is 30 minutes. Cooling the inactivated material to below 35 ℃, filtering to separate clear hydrolysate with the hydrolysis degree of 33%, recovering unhydrolyzed soybean protein isolate, adding water to the collected filter residue, re-dissolving, cleaning, filtering and separating for 2 times, mixing the collected filter residue with soybean protein isolate powder with the mass fraction of 100%, and continuously recycling, wherein the secondary recovery hydrolysis degree is 22%. Desalting the clarified hydrolysate with nanofiltration membrane, evaporating and concentrating the desalted solution to obtain solid 20, and freeze drying to obtain soybean peptide powder.
Example 2:
dissolving soybean protein isolate in pure water to obtain water-powder mixture with 9% (W/V) ratio, adjusting pH to 4 (regulated by 40% sulfuric acid), heating to 90deg.C, and denaturing for 15 min; then adjusting the pH value of the mixed solution after the protein denaturation treatment to 8 (adjusting by 40% sodium hydroxide), wherein the temperature is 40 ℃, and adding proline endoprotease (self-production), aspartic endoprotease, serine endoprotease, acid protease, alcalase alkaline protease (novelin) and neutral protease which account for 0% of the mass percentage of the soybean protein isolate powder, wherein the total compound enzyme addition accounts for 6% of the mass percentage of the soybean protein isolate powder, and the hydrolysis time is 5 hours. And heating the enzymolysis liquid for deactivation treatment after hydrolysis, wherein the heating temperature is 85 ℃ and the heating time is 30 minutes. Cooling the inactivated material to below 35 ℃, filtering to separate clear hydrolysate with the hydrolysis degree of 35%, recovering unhydrolyzed soybean protein isolate, adding water to the collected filter residue, re-dissolving, cleaning, filtering and separating for 2 times, mixing the collected filter residue with soybean protein isolate powder with the mass fraction of 90%, and continuously recycling, wherein the secondary recovery hydrolysis degree is 23%. Desalting the clarified hydrolysate with nanofiltration membrane, evaporating and concentrating the desalted solution to obtain solid 20, and spray drying to obtain soybean peptide powder.
Example 3:
dissolving soybean protein isolate in pure water to obtain 8% (W/V) water-powder mixture, adjusting pH to 4 (regulated by 40% sulfuric acid), heating to 80deg.C, and denaturing for 10 min; then adjusting the pH value of the mixed solution after the protein denaturation treatment to be 5 (adjusting by 50% sodium hydroxide), wherein the temperature is 45 ℃, adding proline endoprotease (self-production) accounting for 0% of the mass percentage of the soybean protein isolate powder, aspartic endoprotease 0% serine endoprotease 0%, acid protease 6%, alcalase alkaline protease 0% (Novalosin), neutral protease 0% and beer enzyme 0%, wherein the total complex enzyme addition accounts for 6% of the mass percentage of the soybean protein isolate powder, and the hydrolysis time is 5 hours. And heating the enzymolysis liquid for deactivation treatment after hydrolysis, wherein the heating temperature is 85 ℃ and the heating time is 30 minutes. Cooling the inactivated material to below 35 ℃, filtering to separate clear hydrolysate with the hydrolysis degree of 43%, recovering unhydrolyzed soybean protein isolate, adding water to the collected filter residue, re-dissolving, cleaning, filtering and separating for 2 times, mixing the collected filter residue with soybean protein isolate powder with the mass fraction of 110%, and continuously recycling, wherein the secondary recovery hydrolysis degree is 32%. Desalting the clarified hydrolysate with nanofiltration membrane, evaporating and concentrating the desalted solution to obtain solid 20, and freeze drying to obtain soybean peptide powder.
Example 4:
dissolving soybean protein isolate in pure water to obtain 8% (W/V) water-powder mixture, adjusting pH to 4 (adjusted by 45% sulfuric acid), heating to 80deg.C, and denaturing for 10 min; then adjusting the pH value of the mixed solution after the protein denaturation treatment to 9 (adjusting by 50% sodium hydroxide), wherein the temperature is 43 ℃, adding proline endoprotease (self-production) accounting for 0% of the mass percentage of the soybean protein isolate powder, aspartic endoprotease 0% serine endoprotease 0%, acid protease 0%, alcalase alkaline protease 6% (Novalosin), neutral protease 0% and beer enzyme 0%, wherein the total complex enzyme addition accounts for 6% of the mass percentage of the soybean protein isolate powder, and the hydrolysis time is 5 hours. And heating the enzymolysis liquid for deactivation treatment after hydrolysis, wherein the heating temperature is 85 ℃ and the heating time is 30 minutes. Cooling the inactivated material to below 35 ℃, filtering to separate clear hydrolysate with the hydrolysis degree of 46%, recovering unhydrolyzed soybean protein isolate, adding water to the collected filter residue, re-dissolving, cleaning, filtering and separating for 2 times, mixing the collected filter residue with soybean protein isolate powder with the mass fraction of 100%, and continuously recycling, wherein the secondary recovery hydrolysis degree is 34%. Desalting the clarified hydrolysate with nanofiltration membrane, evaporating and concentrating the desalted solution to obtain solid 20, and spray drying to obtain soybean peptide powder.
Example 5:
dissolving soybean protein isolate in pure water to obtain 10% (W/V) water-powder mixture, adjusting pH to 5 (regulated by 30% sulfuric acid), heating to 95deg.C, and denaturing for 15 min; then adjusting the pH value of the mixed solution after the protein denaturation treatment to 9 (adjusting by 30% sodium hydroxide), wherein the temperature is 45 ℃, adding proline endoprotease (self-production) accounting for 4% of the mass percentage of the soybean protein isolate powder, aspartic endoprotease 0%, serine endoprotease 0%, acid protease 0%, alcalase alkaline protease 4%, neutral protease 0% and beer enzyme 0%, wherein the total complex enzyme addition accounts for 8% of the mass percentage of the soybean protein isolate powder, and the hydrolysis time is 5 hours. And heating the enzymolysis liquid for deactivation treatment after hydrolysis, wherein the heating temperature is 85 ℃ and the heating time is 30 minutes. Cooling the inactivated material to below 35 ℃, filtering to separate clear hydrolysate with the hydrolysis degree of 56%, recovering unhydrolyzed soybean protein isolate, adding water to the collected filter residue, re-dissolving, cleaning, filtering and separating for 2 times, mixing the collected filter residue with soybean protein isolate powder with the mass fraction of 110%, and continuously recycling, wherein the secondary recovery hydrolysis degree is 40%. Desalting the clarified hydrolysate with nanofiltration membrane, evaporating and concentrating the desalted solution to obtain solid 20, and spray drying to obtain soybean peptide powder.
Example 6:
dissolving soybean protein isolate in pure water to obtain water-powder mixture with 11% (W/V) ratio, adjusting pH to 4 (regulated by 50% sulfuric acid), heating to 90deg.C, and denaturing for 15 min; then adjusting the pH value of the mixed solution after the protein denaturation treatment to 8 (adjusting by 40% sodium hydroxide), wherein the temperature is 50 ℃, adding proline endoprotease (self-production) accounting for 2% of the mass percentage of the soybean protein isolate powder, aspartic endoprotease 2%, serine endoprotease 0%, acid protease 0%, alcalase alkaline protease 2%, neutral protease 0% and beer enzyme 0%, wherein the total complex enzyme addition accounts for 6% of the mass percentage of the soybean protein isolate powder, and the hydrolysis time is 6 hours. And heating the enzymolysis liquid for deactivation treatment after hydrolysis, wherein the heating temperature is 85 ℃ and the heating time is 30 minutes. Cooling the inactivated material to below 35 ℃, filtering to separate clear hydrolysate with the hydrolysis degree of 60%, recovering unhydrolyzed soybean protein isolate, adding water to the collected filter residue, re-dissolving, cleaning, filtering and separating for 2 times, mixing the collected filter residue with soybean protein isolate powder with the mass fraction of 120%, and continuously recycling, wherein the secondary recovery hydrolysis degree is 45%. Desalting the clarified hydrolysate with nanofiltration membrane, evaporating and concentrating the desalted solution to obtain solid 20, and freeze drying to obtain soybean peptide powder.
Example 7:
dissolving soybean protein isolate in pure water to obtain water-powder mixture with 12% (W/V) ratio, adjusting pH to 5 (regulated by 40% sulfuric acid), heating to 85deg.C, and denaturing for 15 min; and then adjusting the pH value of the mixed solution after the protein denaturation treatment to 8 (adjusting by 50% sodium hydroxide), wherein the temperature is 55 ℃, adding proline endoprotease (self-production) accounting for 2% of the mass percentage of the soybean protein isolate powder, 0% of aspartic endoprotease, 2% of serine endoprotease, 1% of acid protease, 2% of Alcalase alkaline protease (novelin), 0% of neutral protease and 0% of beer enzyme, wherein the total complex enzyme addition accounts for 7% of the mass percentage of the soybean protein isolate powder, and the hydrolysis time is 5 hours. And heating the enzymolysis liquid for deactivation treatment after hydrolysis, wherein the heating temperature is 85 ℃ and the heating time is 30 minutes. Cooling the inactivated material to below 35 ℃, filtering to obtain clear hydrolysate with the hydrolysis degree of 63%, recovering unhydrolyzed soybean protein isolate, adding water to the collected filter residue, re-dissolving, cleaning, filtering, separating for 4 times, mixing the collected filter residue with soybean protein isolate powder with the mass fraction of 120%, and continuously recycling, wherein the secondary recovery hydrolysis degree is 46%. Desalting the clarified hydrolysate with nanofiltration membrane, evaporating and concentrating the desalted solution to obtain solid 20, and spray drying to obtain soybean peptide powder.
Example 8:
dissolving soybean protein isolate in pure water to obtain water-powder mixture with 13% (W/V) ratio, adjusting pH to 5 (regulated by 50% sulfuric acid), heating to 80deg.C, and denaturing for 20 min; then adjusting the pH value of the mixed solution after the protein denaturation treatment to 9 (adjusting by 40% sodium hydroxide), wherein the temperature is 50 ℃, adding proline endoprotease (self-production) accounting for 1% of the mass percentage of the soybean protein isolate powder, aspartic endoprotease 1%, serine endoprotease 1%, acid protease 1%, alcalase alkaline protease 1%, neutral protease 1% and beer enzyme 1%, wherein the total complex enzyme addition amount accounts for 7% of the mass percentage of the soybean protein isolate powder, and the hydrolysis time is 4 hours. And heating the enzymolysis liquid for deactivation treatment after hydrolysis, wherein the heating temperature is 85 ℃ and the heating time is 30 minutes. Cooling the inactivated material to below 35 ℃, filtering to separate clear hydrolysate with the hydrolysis degree of 74%, recovering unhydrolyzed soybean protein isolate, adding water to the collected filter residue, re-dissolving, cleaning, filtering and separating for 2 times, mixing the collected filter residue with soybean protein isolate powder with the mass fraction of 130%, and continuously recycling, wherein the secondary recovery hydrolysis degree is 52%. Desalting the clarified hydrolysate with nanofiltration membrane, evaporating and concentrating the desalted solution to obtain solid 20, and freeze drying to obtain soybean peptide powder.
Example 9:
dissolving soybean protein isolate in pure water to obtain water-powder mixture with a ratio of 14% (W/V), adjusting pH to 5 (adjusted by 50% sulfuric acid), heating to 85deg.C, and denaturing for 15 min; then adjusting the pH value of the mixed solution after the protein denaturation treatment to 9 (adjusting by 50% sodium hydroxide), wherein the temperature is 55 ℃, adding proline endoprotease (self-produced) accounting for 1% of the mass percentage of the soybean protein isolate powder, aspartic endoprotease 1.5%, serine endoprotease 1%, acid protease 1%, alcalase alkaline protease (novelin) 1.5%, neutral protease 1.5% and beer enzyme 0.5%, wherein the total complex enzyme addition accounts for 8% of the mass percentage of the soybean protein isolate powder, and the hydrolysis time is 5 hours. And heating the enzymolysis liquid for deactivation treatment after hydrolysis, wherein the heating temperature is 85 ℃ and the heating time is 30 minutes. Cooling the inactivated material to below 35 ℃, filtering to separate clear hydrolysate with the hydrolysis degree of 81%, recovering unhydrolyzed soybean protein isolate, adding water to the collected filter residue, re-dissolving, cleaning, filtering and separating for 3 times, mixing the collected filter residue with soybean protein isolate powder with the mass fraction of 130%, and continuously recycling, wherein the secondary recovery hydrolysis degree is 58%. Desalting the clarified hydrolysate with nanofiltration membrane, evaporating and concentrating the desalted solution to obtain solid 20, and spray drying to obtain soybean peptide powder.
The following are a part of the experimental results:
table 1 comparison of physicochemical index of examples
It can be seen from the comprehensive table 1 that the soybean peptide powder prepared by the specific endo-compound enzyme has the advantages of high hydrolysis degree, low bitter taste, high recycling rate and the like compared with the soybean peptide powder prepared by the single-enzyme hydrolysis method and the double-enzyme hydrolysis method.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (4)
1. A method for preparing a bitter-free soybean polypeptide by using a specific endo-complex enzyme, which is characterized by comprising the following steps:
s1, adding water into soybean protein isolate, stirring and uniformly mixing to prepare a water-powder mixture with the proportion of 14% (W/V);
s2, regulating the PH value of the mixture obtained in the step S1, and heating to denature the protein;
s3, regulating the pH value of the mixture obtained in the step S2 to 9.0, adding specific endo-compound protease to hydrolyze the isolated soy protein to obtain enzymolysis liquid, adding proline endoprotease accounting for 1 percent of the mass of the isolated soy protein powder, aspartic endoprotease accounting for 1.5 percent of the mass of the isolated soy protein powder, serine endoprotease accounting for 1 percent of the mass of the isolated soy protein powder, acid protease accounting for 1 percent of the mass of the isolated soy protein powder, alkaline protease accounting for 1.5 percent of the mass of the isolated soy protein powder, neutral protease accounting for 1.5 percent of the mass of the isolated soy protein powder and beer enzyme accounting for 0.5 percent of the mass of the isolated soy protein powder, adding total compound enzyme to the mass of the isolated soy protein powder for 8 percent of the mass of the isolated soy protein powder for 5 hours,
s4, heating the enzymolysis liquid for deactivation treatment, wherein the heating temperature is 80-100 ℃ and the heating time is 10-30 minutes;
s5, filtering and separating the inactivated enzymolysis liquid to obtain clear hydrolysis filtrate, wherein the hydrolysis degree of the clear hydrolysis filtrate is 30-85%, unhydrolyzed soybean protein isolate is recovered, water re-dissolving and cleaning are carried out on the collected filter residues, filtering and separating are repeated for 1-4 times, and the soybean protein isolate powder with the mass fraction of 75-150% is mixed with the collected filter residues and is substituted into S1 for recycling, and the secondary recovery hydrolysis degree is 20-64%;
s6, concentrating and desalting the clarified hydrolysis filtrate obtained in the step S5 by utilizing nanofiltration membrane equipment; desalting, evaporating and concentrating, and pulverizing to obtain non-bitter soybean polypeptide powder.
2. The method for preparing soybean polypeptide without bitter taste by using specific endo-complex enzyme according to claim 1, wherein the pH value of the feed liquid is adjusted to 3.0-6.0 in S2, the protein denaturation temperature is controlled at 70-95 ℃, and the denaturation time is controlled at 10-20 minutes.
3. The method for preparing the bitter-free soybean polypeptide by using the specific endo-complex enzyme according to claim 1, wherein the pH value of the feed liquid in S2 and S3 is regulated by using a sulfuric acid solution with the mass fraction of 30-50% or a sodium hydroxide solution with the mass fraction of 30-50%.
4. The method for preparing soybean polypeptide without bitter taste by specific endo-complex enzyme according to claim 1, wherein the yellow or deep yellow clear filtrate is obtained after desalting and concentrating in the step S6, and the soybean peptide powder product with molecular weight mainly distributed in 500D-2000D is obtained after freeze drying or spray drying into powder.
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