CN100338224C - Method for preparing peptide of soybean through continuous enzyme membrane reaction of soyabean protein - Google Patents
Method for preparing peptide of soybean through continuous enzyme membrane reaction of soyabean protein Download PDFInfo
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- CN100338224C CN100338224C CNB2004100129663A CN200410012966A CN100338224C CN 100338224 C CN100338224 C CN 100338224C CN B2004100129663 A CNB2004100129663 A CN B2004100129663A CN 200410012966 A CN200410012966 A CN 200410012966A CN 100338224 C CN100338224 C CN 100338224C
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Abstract
The present invention relates to a method for preparing soybean peptides by a soybean protein continuous enzyme membrane reaction, which carries out a soybean protein enzyme membrane reaction in an ultrafiltrate separation process and carries out activated carbon adsorption and soybean peptide concentration in a nanofiltration separation process. The present invention can simultaneously realize the cyclic usage of an enzyme preparation and a mother liquid and the processes of decoloration, debitterization and thickening for soybean peptides. The present invention not only can increase reaction conversion rate and product quality, but also can avoid waste liquid processing problem caused by the large consumption of the mother liquid and can realize continuous reaction and enclosed operation.
Description
Technical field
The invention belongs to the method field of soybean protein being carried out continuous enzyme membrane prepared in reaction soybean peptides.
Background technology
Soybean peptides is by soybean protein or the defatted soybean meal low-peptide mixer with extensive molecular weight distribution through the protease hydrolyzed gained.Soybean peptides can promote milk-acid bacteria, the propagation of bifidus bacillus and the secretion of useful metabolite, and solvability is not subjected to the influence of pH and heating, can be widely used in the exploitation of protective foods and functional food.But soybean protein often produces a kind of unacceptable bitter taste behind enzymolysis, need carry out suitable processing.The different soybean peptides of molecular weight has different physiologically actives, and the excessive or too small soybean peptides activity of molecular weight is all not high, and molecular weight that it is generally acknowledged the soybean peptides with physiologically active is mostly in 300～5000 scope; Therefore, the control of molecular weight directly has influence on the performance of soybean peptides product.The traditional method of preparation soybean peptides generally comprises a plurality of steps: soybean protein → enzyme digestion reaction → sterilization → centrifugation → supernatant liquor decolouring debitterize → gel filtration chromatography or gradocol membrane separation → vacuum concentration and drying, the range of molecular weight distributions of gained soybean peptides is bigger, and molecular weight reaches more than 20% greater than 5000 soybean peptides proportion.
Enzyme membrane reaction (EMR) is typical biocatalysis film reaction process, and biological degradation is to use enzyme membrane to react extensive fields the most.The enzyme membrane reaction process is three kinds of function integrations such as separating of the repeated use of the katalysis of enzyme, enzyme and product, thereby simplifies biodegradable technical process and operation steps.The method of existing employing enzyme membrane prepared in reaction soybean peptides can be controlled the range of molecular weight distributions of product effectively, improves product activity.Yet, in the enzyme membrane reaction process, when removing the enzymolysis product of lower molecular weight, also mother liquor in the reaction system (by preparations such as water and small molecules acid or alkali) can be removed, therefore, carrying out along with the enzyme membrane reaction, the solid content and the viscosity of trapped fluid (bio-degradation reactions substrate) obviously increase, the pH value produces fluctuation, restricted the activity of enzyme, thereby made enzymolysis be difficult to realize continuously, directly influenced soybean protein and prepared the transformation efficiency of soybean peptides and the raising of quality product, productivity can't improve simultaneously, and production cost can fall yet.
Summary of the invention
The technical problem to be solved in the present invention provides the method for the continuous enzyme membrane prepared in reaction of a kind of soybean protein soybean peptides.
The present invention solves the problems of the technologies described above with following technical scheme: the soybean peptides solution that obtains in the ultra-filtration and separation process directly enters nanofiltration system, the bigger soybean peptides of soybean protein and molecular weight is rested on continue enzymolysis in the trapped fluid, and the soybean peptides that meets the molecular weight distribution requirement is carried out the activated carbon decolorizing debitterize as stock liquid and is concentrated in nanofiltration system, and the liquid that sees through of nanofiltration process is sent into the recycling of enzyme digestion reaction system again to keep carrying out continuously of enzymolysis.
The method of the continuous enzyme membrane prepared in reaction of soybean protein of the present invention soybean peptides, with ultra-filtration technique and enzyme digestion reaction coupling, nanofiltration and charcoal absorption coupling, reclaim mother liquor and pump into the fermentor tank recycle by nanofiltration system, to keep the stable of reaction system concentration, viscosity and pH value, can just hydrolysate of soybean protein be more abundant in continuous enzyme membrane reaction, obtain higher transformation efficiency, molecular weight is higher less than 5000 ratio in the product.The present invention can realize simultaneously to zymin and mother liquor recycle and reclaim and to the decolouring debitterize and the concentration of soybean peptides.Can improve reaction conversion ratio and product quality, can avoid again producing the problem that causes producing a large amount of waste liquids because of a large amount of consumption mother liquors, realize continuous reaction, closed operation.
Description of drawings
Fig. 1 is the method technological process block-diagram of the continuous enzyme membrane prepared in reaction of soybean protein of the present invention soybean peptides.
Embodiment
The method main technique step of the continuous enzyme membrane prepared in reaction of soybean protein of the present invention soybean peptides is:
(1) carries out reacting based on the soybean protein enzyme membrane of ultra-filtration and separation;
(2) carry out concentrating based on the charcoal absorption and the soybean peptides of nanofiltration separation;
(3) drying of soybean peptides.
Method of the present invention mainly is to hold back by the selectivity of ultra-filtration membrane, the bigger soybean peptides of soybean protein and molecular weight is stayed in trapped fluid, carry out secondary enzymolysis, carry out the activated carbon decolorizing debitterize and concentrate in nanofiltration system as seeing through liquid and meet soybean peptides that molecular weight distribution requires, the liquid (mother liquor) that sees through of nanofiltration process is sent into the recycling of enzyme digestion reaction system again.In the recycling zymin, mother liquor is reclaimed and recycles, solve that loss owing to mother liquor causes enzyme to be suffered a loss, reaction conversion ratio is low, the serialization degree is low, production concentration and purity high-technology problem not, improve reaction conversion ratio and enzymolysis product quality, and reduced the discharging of waste liquid and the energy consumption of drying treatment.
The method of the continuous enzyme membrane prepared in reaction of soybean protein of the present invention soybean peptides, can adopt following steps and processing condition to carry out:
The first step is reacted based on the enzyme membrane of ultra-filtration and separation:
Soybean protein is dissolved in a certain amount of deionized water or the distilled water, and be that benchmark adds zymin by a certain percentage with the quantity of soybean protein, mechanical stirring is mixed into reaction raw materials liquid, and, carry out the enzyme membrane reaction with the pH value of certain density acid solution or alkali lye conditioned reaction system.The trapped fluid of ultra-filtration process is sent back to and is continued to serve as reaction substrate in the retort, is used to carry out the concentrated and charcoal absorption processing of nanofiltration and see through liquid.Nanofiltration concentrates and charcoal absorption is handled when concentration ratio (i.e. the volume ratio of reaction raw materials liquid before and after the reaction) begins when reaching certain value to carry out.
The soybean protein addition be deionized water or the distillation water yield 3%～10% (W/V), the enzyme addition with enzyme activity to soybean protein quality 1.0 * 10
-2AU/g～3.0 * 10
-2AU/g is a standard, and zymin can be selected Sumizyme MP for use, and accent pH value of reaction system such as neutral protease or aspartic protease are 3.0～10.0,50 ℃～85 ℃ of temperature of reaction; Rotating speed is chosen as 10r/min～100r/min during stirring, stirs back unlatching ultrafiltration system and carries out.The molecular weight cut-off 3000～5000 of ultra-filtration membrane; Membrane filtration pressure 0.08mPa～0.30mPa; Reaction times 4h～8h; Beginning to carry out nanofiltration when concentration ratio reaches 1.1～2.0 handles.
Charcoal absorption and the product of second step based on nanofiltration separation concentrates
The liquid pump that sees through of nanofiltration is got back to retort and is reused, and trapped fluid is flowed through after gac is adsorbed, and mixes with the feed liquid in the concentration tank again.When enzyme digestion reaction terminates, close ultrafiltration system, nanofiltration system continues operation, see through liquid and continue the blowback retort, meanwhile, in retort, add the soybean protein be used for next circulating reaction and with the suitable mother liquor of this circulation nanofiltration trapped fluid quantity, and begin to carry out and above-mentioned " the first step " identical operations.After treating the discharging of nanofiltration trapped fluid, carry out again and above-mentioned " second step " identical operations.
The molecular weight cut-off of nanofiltration membrane is chosen as 150～300; Activated carbon dosage is 0.5%～1.5% of the reaction water consumption; Nanofiltration pressure is chosen as 0.35mPa～0.50mPa; Time concentrated and the activated carbon decolorizing debitterize is generally 1h～5h; When nanofiltration concentrated solution volume is reduced to 5%～15% time discharging of water consumption, for carrying out drying.Can make the nanofiltration membrane flux reach coupling with higher level's ultra-filtration membrane flux by regulating pressure, to keep the synchronous operation of enzyme membrane reaction and nanofiltration separation.
The drying of the 3rd step soybean peptides
The nanofiltration trapped fluid that gives off is carried out conventional vacuum-drying or spray drying treatment to obtain the soybean peptides product of dry powdery.
It is 300～5000 soybean peptides that embodiment one soybean protein alkalescence enzymolysis prepares molecular weight
The first step is based on the enzyme membrane reaction of ultra-filtration and separation
The 500g soybean protein is dissolved in the distilled water of 5L and is mixed with 10% (W/V) soy bean proteinous soln, select the Alcalase Sumizyme MP for use, enzyme concentration 1.0 * 10
-2AU/g protein, mechanical stirring, rotating speed 50r/min, and regulate mother liquor pH=10 with the NaOH solution of 0.5mol/L makes the material system temperature reach 85 ℃ by interchanger, stirs to open ultrafiltration system behind the 1h and carry out enzyme membrane reaction, enzyme membrane reaction times 4h; Select that molecular weight cut-off is 5000, useful area is 0.7M for use
2The rolling ultra-filtration membrane, ultrafiltration pressure 0.08mPa; Beginning to carry out nanofiltration when concentration ratio reaches 1.5 handles.The trapped fluid of ultra-filtration process is sent back to and is continued to serve as reaction substrate in the retort, is used to carry out the concentrated and charcoal absorption processing of nanofiltration and send into concentration tank through liquid.
Charcoal absorption and the product of second step based on nanofiltration separation concentrates
Select that molecular weight cut-off is 300, useful area is 1.0M for use
2The rolling nanofiltration membrane, hanging basket is made with 100 eye mesh screens, charge weitght is 75g (reaction water consumption a 1.5%) oil charcoal in the hanging basket, the oil charcoal is selected particle between the sieve of 30 and 36 mesh sieves for use; Nanofiltration pressure 0.35mPa; The liquid pump that sees through of nanofiltration is got back to retort and is reused, and trapped fluid is flowed through after the hanging basket that is filled with the oil gac carries out charcoal absorption, mixes with the feed liquid in the concentration tank again; When enzyme digestion reaction termination (4h), close ultrafiltration system, nanofiltration system and charcoal absorption continue running, see through liquid and continue the blowback retort; Meanwhile, interpolation is used for the 500g soybean protein and the 500mL alkaline mother liquor of next circulating reaction in retort, and carries out and " the first step " identical operations; When nanofiltration concentrated solution volume was reduced to 10% (500mL) of water consumption, " second step " operation was carried out in discharging again.
The drying of the 3rd step soybean peptides
The nanofiltration trapped fluid (concentrated solution) that gives off through nanofiltration is carried out vacuum-drying, get the soybean peptides product of 62.3g dry powdery.
It is 300～3000 soybean peptides that the neutral enzymolysis of embodiment two soybean proteins prepares molecular weight
The first step is based on the enzyme membrane reaction of ultra-filtration and separation
The 300g soybean protein is dissolved in the deionized water of 10L and is mixed with 3% (W/V) soy bean proteinous soln, select the Neutrase neutral protease for use, enzyme concentration 3.0 * 10
-2AU/g protein, mechanical stirring, rotating speed 10r/min makes the material system temperature reach 50 ℃ by interchanger, opens ultrafiltration system behind the stirring 1h and carries out enzyme membrane reaction, enzyme membrane reaction times 6h; Select that molecular weight cut-off is 3000, useful area is 0.7M for use
2The rolling ultra-filtration membrane, ultrafiltration pressure 0.20mPa; Beginning to carry out nanofiltration when concentration ratio reaches 1.1 handles.The trapped fluid of ultra-filtration process is sent back to and is continued to serve as reaction substrate in the retort, is used to carry out the concentrated and charcoal absorption processing of nanofiltration and send into concentration tank through liquid.
Charcoal absorption and the product of second step based on nanofiltration separation concentrates
Select that molecular weight cut-off is 300, useful area is 1.0M for use
2The rolling nanofiltration membrane, hanging basket is made with 200 eye mesh screens, filling 50g in the hanging basket (reaction water consumption 0.5%) oil charcoal, oil charcoal are selected particle between the sieve of 24 and 30 mesh sieves for use; Nanofiltration pressure 0.40mPa; The liquid pump that sees through of nanofiltration is got back to retort and is reused, and trapped fluid is flowed through after the hanging basket that is filled with the oil charcoal carries out charcoal absorption, mixes with the feed liquid in the concentration tank again; When enzyme digestion reaction termination (6h), close ultrafiltration system, nanofiltration system and charcoal absorption continue running, see through liquid and continue the blowback retort; Meanwhile, in retort, add 300g soybean protein and the 500mL deionized water that is used for next circulating reaction, adjust pH=7.0, and carry out and " the first step " identical operations; When nanofiltration concentrated solution volume was reduced to 5% (500mL) of water consumption, " second step " operation was carried out in discharging again.
The drying of the 3rd step soybean peptides
The nanofiltration trapped fluid (concentrated solution) that gives off through nanofiltration is carried out spraying drying, get 53.5g powdered soybean peptides product.
It is 300～3000 soybean peptides that the acid enzymolysis of embodiment three soybean proteins prepares molecular weight
The first step is based on the enzyme membrane reaction of ultra-filtration and separation
The 600g soybean protein is dissolved in the distilled water of 10L and is mixed with 6% (W/V) soy bean proteinous soln, select the As3.350 aspartic protease for use, enzyme concentration 2.0 * 10
-2AU/g protein, mechanical stirring, rotating speed 100r/min, and regulate mother liquor pH=3.0 with the HCl solution of 1.0mol/L makes the material system temperature reach 55 ℃ by interchanger, stirs to open ultrafiltration system behind the 3h and carry out enzyme membrane reaction, enzyme membrane reaction times 8h; Selecting molecular weight cut-off for use is 3000, and useful area is 0.5M
2The cannulated ultra-filtration membrane, ultrafiltration pressure 0.12mPa; Beginning to carry out nanofiltration when concentration ratio reaches 2.0 handles.The trapped fluid of ultra-filtration process is sent back to and is continued to serve as reaction substrate in the retort, is used to carry out the concentrated and charcoal absorption processing of nanofiltration and send into concentration tank through liquid.
Charcoal absorption and the product of second step based on nanofiltration separation concentrates
Select that molecular weight cut-off is 300, useful area is 0.7M for use
2The cannulated nanofiltration membrane, hanging basket is made with 300 eye mesh screens, filling 100g in the hanging basket (reaction water consumption 1.0%) fiber-reactive charcoal, fiber-reactive charcoal are selected particle between the sieve of 50 and 100 mesh sieves for use; Nanofiltration pressure 0.50mPa; The liquid pump that sees through of nanofiltration is got back to retort and is reused, and trapped fluid is flowed through after the hanging basket that is filled with the fiber-reactive charcoal carries out charcoal absorption, mixes with the feed liquid in the concentration tank again; When enzyme digestion reaction termination (8h), close ultrafiltration system, nanofiltration system and charcoal absorption continue running, see through liquid and continue the blowback retort, meanwhile, in retort, add 600g soybean protein and the 1500mL distilled water that is used for next circulating reaction, adjust pH=3.0, and carry out and " the first step " identical operations; When nanofiltration concentrated solution volume was reduced to 15% (1500mL) of water consumption, " second step " operation was carried out in discharging again.
The drying of the 3rd step soybean peptides
The nanofiltration trapped fluid (concentrated solution) that gives off through nanofiltration is carried out spraying drying, get 82.4g powdered soybean peptides product.
Claims (5)
1. the method for the continuous enzyme membrane prepared in reaction of a soybean protein soybean peptides, with the soybean protein is raw material, react the soybean peptides solution that makes less than the ultra-filtration membrane molecular weight cut-off through enzyme membrane, after concentrate, dry powdery soybean peptide, it is characterized in that: soybean protein is dissolved in deionized water or the distilled water, quantity with soybean protein is that benchmark adds zymin, mechanical stirring, be mixed into reaction raw materials liquid, and, carry out enzyme membrane reaction based on ultra-filtration and separation with the pH value of acid solution or alkali lye conditioned reaction system; The trapped fluid of ultra-filtration process is sent back to and is continued to serve as reaction substrate in the retort, is used to carry out the concentrated and charcoal absorption processing of nanofiltration and see through liquid.
2. the method for the continuous enzyme membrane prepared in reaction of soybean protein as claimed in claim 1 soybean peptides, it is characterized in that in the reaction of soybean protein enzyme membrane, the weightmeasurement ratio of soybean protein addition and deionized water or distilled water is 3%～10%, the zymin addition with enzyme activity to soybean protein quality 1.0 * 10
-2AU/g～3.0 * 10
-2AU/g is a standard, pH3.0～10.0,50 ℃～85 ℃ of temperature of reaction; Stir back unlatching ultrafiltration system and carry out enzyme membrane reaction, the molecular weight cut-off 3000～5000 of ultra-filtration membrane, membrane filtration pressure 0.08mPa～0.30mPa, reaction times 4h～8h; Beginning to carry out nanofiltration when concentration ratio reaches 1.1～2.0 handles.
3. the method for the continuous enzyme membrane prepared in reaction of soybean protein as claimed in claim 2 soybean peptides is characterized in that zymin uses a kind of in Sumizyme MP, neutral protease or the aspartic protease.
4. the method for the continuous enzyme membrane prepared in reaction of soybean protein as claimed in claim 2 soybean peptides, it is characterized in that stirring used rotating speed is 10r/min～100r/min.
5. the method for the continuous enzyme membrane prepared in reaction of soybean protein as claimed in claim 1 or 2 soybean peptides, when it is characterized in that in nanofiltration separation concentrating with charcoal absorption and soybean peptides, the molecular weight cut-off of nanofiltration membrane is 150～300; The loadings of gac is 0.5%～1.5% of the reaction water consumption; Nanofiltration pressure 0.35mPa～0.50mPa; The liquid pump that sees through of nanofiltration is got back to the retort repeated use, and trapped fluid carries out mixing with the feed liquid in the concentration tank after the charcoal absorption again.
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| CNB2004100129663A CN100338224C (en) | 2004-04-01 | 2004-04-01 | Method for preparing peptide of soybean through continuous enzyme membrane reaction of soyabean protein |
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| CNB2004100129663A CN100338224C (en) | 2004-04-01 | 2004-04-01 | Method for preparing peptide of soybean through continuous enzyme membrane reaction of soyabean protein |
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| CN100338224C true CN100338224C (en) | 2007-09-19 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105506045A (en) * | 2016-01-12 | 2016-04-20 | 华隆（乳山）食品工业有限公司 | Efficient preparation technology for low-bitterness and high-quality soybean peptide |
| CN105950275A (en) * | 2016-05-17 | 2016-09-21 | 东北农业大学 | Aqueous enzymatic method for preparing blend sesame oil and composite protein peptide |
| CN106591116A (en) * | 2016-11-22 | 2017-04-26 | 大连大学 | Reaction device capable of eliminating inhibiting effect of enzymolysis product on enzymatic activity |
| CN108208309A (en) * | 2017-12-28 | 2018-06-29 | 保龄宝生物股份有限公司 | The production method of low ash content soybean peptide containing profitable probliotics |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07264993A (en) * | 1994-03-31 | 1995-10-17 | Fuji Oil Co Ltd | Production of soybean peptide mixture |
| CN1273783A (en) * | 2000-05-26 | 2000-11-22 | 黑龙江省轻工科学研究院 | Process for preparing active peptide of soybean protein |
| CN1403583A (en) * | 2002-07-12 | 2003-03-19 | 成都心友生物技术有限责任公司 | Prepn of natural active protein and peptide |
| CN1428431A (en) * | 2001-12-25 | 2003-07-09 | 山东力源集团公司 | Method for preparing soybean polypeptide by utilizing ereyme method |
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2004
- 2004-04-01 CN CNB2004100129663A patent/CN100338224C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07264993A (en) * | 1994-03-31 | 1995-10-17 | Fuji Oil Co Ltd | Production of soybean peptide mixture |
| CN1273783A (en) * | 2000-05-26 | 2000-11-22 | 黑龙江省轻工科学研究院 | Process for preparing active peptide of soybean protein |
| CN1428431A (en) * | 2001-12-25 | 2003-07-09 | 山东力源集团公司 | Method for preparing soybean polypeptide by utilizing ereyme method |
| CN1403583A (en) * | 2002-07-12 | 2003-03-19 | 成都心友生物技术有限责任公司 | Prepn of natural active protein and peptide |
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