CN100392092C - Soybean peptide production method - Google Patents
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Abstract
The present invention relates to a production method for soybean peptides. In the production method, degreased bean pulp or / and soybean flour can be used as raw materials. The present invention comprises the process steps: 1) solid fermentation: raw materials and water are mixed and are uniformly stirred according to a certain weight proportion, and then zymocyte is inoculated for fermentation by controlling temperature; 2) liquid enzymolysis: fermented and cultured solid substances and water are mixed according to a certain proportion, composite proteinases are added for enzymolysis by heat insulation; 3) enzyme sterilization: enzymes are sterilized with degradable substances; 4) liquid filtration: a crude soybean peptide solution is separated from an enzymolysis liquid; 5) debitterization and decolorization; 6) refining; 7) sterilization to obtain finished products. The present invention has the advantage that the present invention combines the advantages of a liquid fermentation method and a liquid enzymolysis method, overcomes the defects thereof, lowers material costs, greatly reduces external enzyme quantity, obtains high hydrolysis ratio and has an important promotion action for the popularization and the application of soybean peptides.
Description
Technical field
The present invention relates to soybean peptide production method, belong to the technical field that the protein molecule in microbiology, the zymetology is transformed.
Background technology
Soybean peptides is the deep processed product of soybean, mainly is to utilize biological method, and the soybean transformation protein molecule is low molecular peptide, its physiology and trophic function are significantly improved and improves.Compare with traditional soybean albumen, the production technique of soybean peptides is complicated, cost value is higher, digest and assimilate, can supply with human body energy, no protein denaturation, free from beany flavor rapidly but it has easily, no residue, liquid viscosity is little and be heated and characteristic such as do not solidify, and especially has the serum cholesterol of reduction, hypotensive and promote special physiological properties such as metabolism of fat.Therefore soybean peptides is than soybean protein more fine, novel soybean deep processed product and nutritious prod, has demonstrated tempting development prospect in fields such as food, medicine, daily-use chemical industry.For example, soy-bean peptide can promote the secretion of albumen synthetic hormone, can carry out effective for repairing for the muscle of being damaged in the match; Show in the effect evaluation and test that many tame professional sports teams carry out: drink continuously, can obviously strengthen athletic muscle power and endurance, and can make muscle fatigue eliminate rapidly, regain one's strength.
Soybean peptides is except that having multiple nutrients is worth the effect that also have reducing cholesterol, brings high blood pressure down.With soybean peptides can produce decreasing cholesterol, hypotensive and the prevention cardiovascular system diseases protective foods.Soybean peptides can directly be absorbed by enteron aisle, under special physical qualification, can satisfy the requirement of keeping and improving the proteinaceous nutrient state, particularly, can be used as intestinal feeding agent and fluidised form food and directly eat for post-operative recovery patient, infant, man at an advanced age etc.In addition, nonspecific effect that soybean peptides can also play and build up health, add the powerful antibody defense mechanism, improve quality of life, delays senility, this is undoubtedly the most useful a kind of compensation for alleviating human body because of can't self producing the relevant composition shortage present situation that the enough various functional peptides of function causes.
Because the effect and the application prospect of soybean peptides have been carried out number of research projects both at home and abroad.From the method characteristics, all be one-step method production process, or the method for liquid fermenting, or the method for liquid enzymolysis.
The method production soybean peptides of liquid fermenting is that soyflour or soybean protein isolate are added the water mixing earlier, after the high-temperature steam sterilization, inoculates specific microorganism strains and ferments.The microorganism type of inoculation is different and variant according to data source, and Mucor, aspergillus oryzae, aspergillus niger, bacillus natto, subtilis etc. are arranged, and only inoculation a kind of microorganism is wherein fermented usually.Behind the fermentation certain hour, by separation, concentrated and dry, make the soybean peptides product again.Liquid fermentation process production soybean peptides cost height, there is fluctuation in quality product.The problem of several respects below main the existence: 1, need high-temperature sterilization and airtight condition, investment is big; 2, the conditional request of the growth conditions of thalline and enzymolysis is variant, can not satisfy both requirements in the production simultaneously.Because in the fermenting process, bacterium produces enzyme, and the enzyme decomposing protein is to play a role simultaneously substantially.The temperature that enzymolysis requires will be higher than the growth temperature requirement of bacterium usually.Satisfying enzymatic hydrolysis condition, will certainly influence the growth of bacterium, also is like this conversely.Enzyme derives from bacterium, so can only control by the growth temperature of bacterium on producing.Therefore, production efficiency is affected, and the product yield is low.3, fermentation produces bacterium earlier, produces enzyme again, passes through the effect hydrolytic soya bean protein of enzyme then.Because amount and enzymatic hydrolysis condition to the enzyme that produces can't effectively be controlled, and tackle proteinic hydrolysis degree mutually and can not accurately control.Therefore, the stabilised quality of soybean peptides product is difficult to be protected.Just because of these deficiencies, liquid fermentation process production soybean peptides does not form suitability for industrialized production.
It also is earlier soyflour or soybean protein isolate to be added the water mixing that the method for liquid enzymolysis is produced soybean polypeptide, directly adds one or more compound proteases then and is incubated hydrolysis.When utilizing the conjugated protein enzymic hydrolysis, some investigator adopts the mode that once adds, and some is the mode that adopts secondary to add, and in enzymolysis process, adds different enzymes in different time sections.The neutral protease in the aspartic protease in the proteolytic enzyme in the mould source of lytic enzyme type hairiness of using, aspergillus niger source, aspergillus oryzae source, the proteolytic enzyme (Alcalase, Alcakase, Protamex etc.) of bacterial origin, the proteolytic enzyme of plant origin (papoid, bromeline etc.).After hydrolysis is finished, more successively by heating go out enzyme, cooling, separation, concentrate and dry, make the soybean peptides product at last.Liquid enzymes solution production soybean peptides working condition is relatively gentleer, does not need high-temperature high-pressure apparatus, compares with fermentation method, and investment reduces relatively.Secondly, liquid enzymes solution production soybean peptides can realize more accurate control to the amount and the enzymatic hydrolysis condition of enzyme, and quality product is more stable.Liquid enzymes solution production soybean peptides has entered suitability for industrialized production, but liquid enzymes solution production soybean peptides still cost is higher, limited the application of soybean peptides product.The higher reason one of cost is that the liquid enzymes solution is low to low price coarse raw materials hydrolysis yields such as soyflours, therefore use the raw material of high purity high price, the general soybean protein isolate that adopts is as raw materials for production, and equal protein raw material cost has increased more than 5 times than soyflour; The 2nd, the enzyme amount that the liquid enzymes solution is used is big, the cost height of outsourcing enzyme.
Summary of the invention
The objective of the invention is to: the present invention has absorbed the advantage of solution fermentation and liquid enzymes solution, has overcome their deficiency, and a kind of method production soybean peptides that adopts two-stage method, promptly combines by solid fermentation and liquid enzymolysis is provided.2) and can finish thereby 1), can be: to soy proteinaceous prehydrolysis with defatted soybean meal or/and soyflour, reduces raw materials cost as raw material; 3) thereby, had solid fermentation to make the amount that can reduce additional enzyme in the enzymic hydrolysis stage in a large number, reduce cost effectively and obtain higher percent hydrolysis.
Technical scheme of the present invention is: described soybean peptide production method, it is with defatted soybean meal or/and soyflour is a raw material, 1), solid fermentation its processing step comprises:: with raw material and water is 1: 0.5～1.5 to mix and mix all by weight, inoculation fermentation bacterium subsequently, temperature controlled fermentation; 2), liquid enzymolysis: solid culture that will be by fermentation and water are 1: 7～10 to mix by weight, and the adding compound protease is incubated enzymolysis; 3), the enzyme that goes out: the degradation product enzyme that goes out; 4), filtrate: enzymolysis solution is isolated the thick solution of soybean peptides; 5), debitterizing and decoloring; 6), refining, 7), required finished product is made in sterilization.
The zymophyte that above-mentioned solid fermentation is inoculated is a subtilis, inoculative proportion 0.1～5%, and the control leavening temperature is 25～50 ℃, feed height 50～90cm, fermentation time are 1.5～2.5 days.
The compound protease that the aforesaid liquid enzymolysis is added is by fermentation solid culture to be added water mix the back and add the plant-sourced compound protease by 0.05～0.15%; The insulation enzymolysis is 45～60 ℃ of insulation hydrolysis 1.5～13 hours, and enzymolysis process slowly stirs continuously.
Raw material and water weight ratio are 1: 0.8～1.2 in the above-mentioned solid fermentation; Solid culture and water weight ratio are 1: 8.5～9.5 after the fermentation of described liquid enzymolysis.Be preferably: raw material and water weight ratio are 1: 1 in the solid fermentation; Solid culture and water weight ratio are 1: 9 after the fermentation of described liquid enzymolysis.
The inoculative proportion of above-mentioned inoculation subtilis is 4～5%, and the control leavening temperature is 28～45 ℃, and feed height 60～80cm, fermentation time are 1.5～2 days.
By fermentation solid culture adds water to mix the amount that the back adds the plant-sourced compound protease is 0.08～0.1% in the aforesaid liquid enzymolysis; Described insulation enzymolysis is 50～55 ℃ of insulation hydrolysis 2～10 hours.
The above-mentioned enzyme that goes out is after enzymolysis finishes, and enzymolysis solution is warming up to 70～85 ℃, keeps 10～30 minutes; Described debitterizing and decoloring is the filtrate operation to be separated the thick solution of the resulting soybean peptides in back be warming up to 50～65 ℃, by 0.05～0.15% adding gac, slowly stirs 0.5-2.5 hour, carries out debitterizing and decoloring and handles.
Above-mentioned refining is to adopt whizzer to separate earlier, gets soybean peptides solution clearly, carries out fractional separation with ultra-filtration membrane by molecular weight again, and molecular weight cut-off is respectively: 200,1000,5000 and 10000; Described sterilization is that filtrate is passed through 4-8 high-temperature instantaneous sterilizations in second at 130～140 ℃, obtains sterilization soybean peptides solution-type product.
Above-mentioned formulation product is a pulvis, 1), concentrate method steps comprises:: the sterilization soybean peptides solution after the sterilization is placed the rotary evaporation in vacuo device, be decompressed to-0.8Mpa～-0.9Mpa, evaporation concentration under 65～75 ℃ of constant temperatures reaches 15～30% to solid content and finishes; 2), spraying drying: under 0.08～0.15Mpa pressure condition, 102～110 ℃ of temperature, 75～90 ℃ of temperature outs are carried out spraying drying in 130～140 ℃ of inlet temperatures, the tower, finally obtain the powdery soybean peptide product with concentrated solution.
The advantage of soybean peptide production method of the present invention is: a kind of method production soybean peptides that adopts two-stage method, promptly combines by solid fermentation and liquid enzymolysis is provided.1), the method that combines of solid fermentation method and solid fermentation and liquid enzymolysis, be a kind of new producing and manufacturing technique, obviously distinguish the soybean peptide production method of single liquid fermenting and single liquid enzymolysis in the prior art.Solid fermentation is by specific bacterial strain raw material to be carried out solid fermentation, accumulates a large amount of cytohydrolists and proteolytic ferment, thereby can be with using defatted soybean meal or/and soyflour as raw material, reduces raw materials cost; 2) and can finish to soy proteinaceous prehydrolysis; 3) thereby, had solid fermentation to make can to reduce the amount of additional enzyme in a large number in the enzymic hydrolysis stage, and obtain higher percent hydrolysis.The present invention has absorbed the advantage of solution fermentation and liquid enzymes solution, has overcome their deficiency, and the ton production cost descends more than one times, and the penetration and promotion of soybean peptides is used great promoter action.
Embodiment
Below enumerate embodiment and further explain the present invention, but the present invention is not subjected to the restriction of these specific embodiments.
Embodiment 1: be raw material with the soyflour, when being that to add soyflour be that raw material belongs to additional embodiments for raw material or defatted soybean meal with the defatted soybean meal; Its processing step is:
1), solid fermentation: with soyflour is that raw material is 1: 1 with water for 300 kilograms by weight, i.e. 300 kg of water mixing is mixed all, and Jie Zhong zymophyte is a subtilis subsequently, inoculative proportion 5%, the control leavening temperature is 28-45 ℃, and feed height 50cm, fermentation time are 2 days;
2), liquid enzymolysis: solid culture that will be by fermentation and water are to mix at 1: 9 by weight, and the adding compound protease is incubated enzymolysis; The compound protease that is added is by fermentation solid culture to be added water mix the back and add the plant-sourced compound protease by 0.1%; The insulation enzymolysis is 55 ℃ of insulation hydrolysis 9 hours, and enzymolysis process slowly stirs continuously;
3), enzyme goes out: after enzymolysis finishes, enzymolysis solution is warming up to 80 ℃, kept 15 minutes;
4), filtrate: adopt filter press to separate enzymolysis solution, the solution that press filtration goes out is the thick solution of soybean peptides;
5), debitterizing and decoloring, be the filtrate operation to be separated the thick solution of the resulting soybean peptides in back be warming up to 60 ℃, add gac by 0.1%, slowly stirred 1.5 hours, carry out debitterizing and decoloring and handle;
6), refining, be to adopt earlier the 20000rpm supercentrifuge to separate, soybean peptides solution clearly, carry out fractional separation with ultra-filtration membrane by molecular weight again, molecular weight cut-off is respectively: 200Da, 1000Da, 5000Da and 10000Da;
7), sterilization makes finished product, be with filtrate at 135 ℃, through 5 second high-temperature instantaneous sterilization, obtain sterilization soybean peptides solution-type product; 1), concentrate continue to make required powder product, its method is:: the sterilization soybean peptides solution after the sterilization is placed the rotary evaporation in vacuo device, be decompressed to-0.9Mpa, evaporation concentration under 70 ℃ of constant temperatures reaches 20% to solid content and finishes; 2), spraying drying: under the 0.1Mpa pressure condition, 105 ℃ of temperature, 80 ℃ of temperature outs are carried out spraying drying in 135 ℃ of inlet temperatures, the tower, finally obtain the powdery soybean peptide product with concentrated solution.
Embodiment 2: its processing step is:
1), solid fermentation: be 600 kilograms of raw materials with water with soyflour be to mix at 1: 0.8 to mix all by weight, inoculate subtilis by 3% ratio, leavening temperature is 25-50 ℃, and feed height 90cm, fermentation time are 1.5 days;
2), the liquid enzymolysis: solid culture that will be by fermentation and water are to mix at 1: 7 by weight, add compound protease and be by fermentation solid culture added water and mixes after by 0.05% adding plant-sourced compound protease; 45 ℃ are incubated hydrolysis 13 hours, and enzymolysis process slowly stirs continuously;
3), enzyme goes out: enzymolysis solution is warming up to 70 ℃, kept 30 minutes;
4), filtrate: adopt centrifugal separation method to obtain the thick solution of soybean peptides enzymolysis solution;
5), debitterizing and decoloring, be the filtrate operation to be separated the thick solution of the resulting soybean peptides in back be warming up to 50 ℃, add gac by 0.05%, slowly stirred 0.5 hour, carry out debitterizing and decoloring and handle;
6), refining, be to adopt earlier the 20000rpm supercentrifuge to separate, soybean peptides solution clearly, carry out fractional separation with ultra-filtration membrane by molecular weight again, molecular weight cut-off is respectively: 200Da, 1000Da, 5000Da and 10000Da;
7), sterilization makes finished product, be with filtrate at 130 ℃, through 4 second high-temperature instantaneous sterilization, obtain sterilization soybean peptides solution-type product; 1), concentrate continue to make required powder product, its method is:: the sterilization soybean peptides solution after the sterilization is placed the rotary evaporation in vacuo device, be decompressed to-0.8Mpa, evaporation concentration under 65 ℃ of constant temperatures reaches 15% to solid content and finishes; 2), spraying drying: under the 0.08Mpa pressure condition, 102 ℃ of temperature, 75 ℃ of temperature outs are carried out spraying drying in 130 ℃ of inlet temperatures, the tower, finally obtain the powdery soybean peptide product with concentrated solution.
Embodiment 3: its processing step is:
1), solid fermentation: with 400 kilograms of defatted soybean meal raw materials and water is to mix at 1: 1.5 to mix all by weight, inoculates subtilis by 0.1% ratio, and leavening temperature is 30-45 ℃, and feed height 40-70cm, fermentation time are 2.5 days;
2), the liquid enzymolysis: solid culture that will be by fermentation and water are to mix at 1: 10 by weight, add compound protease and be by fermentation solid culture added water and mixes after by 0.15% adding plant-sourced compound protease; 60 ℃ are incubated hydrolysis 1.5 hours, and enzymolysis process slowly stirs continuously;
3), enzyme goes out: enzymolysis solution is warming up to 85 ℃, kept 10 minutes;
4), filtrate: adopt centrifugal separation method to obtain the thick solution of soybean peptides enzymolysis solution;
5), debitterizing and decoloring, be the filtrate operation to be separated the thick solution of the resulting soybean peptides in back be warming up to 65 ℃, add gac by 0.15%, slowly stirred 2.5 hours, carry out debitterizing and decoloring and handle;
6), refining, be to adopt earlier the 20000rpm supercentrifuge to separate, soybean peptides solution clearly, carry out fractional separation with ultra-filtration membrane by molecular weight again, molecular weight cut-off is respectively: 200Da, 1000Da, 5000Da and 10000Da;
7), sterilization makes finished product, be with filtrate at 140 ℃, through 8 second high-temperature instantaneous sterilization, obtain sterilization soybean peptides solution-type product; 1), concentrate continue to make required powder product, its method is:: the sterilization soybean peptides solution after the sterilization is placed the rotary evaporation in vacuo device, be decompressed to-0.9Mpa, evaporation concentration under 75 ℃ of constant temperatures reaches 30% to solid content and finishes; 2), spraying drying: under the 0.15Mpa pressure condition, 110 ℃ of temperature, 90 ℃ of temperature outs are carried out spraying drying in 140 ℃ of inlet temperatures, the tower, finally obtain the powdery soybean peptide product with concentrated solution.
Embodiment 4: different with the foregoing description is that the parameter range of choice is:
Raw material and water weight ratio are 1: 0.8～1.2 in the solid fermentation; Solid culture and water weight ratio are 1: 8.5～9.5 after the fermentation of described liquid enzymolysis;
The inoculative proportion of inoculation subtilis is 4～5%, and the control leavening temperature is 28～45 ℃, and feed height 60～80cm, fermentation time are 1.5～2 days;
It is 0.08～0.1% that solid culture by fermentation adds the amount that adds the plant-sourced compound protease after water mixes; Described insulation enzymolysis is 50～55 ℃ of insulation hydrolysis 2～10 hours.
Claims (9)
1), solid fermentation 1. a soybean peptide production method is characterized in that, or/and soyflour is a raw material, its processing step comprises with defatted soybean meal:: with raw material and water is 1: 0.5～1.5 to mix and mix all by weight, inoculation fermentation bacterium subsequently, temperature controlled fermentation; 2), liquid enzymolysis: solid culture that will be by fermentation and water are 1: 7～10 to mix by weight, and the adding compound protease is incubated enzymolysis; 3), the enzyme that goes out: the degradation product enzyme that goes out; 4), filtrate: enzymolysis solution is isolated the thick solution of soybean peptides; 5), debitterizing and decoloring; 6), refining; 7), required finished product is made in sterilization; The zymophyte that described solid fermentation is inoculated is a subtilis, inoculative proportion 0.1～5%, and the control leavening temperature is 25～50 ℃, feed height 50～90cm, fermentation time are 1.5～2.5 days; Described debitterizing and decoloring is the filtrate operation to be separated the thick solution of the resulting soybean peptides in back be warming up to 50～65 ℃, by 0.05～0.15% adding gac, slowly stirs 0.5-2.5 hour, carries out debitterizing and decoloring and handles.
2. soybean peptide production method according to claim 1 is characterized in that: the compound protease that the liquid enzymolysis is added is by fermentation solid culture to be added water mix the back and add the plant-sourced compound protease by 0.05～0.15%; The insulation enzymolysis is 45～60 ℃ of insulation hydrolysis 1.5～13 hours, and enzymolysis process slowly stirs continuously.
3. soybean peptide production method according to claim 1 is characterized in that: raw material and water weight ratio are 1: 0.8～1.2 in the described solid fermentation; Solid culture and water weight ratio are 1: 8.5～9.5 after the fermentation of described liquid enzymolysis.
4. soybean peptide production method according to claim 1 is characterized in that: raw material and water weight ratio are 1: 1 in the described solid fermentation; Solid culture and water weight ratio are 1: 9 after the fermentation of described liquid enzymolysis.
5. soybean peptide production method according to claim 1 is characterized in that: the inoculative proportion of inoculation subtilis is 4～5%, and the control leavening temperature is 28～45 ℃, and feed height 60～80cm, fermentation time are 1.5～2 days.
6. soybean peptide production method according to claim 2 is characterized in that: it is 0.08～0.1% that solid culture by fermentation adds the amount that adds the plant-sourced compound protease after water mixes; Described insulation enzymolysis is 50～55 ℃ of insulation hydrolysis 2～10 hours.
7. soybean peptide production method according to claim 1 is characterized in that: the described enzyme that goes out is after enzymolysis finishes, and enzymolysis solution is warming up to 70～85 ℃, keeps 10～30 minutes.
8. soybean peptide production method according to claim 1, it is characterized in that: described refining is to adopt whizzer to separate earlier, get soybean peptides solution clearly, carry out fractional separation with ultra-filtration membrane by molecular weight again, molecular weight cut-off is respectively: 200Da, 1000Da, 5000Da and 10000Da; Described sterilization is that filtrate is passed through 4-8 high-temperature instantaneous sterilizations in second at 130～140 ℃, obtains sterilization soybean peptides solution-type finished product.
9. according to the soybean peptide production method of claim 1, it is characterized in that: described finished product is a pulvis, 1), concentrate method steps comprises:: the sterilization soybean peptides solution after the sterilization is placed the rotary evaporation in vacuo device, be decompressed to-0.8Mpa～-0.9Mpa, evaporation concentration under 65～75 ℃ of constant temperatures reaches 15～30% to solid content and finishes; 2), spraying drying: under 0.08～0.15Mpa pressure condition, 102～110 ℃ of temperature, 75～90 ℃ of temperature outs are carried out spraying drying in 130～140 ℃ of inlet temperatures, the tower, finally obtain powdery soybean peptide finished product with concentrated solution.
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| CN103045705A (en) * | 2012-12-14 | 2013-04-17 | 广东海洋大学 | Method for producing protein peptide composed of fish protein peptide and soybean peptide |
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| CN110623137A (en) * | 2019-10-30 | 2019-12-31 | 创客商盟有限公司 | Soybean peptide product with small molecular weight unit and preparation method thereof |
| CN111631266A (en) * | 2020-06-04 | 2020-09-08 | 龙王致新(北京)科技有限公司 | Preparation method of peptide-containing high-protein soybean milk powder |
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| CN113981030A (en) * | 2021-12-10 | 2022-01-28 | 厦门元之道生物科技有限公司 | Method for preparing soybean oligopeptide by combining enzyme method with microbial fermentation and application thereof |
| CN114395600B (en) * | 2022-01-14 | 2023-12-12 | 琛蓝(美国)营养制品股份有限公司 | Preparation method and application of multifunctional pea peptide |
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