CN104073540A - Polypeptide production method for enhancing utilization ratio of protein - Google Patents
Polypeptide production method for enhancing utilization ratio of protein Download PDFInfo
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Abstract
The invention relates to a polypeptide production method which sequentially comprises the following steps: (1) taking a protein-containing raw material, adding water, and immersing to obtain a first solution; (2) stirring and homogenizing the first solution; (3) carrying out enzymolysis on the first solution with alkaline proteinase; (4) inactivating and centrifugating the enzymolysis solution obtained in the step (3) to obtain a supernate A and a precipitate A; (5) adding water to the precipitate A to obtain a second solution; carrying out enzymolysis on the second solution with neutral proteinase; (6) inactivating and centrifugating the enzymolysis solution obtained in the step (5) to obtain a supernate B and a precipitate B; (7) adding water to the precipitate B to obtain a third solution; carrying out enzymolysis on the third solution with acidic protease; (8) inactivating the enzymolysis solution obtained in the step (7), and centrifugating to obtain a supernate C; (9) mixing the supernate A, supernate B and supernate C to obtain a polypeptide mixed solution; and (10) removing water in the polypeptide mixed solution to obtain the polypeptide. The method can enhance the protein utilization ratio of the raw material and obtain the high-quality polypeptide product.
Description
Technical field
The present invention relates to the preparation method of peptide, be specifically related to a kind of production process of polypeptide that improves protein utilization.
Background technology
Polypeptide typically refers to the material in its peptide chain with 2-50 amino-acid residue.Compare with macromolecular protein, different polypeptide has different physiological functions, for example: delay the aging of body, reduce the generation of various geriatric diseases; Suppress skin aging; Hypotensive, reducing blood-fat, improves immunizing power.Polypeptide, except having various physiological functions, also can improve the stability of food, extends the shelf time of food, and have that solvability is good, the feature such as good processability, not irritated, easy absorption.Therefore, polypeptide is the important batching of food, healthcare products and nutritious prod.
The source of polypeptide comprises: extract from natural biological body (1); (2) in the process of pipe intestinal digesting protein, produce, or produce by external enzymolysis protein; (3) chemosynthesis, enzyme process synthesize or recombinant DNA technology is synthesized.Due to content seldom or extract difficulty, the polypeptide extracting from natural biological body can not meet the large-scale demand of the mankind.The chemical synthesis that pharmacology level polypeptide is conventional has side reaction to occur, and these by products are harmful.Production by Enzymes polypeptide, because it has working condition gentleness, the advantage such as safe, cheap, is the main stream approach of producing polypeptide.
The raw material of producing polypeptide comprises vegetable-protein, animal proteinum and algae protein.Vegetable-protein comprises soybean protein, wheat germ protein, zein, rape seed protein, Semen arachidis hypogaeae protein, tea seed albumen, sunflower protein etc.; Animal proteinum comprises milk-protein, fish protein, Protalbinic acid, collagen protein etc.; Algae protein comprises and splits kettle algae albumen, spirulina protein, Kou Shi Crypthecodinium cohnii albumen, my Ken Shi algae albumen etc.
The general combination that adopts a kind of proteolytic enzyme or multiple protein enzyme of existing production process of polypeptide, carries out primary enzymolysis, and its defect is that protein utilization is lower, generally below 40% (weight).In order to improve yield or protein utilization, conventionally take to increase enzyme consumption, extend the modes such as reaction times and meet, yet, in fact, the enzymolysis parameters such as the consumption of enzyme, reaction times are restrictions mutually, under certain conditions, by extending the reaction times, improve yield or protein utilization, its effect is also not obvious, and can produce a large amount of total free aminoacidss, affects the quality of product.
In addition, in the production process of polypeptide, one of problem facing is desalination, because repeatedly regulate pH value in enzymolysis process, in enzymolysis solution, the content of salt is higher.Although enzymolysis can make the hydrophobic amino acid that was originally imbedded in protein molecule inside come out, thereby make protein zymolyte from water, by hydrophobic interaction, be adsorbed onto the hydrophobic surface of resin, and can utilize the opposed polarity of different concns eluent to carry out grading purification to protein zymolyte, inorganic salt in zymolyte are not because being removed when the water elution by resin absorption, but these method complex process, cost is very high.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of production process of polypeptide that improves protein utilization, this production process of polypeptide is by three times relatively independent continuous enzymolysis, the protein utilization to raw material can be improved, colory polypeptide products can be obtained again.The technical scheme adopting is as follows:
Improve a production process of polypeptide for protein utilization, it is characterized in that in turn including the following steps:
(1) raw materials pretreatment: get proteinaceous raw material, obtain the first feed liquid after soaking;
(2) homogenization treatment: carry out homogenization treatment after the first feed liquid that step (1) is obtained stirs;
(3) Sumizyme MP enzymolysis: will adjust its temperature to 37-50 ℃ after the first feed liquid sterilizing after homogenization treatment, and regulate its pH value to 7.5-9.5, then add the proteolytic enzyme that accounts for raw material weight 1-5%, enzymolysis 1-3 hour in the situation that stirring;
This step proteolytic enzyme used is a kind of or wherein combination of two kinds in Sumizyme MP, trypsinase and Protamex compound protease;
(4) go out for the first time enzyme, centrifugation: the enzymolysis solution that step (3) the is obtained enzyme that goes out, then, with the centrifugal 10-15 minute of rotating speed of 3000-4500 rev/min, obtains supernatant A and deposit A;
(5) neutral protease enzymolysis: deposit A is added to water and stirs, obtain the second feed liquid; Then the temperature of the second feed liquid is adjusted to 37-50 ℃, and regulate its pH value to 7.0-7.5, then add the proteolytic enzyme that accounts for deposit A weight 2-8%, enzymolysis 1-3 hour in the situation that stirring;
This step proteolytic enzyme used is a kind of or wherein combination of two kinds in neutral protease, papoid and food flavor enzyme;
(6) go out for the second time enzyme, centrifugation: the enzymolysis solution that step (5) the is obtained enzyme that goes out, then, with the centrifugal 10-15 minute of rotating speed of 3000-4500 rev/min, obtains supernatant liquor B and deposit B;
(7) aspartic protease enzymolysis: deposit B is added to water and stirs, obtain the 3rd feed liquid; Then the temperature of the 3rd feed liquid is adjusted to 37-50 ℃, and regulate its pH value to 3-5, then add the proteolytic enzyme that accounts for deposit B weight 1-5%, enzymolysis 1-2 hour in the situation that stirring;
This step proteolytic enzyme used is a kind of in aspartic protease and stomach en-or both combinations;
(8) go out for the third time enzyme, centrifugation: the enzymolysis solution that step (7) the is obtained enzyme that goes out, then, with the centrifugal 10-15 minute of rotating speed of 3000-4500 rev/min, discard precipitation, obtain supernatant C;
(9) supernatant A, supernatant liquor B and supernatant C mixed and the pH value of its mixed solution is adjusted to 6.5-7.0, obtaining polypeptide mixed solution;
(10) remove the water in polypeptide mixed solution, obtain polypeptide.
In preferred steps (1), raw materials used protein content >35% (weight).
In preferred steps (1), the weight of the water adding is 8-10 times of raw material; Soak time is 30-60 minute.
In preferred steps (2), homogenization treatment is to carry out under 45-60 ℃, the pressure condition that is 10-20MPa in temperature.
In preferred steps (3), by the sterilizing 15-30 minute at 85-95 ℃ of the first feed liquid after homogenization treatment, be then cooled to 37-50 ℃.
In preferred steps (4), the enzymolysis solution that step (3) the is obtained enzyme 10-15 minute that goes out at 85-95 ℃.
In preferred steps (5), the weight of the water adding is 6-8 times of deposit A.
In preferred steps (6), the enzymolysis solution that step (5) the is obtained enzyme 10-15 minute that goes out at 85-95 ℃.
In preferred steps (7), the weight of the water adding is 6-8 times of deposit B.
In preferred steps (8), the enzymolysis solution that step (7) the is obtained enzyme 10-15 minute that goes out at 85-95 ℃.
In step (10), by spraying after polypeptide mixed solution vacuum concentration dry (vacuum concentration, dry being of spraying are removed the treating processes of anhydrating), obtain polypeptide.
The present invention is to forming the amino acid kind of substrate protein in raw material, quantity and sequence do not have special requirement, the raw material that can be used for producing polypeptide comprises the dregs of beans (containing vegetable-protein) after soybean degreasing, (Semen Brassicae campestris is carried the by product after oil to rapeseed meal, containing vegetable-protein), wheatgerm, peanut meal (the by product of Semen arachidis hypogaeae after oil plant is refined in squeezing, be rich in vegetable-protein), sunflower seed dregs (sunflower seeds squeezes the by product after grease through pre-pressing or direct leaching), splitting kettle algae carries and splits the kettle algae dregs of rice (containing algae protein) after oil, spirulina, Kou Shi Crypthecodinium cohnii or its are carried the algae dregs of rice (containing algae protein) after oil, my Ken Shi algae or its are carried the algae dregs of rice (containing algae protein) after oil, whey protein concentrate (containing animal proteinum), fish scale, pigskin etc.
The principle that the present invention is different to the enzymolysis site of protein according to different enzymes, adopt the mode of three enzymolysis, reach the object that improves protein utilization rate, that is: first under alkaline condition, adopt a kind of in Sumizyme MP, Protmex and trypsinase or wherein the combination of two kinds carry out enzymolysis; Under neutrallty condition, adopt afterwards a kind of in neutral protease, food flavor enzyme and papoid or wherein the combination of two kinds carry out enzymolysis; Finally under acidic conditions, adopt a kind of in aspartic protease and stomach en-or both combinations to carry out enzymolysis.By the enzymolysis under three different conditions, protein contained in raw material is fully used, protein utilization >80%, and the product content of peptides obtaining is high.
The present invention, by adopting suitable enzymolysis time, controls suitable Degree of Enzymatic Hydrolysis (degree of hydrolysis), to reduce the generation of total free aminoacids.And three times enzymolysis is relatively independent, after each enzymolysis, isolating supernatant liquor, then to precipitation continue enzymolysis (be to isolate supernatant liquor after Sumizyme MP enzymolysis, then with neutral protease the precipitation enzymolysis to its generation; After neutral protease enzymolysis, isolate supernatant liquor, then with aspartic protease the precipitation enzymolysis to its generation), further to reduce the generation of total free aminoacids, free aminoacid content in the finished product is reduced.
The present invention is suitably adjusting under the prerequisite of pH value, mode by reasonable controlled enzymatic hydrolysis time, reasonable controlled enzymatic hydrolysis order is controlled saltiness, during enzymolysis, only need adjust the initial pH value of every kind of enzymolysis solution, the product finally forming is recalled to neutral, without desalination, thereby simplify technique and reduce costs.
The polypeptide products good water solubility that the present invention obtains, more than protein content reaches 70% (weight), the batching that can be used as food, healthcare products and nutritious prod is used, in the product that is particularly suitable for solubleness and hypoallergenic to have relatively high expectations.
Embodiment
Embodiment 1
In the present embodiment, the production process of polypeptide that improves protein utilization in turn includes the following steps:
(1) raw materials pretreatment: get proteinaceous raw material (raw material is the dregs of beans after soybean degreasing, its protein content >45% (weight)), obtain the first feed liquid after soaking;
In this step (1), the weight of the water adding is 8 times of raw material; Soak time is 60 minutes.
(2) homogenization treatment: carry out homogenization treatment after the first feed liquid that step (1) is obtained stirs;
In this step (2), homogenization treatment is to carry out under 45 ℃, the pressure condition that is 20MPa in temperature.
(3) Sumizyme MP enzymolysis: will adjust its temperature to 37 ℃ after the first feed liquid sterilizing after homogenization treatment (by the sterilizing 30 minutes at 85 ℃ of the first feed liquid after homogenization treatment, then be cooled to 37 ℃), and regulate its pH value to 7.5, then add the proteolytic enzyme (being Sumizyme MP) that accounts for raw material weight 1%, in the situation that stirring, enzymolysis is 3 hours;
(4) go out for the first time enzyme, centrifugation: the enzymolysis solution that step (3) the is obtained enzyme (enzymolysis solution that step (3) is obtained go out at 85 ℃ enzyme 10 minutes) that goes out, then with the rotating speed of 3000 revs/min centrifugal 15 minutes, obtains supernatant A and deposit A;
(5) neutral protease enzymolysis: deposit A is added to water and stir (weight of the water adding is 6 times of deposit A), obtain the second feed liquid; Then the temperature of the second feed liquid is adjusted to 37 ℃, and regulate its pH value to 7.0, then add the proteolytic enzyme (being neutral protease) that accounts for deposit A weight 2%, in the situation that stirring, enzymolysis is 3 hours;
(6) go out for the second time enzyme, centrifugation: the enzymolysis solution that step (5) the is obtained enzyme (enzymolysis solution that step (5) is obtained go out at 95 ℃ enzyme 10 minutes) that goes out, then with the rotating speed of 3000 revs/min centrifugal 15 minutes, obtains supernatant liquor B and deposit B;
(7) aspartic protease enzymolysis: deposit B is added to water and stir (weight of the water adding is 6 times of deposit B), obtain the 3rd feed liquid; Then the temperature of the 3rd feed liquid is adjusted to 37 ℃, and regulate its pH value to 3, then add the proteolytic enzyme (being stomach en-) that accounts for deposit B weight 1%, in the situation that stirring, enzymolysis is 2 hours;
(8) go out for the third time enzyme, centrifugation: the enzymolysis solution that step (7) the is obtained enzyme (enzymolysis solution that step (7) is obtained go out at 85 ℃ enzyme 15 minutes) that goes out, then with the rotating speed of 3000 revs/min centrifugal 15 minutes, discard precipitation, obtain supernatant C;
(9) supernatant A, supernatant liquor B and supernatant C mixed and the pH value of its mixed solution is adjusted to 6.5, obtaining polypeptide mixed solution;
(10) remove the water in polypeptide mixed solution, obtain polypeptide.
In this step (10), by spraying after polypeptide mixed solution vacuum concentration dry (vacuum concentration, dry being of spraying are removed the treating processes of anhydrating), obtain polypeptide.
The protein utilization >80%(weight of the present embodiment), the protein content >70%(weight of the polypeptide products obtaining).
Embodiment 2
In the present embodiment, the production process of polypeptide that improves protein utilization in turn includes the following steps:
(1) raw materials pretreatment: get proteinaceous raw material (raw material is whey protein concentrate, its protein content >35% (weight)), obtain the first feed liquid after soaking;
In this step (1), the weight of the water adding is 10 times of raw material; Soak time is 30 minutes.
(2) homogenization treatment: carry out homogenization treatment after the first feed liquid that step (1) is obtained stirs;
In this step (2), homogenization treatment is to carry out under 60 ℃, the pressure condition that is 10MPa in temperature.
(3) Sumizyme MP enzymolysis: will adjust its temperature to 50 ℃ after the first feed liquid sterilizing after homogenization treatment (by the sterilizing 15 minutes at 95 ℃ of the first feed liquid after homogenization treatment, then be cooled to 50 ℃), and regulate its pH value to 9.5, then add the proteolytic enzyme (combination of trypsinase and Protamex compound protease that accounts for raw material weight 5%, wherein trypsinase and Protamex compound protease weight respectively account for half), in the situation that stirring, enzymolysis is 1 hour;
(4) go out for the first time enzyme, centrifugation: the enzymolysis solution that step (3) the is obtained enzyme (enzymolysis solution that step (3) is obtained go out at 95 ℃ enzyme 10 minutes) that goes out, then with the rotating speed of 4500 revs/min centrifugal 10 minutes, obtains supernatant A and deposit A;
(5) neutral protease enzymolysis: deposit A is added to water and stir (weight of the water adding is 8 times of deposit A), obtain the second feed liquid; Then the temperature of the second feed liquid is adjusted to 50 ℃, and regulate its pH value to 7.5, add the proteolytic enzyme (combination of papoid and food flavor enzyme, wherein papoid and food flavor enzyme weight respectively account for half) that accounts for deposit A weight 8%, in the situation that stirring, enzymolysis is 1 hour again;
(6) go out for the second time enzyme, centrifugation: the enzymolysis solution that step (5) the is obtained enzyme (enzymolysis solution that step (5) is obtained go out at 95 ℃ enzyme 10 minutes) that goes out, then with the rotating speed of 4500 revs/min centrifugal 10 minutes, obtains supernatant liquor B and deposit B;
(7) aspartic protease enzymolysis: deposit B is added to water and stir (weight of the water adding is 8 times of deposit B), obtain the 3rd feed liquid; Then the temperature of the 3rd feed liquid is adjusted to 50 ℃, and regulate its pH value to 5, then add the proteolytic enzyme (being stomach en-) that accounts for deposit B weight 5%, in the situation that stirring, enzymolysis is 1 hour;
(8) go out for the third time enzyme, centrifugation: the enzymolysis solution that step (7) the is obtained enzyme (enzymolysis solution that step (7) is obtained go out at 95 ℃ enzyme 10 minutes) that goes out, then with the rotating speed of 4500 revs/min centrifugal 10 minutes, discard precipitation, obtain supernatant C;
(9) supernatant A, supernatant liquor B and supernatant C mixed and the pH value of its mixed solution is adjusted to 7.0, obtaining polypeptide mixed solution;
(10) remove the water in polypeptide mixed solution, obtain polypeptide.
In this step (10), by spraying after polypeptide mixed solution vacuum concentration dry (vacuum concentration, dry being of spraying are removed the treating processes of anhydrating), obtain polypeptide.
The protein utilization >80%(weight of the present embodiment), the protein content >70%(weight of the polypeptide products obtaining).
Embodiment 3
In the present embodiment, the production process of polypeptide that improves protein utilization in turn includes the following steps:
(1) raw materials pretreatment: get proteinaceous raw material (raw material be split kettle algae carry after oil, split the kettle algae dregs of rice, its protein content >40% (weight)), after soaking, obtain the first feed liquid;
In this step (1), the weight of the water adding is 9 times of raw material; Soak time is 45 minutes.
(2) homogenization treatment: carry out homogenization treatment after the first feed liquid that step (1) is obtained stirs;
In this step (2), homogenization treatment is to carry out under 50 ℃, the pressure condition that is 15MPa in temperature.
(3) Sumizyme MP enzymolysis: will adjust its temperature to 45 ℃ after the first feed liquid sterilizing after homogenization treatment (by the sterilizing 20 minutes at 90 ℃ of the first feed liquid after homogenization treatment, then be cooled to 45 ℃), and regulate its pH value to 8.0, then add the proteolytic enzyme (being Protamex compound protease) that accounts for raw material weight 3%, in the situation that stirring, enzymolysis is 2 hours;
(4) go out for the first time enzyme, centrifugation: the enzymolysis solution that step (3) the is obtained enzyme (enzymolysis solution that step (3) is obtained go out at 90 ℃ enzyme 12 minutes) that goes out, then with the rotating speed of 4000 revs/min centrifugal 11 minutes, obtains supernatant A and deposit A;
(5) neutral protease enzymolysis: deposit A is added to water and stir (weight of the water adding is 7 times of deposit A), obtain the second feed liquid; Then the temperature of the second feed liquid is adjusted to 40 ℃, and regulate its pH value to 7.5, add the proteolytic enzyme (combination of papoid and food flavor enzyme, wherein papoid and food flavor enzyme weight respectively account for half) that accounts for deposit A weight 4%, in the situation that stirring, enzymolysis is 2 hours again;
(6) go out for the second time enzyme, centrifugation: the enzymolysis solution that step (5) the is obtained enzyme (enzymolysis solution that step (5) is obtained go out at 90 ℃ enzyme 12 minutes) that goes out, then with the rotating speed of 3500 revs/min centrifugal 15 minutes, obtains supernatant liquor B and deposit B;
(7) aspartic protease enzymolysis: deposit B is added to water and stir (weight of the water adding is 7 times of deposit B), obtain the 3rd feed liquid; Then the temperature of the 3rd feed liquid is adjusted to 45 ℃, and regulate its pH value to 4, add the proteolytic enzyme (aspartic protease and pepsic combination, wherein aspartic protease and stomach en-weight respectively account for half) that accounts for deposit B weight 3%, in the situation that stirring, enzymolysis is 1.5 hours again;
(8) go out for the third time enzyme, centrifugation: the enzymolysis solution that step (7) the is obtained enzyme (enzymolysis solution that step (7) is obtained go out at 90 ℃ enzyme 12 minutes) that goes out, then with the rotating speed of 3500 revs/min centrifugal 12 minutes, discard precipitation, obtain supernatant C;
(9) supernatant A, supernatant liquor B and supernatant C mixed and the pH value of its mixed solution is adjusted to 6.8, obtaining polypeptide mixed solution;
(10) remove the water in polypeptide mixed solution, obtain polypeptide.
In this step (10), by spraying after polypeptide mixed solution vacuum concentration dry (vacuum concentration, dry being of spraying are removed the treating processes of anhydrating), obtain polypeptide.
The protein utilization >80%(weight of the present embodiment), the protein content >70%(weight of the polypeptide products obtaining).
Claims (10)
1. improve a production process of polypeptide for protein utilization, it is characterized in that in turn including the following steps:
(1) raw materials pretreatment: get proteinaceous raw material, obtain the first feed liquid after soaking;
(2) homogenization treatment: carry out homogenization treatment after the first feed liquid that step (1) is obtained stirs;
(3) Sumizyme MP enzymolysis: will adjust its temperature to 37-50 ℃ after the first feed liquid sterilizing after homogenization treatment, and regulate its pH value to 7.5-9.5, then add the proteolytic enzyme that accounts for raw material weight 1-5%, enzymolysis 1-3 hour in the situation that stirring;
This step proteolytic enzyme used is a kind of or wherein combination of two kinds in Sumizyme MP, trypsinase and Protamex compound protease;
(4) go out for the first time enzyme, centrifugation: the enzymolysis solution that step (3) the is obtained enzyme that goes out, then, with the centrifugal 10-15 minute of rotating speed of 3000-4500 rev/min, obtains supernatant A and deposit A;
(5) neutral protease enzymolysis: deposit A is added to water and stirs, obtain the second feed liquid; Then the temperature of the second feed liquid is adjusted to 37-50 ℃, and regulate its pH value to 7.0-7.5, then add the proteolytic enzyme that accounts for deposit A weight 2-8%, enzymolysis 1-3 hour in the situation that stirring;
This step proteolytic enzyme used is a kind of or wherein combination of two kinds in neutral protease, papoid and food flavor enzyme;
(6) go out for the second time enzyme, centrifugation: the enzymolysis solution that step (5) the is obtained enzyme that goes out, then, with the centrifugal 10-15 minute of rotating speed of 3000-4500 rev/min, obtains supernatant liquor B and deposit B;
(7) aspartic protease enzymolysis: deposit B is added to water and stirs, obtain the 3rd feed liquid; Then the temperature of the 3rd feed liquid is adjusted to 37-50 ℃, and regulate its pH value to 3-5, then add the proteolytic enzyme that accounts for deposit B weight 1-5%, enzymolysis 1-2 hour in the situation that stirring;
This step proteolytic enzyme used is a kind of in aspartic protease and stomach en-or both combinations;
(8) go out for the third time enzyme, centrifugation: the enzymolysis solution that step (7) the is obtained enzyme that goes out, then, with the centrifugal 10-15 minute of rotating speed of 3000-4500 rev/min, discard precipitation, obtain supernatant C;
(9) supernatant A, supernatant liquor B and supernatant C mixed and the pH value of its mixed solution is adjusted to 6.5-7.0, obtaining polypeptide mixed solution;
(10) remove the water in polypeptide mixed solution, obtain polypeptide.
2. production process of polypeptide according to claim 1, is characterized in that: in step (1), and raw materials used protein content >35% (weight).
3. production process of polypeptide according to claim 1, is characterized in that: in step (1), the weight of the water adding is 8-10 times of raw material; Soak time is 30-60 minute.
4. production process of polypeptide according to claim 1, is characterized in that: in step (2), homogenization treatment is to carry out under 45-60 ℃, the pressure condition that is 10-20MPa in temperature.
5. production process of polypeptide according to claim 1, is characterized in that: in step (3), by the sterilizing 15-30 minute at 85-95 ℃ of the first feed liquid after homogenization treatment, be then cooled to 37-50 ℃.
6. production process of polypeptide according to claim 1, is characterized in that: in step (4), and the enzymolysis solution that step (3) the is obtained enzyme 10-15 minute that goes out at 85-95 ℃.
7. production process of polypeptide according to claim 1, is characterized in that: in step (5), the weight of the water adding is 6-8 times of deposit A.
8. production process of polypeptide according to claim 1, is characterized in that: in step (6), and the enzymolysis solution that step (5) the is obtained enzyme 10-15 minute that goes out at 85-95 ℃.
9. production process of polypeptide according to claim 1, is characterized in that: in step (7), the weight of the water adding is 6-8 times of deposit B.
10. production process of polypeptide according to claim 1, is characterized in that: in step (8), and the enzymolysis solution that step (7) the is obtained enzyme 10-15 minute that goes out at 85-95 ℃.
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