CN103540528A - Enzyme-membrane coupling reaction system with speed-changing circulation - Google Patents
Enzyme-membrane coupling reaction system with speed-changing circulation Download PDFInfo
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- CN103540528A CN103540528A CN201310484423.0A CN201310484423A CN103540528A CN 103540528 A CN103540528 A CN 103540528A CN 201310484423 A CN201310484423 A CN 201310484423A CN 103540528 A CN103540528 A CN 103540528A
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- reaction
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
Abstract
The invention provides an enzyme-membrane coupling reaction system with speed-changing circulation and relates to the field of food processing and chemical engineering. The enzyme-membrane coupling reaction system is operated by the following steps of: putting a substrate solution with a certain concentration into a reaction kettle, adjusting the pH and the temperature of reaction liquid to set values, starting an ultrafiltration-device circulating pump to circulate for 5 minutes, stopping circulation, supplying feed liquid into the reactor to the initial liquid level, adding enzyme into a reaction kettle, starting the circulating pump again, performing enzymolysis-membrane separation coupling reaction, and collecting permeating liquid; for the enzyme-membrane coupling reaction system without supplying materials, after the reaction lasts for a period of time, reducing the rotating speed of the circulating pump to be 0.5-0.8 times of high speed till no products in the permeated liquid are discharged and ending the reaction; for the enzyme-membrane coupling reaction system with the continuous material supplying, when the products are discharged, continuously supplying the substrate solution into the reaction kettle at the same speed simultaneously, supplying clean water into the reaction kettle when ultrafiltration pressure rises to the limited value of a device, reducing the rotating speed of the circulating pump to be 0.5-0.8 times of the high speed after a period of time till no products in the permeating liquid are discharged, and ending the reaction.
Description
Technical field
The present invention relates to food processing field, refer in particular to a kind of method that strategy circulating by speed change improves enzyme membrane coupling reaction system substrate conversion efficiency of setting up.
Background technology
The main deficiency that traditional enzyme digestion reaction exists is: (1) Substrate inhibition: if the product of reaction is not removed as soon as possible, because product saturated reaction is suppressed, cause speed of response to decline; (2) utilising efficiency of enzyme is low: after a still reaction finishes, by the high temperature enzyme stopped reaction that goes out, start the reaction of next still, so the utilization ratio of enzyme is very low, causes production cost to strengthen; (3) the excessive enzymolysis of product: reached the enzymolysis product of optimum weight, owing to there is no to discharge and occur again digested situation in time, caused excessive enzyme to be cut and cause that activity declines; (4) substrate is not enough: arrived the reaction later stage, declining appears in amount of substrate, and the service efficiency of enzyme also will decline thereupon.
In order to solve traditional enzyme digestion reaction above shortcomings, many scholars highland barley seedlings by enzymolysis-membrane separation coupling reaction technology (being called for short " enzyme membrane coupling technique ") that begun one's study in recent years, by recycle pump, reactor and separatory membrane are coupled together, in enzyme digestion reaction, method by membrane sepn is isolated reactive system in time by the zymolyte that reaches molecular weight, simultaneously according to certain speed postreaction substrate continuously, until seriously stop up separatory membrane at the remaining reaction residues of reactive system, last termination reaction.From the description of above-mentioned working process, it seems, the advantage of enzyme membrane coupling technique maximum is to be exactly the index that represents enzyme utilization ratio---unit enzyme produces target compound amount and increases substantially.
But enzyme membrane coupling technique is still in some shortcomings at present, its working process and operating parameters still have the potentiality of further optimization.For example, the patent of invention that the inventor once applied for authorizing " gradient dilution feed supplement enzyme membrane coupling reaction and for scale collagen polypeptide the preparation " (patent No.: ZL201010253292.1) by the mode continuous feeding of gradient dilution, can significantly improve the working time of prolongation system, the unit's of increasing considerably enzyme produces target compound amount.
Summary of the invention
The object of the invention is to by further optimization enzyme membrane coupling reaction technology, improve the new technology of enzyme membrane coupling reaction system substrate conversion efficiency, be specifically just to provide a kind of first speed change cyclic policy of at a high speed rear low speed.
Its principle is: reaction utilizes Rapid Circulation early stage, and enzymolysis small molecules product is out discharged to reactive system in time eliminates rapidly product retarding effect on the one hand, and reinforcing mass transfer effect promotes the abundant contact of enzyme-to-substrate on the other hand, thereby improves enzymolysis efficiency; In the reaction later stage, it is not enough that substrate becomes, and circulation fast becomes on the contrary and reduces effective contact of enzyme-to-substrate, therefore reduces the carrying out that rotating speed is conducive to enzyme digestion reaction, and the transformation efficiency of substrate still can continue to keep a higher level.
The invention provides a kind of speed change circulation enzyme membrane coupling reaction system, according to following step work:
With the enzymolysis process of scale collagen polypeptide, being prepared as example is described.
(1) for the enzyme membrane coupling reaction system of no-feed supplement
1) in reactor, add substrate solution;
2) enzyme-addedly carry out the reaction of highland barley seedlings by enzymolysis-membrane separation coupling;
3) reduce recycle pump rotating speed, until reaction finishes.
In step 1 of the present invention) in, pack the fish scale collagen solution of certain concentration of substrate into reactor, regulate reaction solution pH and temperature to the value of setting, open ultra-filtration equipment recycle pump circulation 5min, stop circulation device, now ultra-filtration equipment and pipeline are all full of feed liquid.
In step 2 of the present invention) in, in reactor, supplement feed liquid to the liquid level starting again, and according to the ratio of enzyme-to-substrate, add appropriate proteolytic enzyme in reactor, ON cycle pump again, start the reaction of highland barley seedlings by enzymolysis-membrane separation coupling, in reaction process, keep enzyme digestion reaction temperature and reacting liquid pH value, collect and see through liquid.
In step 3 of the present invention) in, after reaction for some time, recycle pump rotating speed is reduced to at a high speed 0.5～0.8 times, until see through in liquid, without polypeptide, discharge, stop this reaction.
(2) for the enzyme membrane coupling reaction system of continuous feeding
1) in reactor, add substrate solution;
2) enzyme-addedly carry out the reaction of highland barley seedlings by enzymolysis-membrane separation coupling, and continuous supplementation substrate solution;
3) later stage moisturizing, reduces recycle pump rotating speed, until reaction finishes.
In step 1 of the present invention) in, pack the fish scale collagen solution of certain concentration of substrate into reactor, regulate reaction solution pH and temperature to the value of setting, open ultra-filtration equipment recycle pump circulation 5min, stop circulation device, now ultra-filtration equipment and pipeline are all full of feed liquid.
In step 2 of the present invention) in, in reactor, supplement feed liquid to the liquid level starting again, and according to the ratio of enzyme-to-substrate, add appropriate proteolytic enzyme in reactor, ON cycle pump again, start the reaction of highland barley seedlings by enzymolysis-membrane separation coupling, in reaction process, keep enzyme digestion reaction temperature and reacting liquid pH value, collect and see through liquid.In row's peptide, with same speed to continuous supplementation fish scale collagen solution in reactor, to keep reactor liquid level constant.
In step 3 of the present invention) in, ultrafiltration pressure rises to the limit value of device, starts to supplement clear water in reactor, after for some time, recycle pump rotating speed is reduced to at a high speed 0.5～0.8 times, until see through in liquid, without polypeptide, discharges, and stops this reaction.
Advantage of the present invention is: by the later stage, reduce the mode of speed of circulation, improve the transformation efficiency of enzyme membrane coupling reaction substrate, improve reaction efficiency.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but working of an invention mode is not limited to this.
Embodiment 1
Getting concentration and be 3% fish scale collagen solution 400mL adds in enzymolysis reactor, regulate reaction solution pH9.0, regulate 55 ℃ of temperature of reaction, open ultra-filtration equipment recycle pump, allow the Mierocrystalline cellulose flat sheet membrane that protein solution is 3kDa by molecular weight cut-off, after 5min, stop circulation, now ultra-filtration equipment and pipeline are all full of feed liquid.Again to supplementing feed liquid in reactor to the liquid level starting, and to the protease A lcalase2.4L that adds substrate 1.5% in reactor.ON cycle pump again, feed liquid, with the rotating speed circulation of 150r/min, starts the reaction of highland barley seedlings by enzymolysis-membrane separation coupling, keeps enzyme digestion reaction temperature and reacting liquid pH value in reaction process, collects and sees through liquid.After reaction 15min, recycle pump rotating speed is reduced to 98r/min, continues coupling reaction 20min, termination reaction.Measure and find, it is 143g that the transformation efficiency 96%， unit enzyme of protein produces peptide amount.
Embodiment 2
Getting concentration and be 3% fish scale collagen solution 400mL adds in enzymolysis reactor, regulate reaction solution pH9.0, regulate 55 ℃ of temperature of reaction, open ultra-filtration equipment recycle pump, allow the Mierocrystalline cellulose flat sheet membrane that protein solution is 3kDa by molecular weight cut-off, after 5min, stop circulation, now ultra-filtration equipment and pipeline are all full of feed liquid.Again to supplementing feed liquid in reactor to the liquid level starting, and to the protease A lcalase2.4L that adds substrate 1.5% in reactor.ON cycle pump again, feed liquid, with the rotating speed circulation of 170r/min, starts the reaction of highland barley seedlings by enzymolysis-membrane separation coupling, keeps enzyme digestion reaction temperature and reacting liquid pH value in reaction process, collects and sees through liquid.After reaction 15min, recycle pump rotating speed is reduced to 85r/min, continues coupling reaction 25min, termination reaction.Measure and find, it is 139g that the transformation efficiency 91%， unit enzyme of protein produces peptide amount.
Embodiment 3
Getting concentration and be 3% fish scale collagen solution 400mL adds in enzymolysis reactor, regulate reaction solution pH9.0, regulate 55 ℃ of temperature of reaction, open ultra-filtration equipment recycle pump, allow the Mierocrystalline cellulose flat sheet membrane that protein solution is 3kDa by molecular weight cut-off, after 5min, stop circulation, now ultra-filtration equipment and pipeline are all full of feed liquid.Again to supplementing feed liquid in reactor to the liquid level starting, and to the protease A lcalase2.4L that adds substrate 1.5% in reactor.ON cycle pump again, feed liquid, with the rotating speed circulation of 150r/min, starts the reaction of highland barley seedlings by enzymolysis-membrane separation coupling, keeps enzyme digestion reaction temperature and reacting liquid pH value in reaction process, collects and sees through liquid.In row's peptide, with same speed to continuous supplementation fish scale collagen solution in reactor, to keep reactor liquid level constant.When ultrafiltration pressure rises to the limit value of device, start to supplement clear water in reactor, after 10min, recycle pump rotating speed is reduced to 100%, until see through in liquid, without polypeptide, discharge, stop this reaction.Measure and find, the transformation efficiency 95%， unit enzyme of protein produces peptide amount 258g.
Embodiment 4
Getting concentration and be 3% fish scale collagen solution 400mL adds in enzymolysis reactor, regulate reaction solution pH9.0, regulate 55 ℃ of temperature of reaction, open ultra-filtration equipment recycle pump, allow the Mierocrystalline cellulose flat sheet membrane that protein solution is 3kDa by molecular weight cut-off, after 5min, stop circulation, now ultra-filtration equipment and pipeline are all full of feed liquid.Again to supplementing feed liquid in reactor to the liquid level starting, and to the protease A lcalase2.4L that adds substrate 1.5% in reactor.ON cycle pump again, feed liquid, with the rotating speed circulation of 150r/min, starts the reaction of highland barley seedlings by enzymolysis-membrane separation coupling, keeps enzyme digestion reaction temperature and reacting liquid pH value in reaction process, collects and sees through liquid.In row's peptide, with same speed to continuous supplementation fish scale collagen solution in reactor, to keep reactor liquid level constant.When ultrafiltration pressure rises to the limit value of device, start to supplement clear water in reactor, after 15min, recycle pump rotating speed is reduced to 120%, until see through in liquid, without polypeptide, discharge, stop this reaction.Measure and find, the transformation efficiency 98%， unit enzyme of protein produces peptide amount 263g.
Claims (2)
1. a speed change circulation enzyme membrane coupling reaction system, is characterized in that reaction solution speed change circulation between reactor and separatory membrane in highland barley seedlings by enzymolysis-membrane separation coupling reactive system.
2. a kind of speed change circulation enzyme membrane coupling reaction system according to claim 1, is characterized in that reaction solution takes the shift strategy of low speed circulation after first high-speed circulating, and low speed is at a high speed 0.5～0.8 times.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109097408A (en) * | 2018-09-03 | 2018-12-28 | 河北美邦工程科技股份有限公司 | A kind of preparation method of 56 salt of nylon |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101284017A (en) * | 2008-05-27 | 2008-10-15 | 浙江大学 | A method for continuously preparing birds nest extract with narrow molecular weight distribution by enzymolysis and membrane filtration coupling technique |
| CN101591614A (en) * | 2009-06-23 | 2009-12-02 | 江苏大学 | A kind of ultrasonic enzymatic membrane reactor |
| CN101845472A (en) * | 2010-05-28 | 2010-09-29 | 江苏大学 | Method for preparing scale collagen antioxidant peptides by using no-feed supplement enzymolysis-membrane separation coupling technology |
| CN101948894A (en) * | 2010-08-13 | 2011-01-19 | 江苏大学 | Gradient dilution feed enzyme membrane coupling reaction and application thereof for preparing scale collagen polypeptide |
| CN102212599A (en) * | 2011-04-29 | 2011-10-12 | 北京工商大学 | Method for preparing oat ACE (Angiotensin Converting Enzyme) inhibitory peptide by using enzymatic membrane reactor |
-
2013
- 2013-10-17 CN CN201310484423.0A patent/CN103540528A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101284017A (en) * | 2008-05-27 | 2008-10-15 | 浙江大学 | A method for continuously preparing birds nest extract with narrow molecular weight distribution by enzymolysis and membrane filtration coupling technique |
| CN101591614A (en) * | 2009-06-23 | 2009-12-02 | 江苏大学 | A kind of ultrasonic enzymatic membrane reactor |
| CN101845472A (en) * | 2010-05-28 | 2010-09-29 | 江苏大学 | Method for preparing scale collagen antioxidant peptides by using no-feed supplement enzymolysis-membrane separation coupling technology |
| CN101948894A (en) * | 2010-08-13 | 2011-01-19 | 江苏大学 | Gradient dilution feed enzyme membrane coupling reaction and application thereof for preparing scale collagen polypeptide |
| CN102212599A (en) * | 2011-04-29 | 2011-10-12 | 北京工商大学 | Method for preparing oat ACE (Angiotensin Converting Enzyme) inhibitory peptide by using enzymatic membrane reactor |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109097408A (en) * | 2018-09-03 | 2018-12-28 | 河北美邦工程科技股份有限公司 | A kind of preparation method of 56 salt of nylon |
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Application publication date: 20140129 |