CN101768551A - Multipole immobilized enzyme film reaction device and method of preparing casein phosphopeptides by using reaction device - Google Patents
Multipole immobilized enzyme film reaction device and method of preparing casein phosphopeptides by using reaction device Download PDFInfo
- Publication number
- CN101768551A CN101768551A CN201019026024.3A CN201019026024A CN101768551A CN 101768551 A CN101768551 A CN 101768551A CN 201019026024 A CN201019026024 A CN 201019026024A CN 101768551 A CN101768551 A CN 101768551A
- Authority
- CN
- China
- Prior art keywords
- grade
- fixed enzyme
- enzyme membrane
- membrane reactor
- molecular weight
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108010001441 Phosphopeptides Proteins 0.000 title claims abstract description 54
- 238000006243 chemical reaction Methods 0.000 title claims abstract description 49
- 239000005018 casein Substances 0.000 title claims abstract description 26
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 title claims abstract description 26
- 235000021240 caseins Nutrition 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 19
- 108010093096 Immobilized Enzymes Proteins 0.000 title abstract description 5
- 230000005405 multipole Effects 0.000 title abstract 2
- 239000012528 membrane Substances 0.000 claims abstract description 120
- 238000001728 nano-filtration Methods 0.000 claims abstract description 50
- 230000007423 decrease Effects 0.000 claims abstract description 5
- 102000004190 Enzymes Human genes 0.000 claims description 118
- 108090000790 Enzymes Proteins 0.000 claims description 118
- 229940088598 enzyme Drugs 0.000 claims description 116
- 229940071162 caseinate Drugs 0.000 claims description 47
- 238000000108 ultra-filtration Methods 0.000 claims description 23
- 239000007788 liquid Substances 0.000 claims description 21
- 108091005804 Peptidases Proteins 0.000 claims description 12
- 239000000463 material Substances 0.000 claims description 11
- 238000003756 stirring Methods 0.000 claims description 11
- 102000035195 Peptidases Human genes 0.000 claims description 10
- 238000010612 desalination reaction Methods 0.000 claims description 9
- 238000001976 enzyme digestion Methods 0.000 claims description 8
- 239000002131 composite material Substances 0.000 claims description 5
- 239000004952 Polyamide Substances 0.000 claims description 3
- 239000004365 Protease Substances 0.000 claims description 3
- 239000004745 nonwoven fabric Substances 0.000 claims description 3
- 229920002492 poly(sulfone) Polymers 0.000 claims description 3
- 229920002647 polyamide Polymers 0.000 claims description 3
- 229920000728 polyester Polymers 0.000 claims description 3
- 239000011148 porous material Substances 0.000 claims description 3
- 108090000145 Bacillolysin Proteins 0.000 claims description 2
- 102000035092 Neutral proteases Human genes 0.000 claims description 2
- 108091005507 Neutral proteases Proteins 0.000 claims description 2
- 108010019160 Pancreatin Proteins 0.000 claims description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 2
- 229920002301 cellulose acetate Polymers 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims description 2
- 229940055695 pancreatin Drugs 0.000 claims description 2
- 229920000515 polycarbonate Polymers 0.000 claims description 2
- 239000004417 polycarbonate Substances 0.000 claims description 2
- 230000035484 reaction time Effects 0.000 claims description 2
- -1 papoid Proteins 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 12
- 239000000243 solution Substances 0.000 description 23
- 108010076119 Caseins Proteins 0.000 description 20
- 238000006460 hydrolysis reaction Methods 0.000 description 19
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 18
- 230000007062 hydrolysis Effects 0.000 description 15
- 239000011575 calcium Substances 0.000 description 9
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 8
- 229910052791 calcium Inorganic materials 0.000 description 8
- 239000000758 substrate Substances 0.000 description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 230000008021 deposition Effects 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 238000001879 gelation Methods 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 210000000813 small intestine Anatomy 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 239000011701 zinc Substances 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 1
- 206010065687 Bone loss Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- BZQFBWGGLXLEPQ-UHFFFAOYSA-N O-phosphoryl-L-serine Natural products OC(=O)C(N)COP(O)(O)=O BZQFBWGGLXLEPQ-UHFFFAOYSA-N 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 102000005158 Subtilisins Human genes 0.000 description 1
- 108010056079 Subtilisins Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 208000012826 adjustment disease Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 108010079058 casein hydrolysate Proteins 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 208000002925 dental caries Diseases 0.000 description 1
- 229950006137 dexfosfoserine Drugs 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 230000008175 fetal development Effects 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910001463 metal phosphate Inorganic materials 0.000 description 1
- 108010009355 microbial metalloproteinases Proteins 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- BZQFBWGGLXLEPQ-REOHCLBHSA-N phosphoserine Chemical compound OC(=O)[C@@H](N)COP(O)(O)=O BZQFBWGGLXLEPQ-REOHCLBHSA-N 0.000 description 1
- 239000012716 precipitator Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 208000007442 rickets Diseases 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Classifications
-
- 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
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/18—Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
-
- 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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
-
- 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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/58—Reaction vessels connected in series or in parallel
-
- 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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/02—Membranes; Filters
-
- 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
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
- C12M41/22—Heat exchange systems, e.g. heat jackets or outer envelopes in contact with the bioreactor walls
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Clinical Laboratory Science (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Thermal Sciences (AREA)
- Analytical Chemistry (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention discloses a multipole immobilized enzyme film reaction device, comprising an immobilized enzyme film reactor connected in series of N stages and a nanofiltration device; a cylinder is installed in each stage of immobilized enzyme film reactors; film with cut-off molecular weight of 1000- 10000Da is filled in inside of the cylinder; prolease is fixed on the surface of inside of the film; nanofiltration membrane with cut-off molecular weight of 100- 500Da is installed in the nanofiltration device; the outside part of the first stage reactor cylinder is connected with the inside part of the second stage reactor cylinder, by parity of reasoning, the outside part of the N stage reactor cylinder is connected with the nanofiltration device, the inside parts of the second stage to the N stage reactor cylinders are respectively connected with the nanofiltration device; the cut-off molecular weights of film decrease gradually from the first stage to the N stage reactor. The present invention also discloses a method of preparing casein phosphopeptides by using the reaction device. Casein phosphopeptides with different molecular weight and quality can be prepared by the present invention, course of production is simplified, and cost of production is reduced.
Description
Technical field
The present invention relates to the preparation facilities and the method for phosphopeptide caseinate, be specifically related to a kind of multistage fixed enzyme membrane reaction unit and utilize it to prepare the method for phosphopeptide caseinate.
Background technology
Phosphopeptide caseinate (casein phosphopeptide is CPPs) is to be a kind of natural bioactive polypeptide that contains the cluster phosphoserine that raw material makes through single or prozyme catalytic hydrolysis, separation, purifying with the bovine casein.
Phosphopeptide caseinate can be used as the carrier of mineral ion, and it is neutral to subalkaline animal small intestine lower end at pH, energy and Ca
2+, Fe
2+And Zn
2+Deng the metal ion combination, stop metal ion and phosphate radical generation precipitation, calcium, iron and the zinc concentration of solubility in the small intestine are increased, thereby promote these essential metal ionic to absorb and utilization.In addition, in animal and human's experiment, CPPs shows anti-dental caries, anti-bone loss, strengthens effects such as fertilization and fetal development.
China resident average every day of calcium intake has only 405.4mg, only account for about 50% of recommended amounts every day, up to 40%, osteoporosis also is one of common disease of puzzlement old man at the sickness rate of some regional children rachitis, therefore concerning China, the whole people replenish the calcium and seem and be even more important.And not only to pay attention to the absorption of calcium when replenishing the calcium, more to pay attention to the absorption of calcium.And CPPs be owing to can promote this function of calcium absorption, and its market application foreground is quite wide.
The core texture of phosphopeptide caseinate is-Ser (P)-Ser (P)-Ser (P)-Glu-Glu-, and this sequence is the functional gene of CPPs just also, and what of its content are directly related with the functional performance of CPPs.Therefore, the CPPs of different molecular weight shows different qualities and functional performance.At present, there is not a kind of method to control to the phosphopeptide caseinate molecular weight product.
At present, the main production method of phosphopeptide caseinate is an enzymolysis process, promptly in common hydrolytic decomposition pot, the substrate casein is hydrolyzed with proteolytic enzyme, after hydrolysis finishes, the high temperature enzyme that goes out, re-adjustment reaction solution pH makes unhydrolysed casein precipitation, and the centrifugal back spraying drying or the use calcium ethylate precipitator method make.Mainly there are the following problems in the technological process: (1) enzyme can not recycle, and the consumption of enzyme amount is big, the production cost height.(2) substrate hydrolysis is incomplete, and unreacted substrate is removed by centrifugal back, the waste raw materials for production.(3) product purity is low, and the product purity that obtains after the general spraying drying only about 12%, does not satisfy market demands fully.Have report to prepare phosphopeptide caseinate, but its proteolytic enzyme is not immobilized, and can not reuse, and has increased production cost with the Reaction Separation coupling process yet.(4) can not control product quality, can not control the molecular weight of phosphopeptide caseinate.
Summary of the invention
Technical problem to be solved by this invention provides a kind of multistage fixed enzyme membrane reaction unit, utilizes this reactor can prepare the product of different molecular weight simultaneously.
The technical problem that the present invention also will solve provides utilizes above-mentioned multistage fixed enzyme membrane reactor to prepare the method for phosphopeptide caseinate, this method can be prepared the phosphopeptide caseinate of different molecular weight and different quality purity simultaneously, has improved production efficiency greatly.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
A kind of multistage fixed enzyme membrane reaction unit, it comprises the placed in-line fixed enzyme membrane reactor of N level and a nanofiltration device; Each grade fixed enzyme membrane reactor forms cylinder with the polysulfones porous material for being supported on inside reactor, be divided into reactor in the cylinder and outer two spaces of cylinder, strengthen the intensity of above-mentioned cylindrical structure with polyester non-woven fabric, with molecular weight cut-off is the inboard that the film of 1000～10000Da is seated in above-mentioned cylindrical structure, is fixed with proteolytic enzyme on the film inner surface; Be provided with the nanofiltration membrane that molecular weight cut-off is 100～500Da in the nanofiltration device; The 1st grade of fixed enzyme membrane reactor cylindrical outer is connected with the 2nd grade of fixed enzyme membrane reactor cylinder interior by pipeline, the 2nd grade of fixed enzyme membrane reactor cylindrical outer is connected with 3rd level fixed enzyme membrane reactor cylinder interior by pipeline, by that analogy, until N level fixed enzyme membrane reactor, N level fixed enzyme membrane reactor cylindrical outer is connected with nanofiltration device by pipeline, the 2nd grade is connected with nanofiltration device by pipeline respectively to N level fixed enzyme membrane reactor cylinder interior, and above-mentioned pipeline all is provided with valve; The 1st grade of fixed enzyme membrane reactor to the N level fixed enzyme membrane reactor, the molecular weight cut-off of its film successively decreases successively; Described N gets 2～100 integer.
Wherein, described molecular weight cut-off is that the film of 1000～10000Da is any one in polyamide composite film, cellulose acetate film and the polycarbonate composite membrane.
Wherein, described fixed enzyme membrane reactor is equipped with the temperature control chuck.
Wherein, described fixed enzyme membrane reactor cylinder interior is provided with stirring rake.
Wherein, described proteolytic enzyme is any one or a few in Sumizyme MP (Alcalase), pancreatin (Trypsin), papoid (Papain), neutral protease (Neutrase), the compound protease (Protemax).
The method that above-mentioned proteolytic enzyme is fixed on the film is: adopt gelling technique that proteolytic enzyme is immobilized on the film inner surface.Be 0.01～0.08M Pa at working pressure promptly with membrane module, ultrafiltration protein enzyme solution under the condition of circular flow 10～30L/h, because the enzyme molecule can not be through film and at the film surface deposition, when the concentration of film-liquid two-phase interface place zymoprotein reaches gelation concentration, enzyme just is fixed on the surface of film with the form of a film gel coat, thereby realizes the immobilization of enzyme on film.Described protein enzyme solution is by making in the phosphoric acid buffer that proteolytic enzyme is dissolved in pH6.0～9.0.
Wherein, described N gets 2～5 integer.
Utilize above-mentioned multistage fixed enzyme membrane reaction unit to prepare the method for phosphopeptide caseinate, casein solution is injected into the 1st grade of fixed enzyme membrane reactor cylinder interior, carry out enzyme digestion reaction, after reaction finishes, by ultrafiltration molecular weight in the cylinder interior liquid is transformed into cylindrical outer less than the material of the 1st grade of membrane molecule amount, by pipeline the material of the 1st grade of fixed enzyme membrane reactor cylindrical outer is injected the 2nd grade of fixed enzyme membrane reactor cylinder interior again, carry out enzyme digestion reaction, after reaction finishes, by ultrafiltration molecular weight in the cylinder interior liquid is transformed into cylindrical outer less than the material of the 2nd grade of membrane molecule amount, by that analogy, until N level fixed enzyme membrane reactor, N level fixed enzyme membrane reactor cylindrical outer liquid is injected nanofiltration device by pipeline, obtain molecular weight less than N level membrane molecule amount and greater than the phosphopeptide caseinate of nanofiltration membrane molecular weight through the nanofiltration desalination and concentration; Simultaneously can be after the 2nd grade of each grade fixed enzyme membrane reactor to the N level carries out ultrafiltration the material of this grade fixed enzyme membrane reactor cylinder interior be injected nanofiltration device, obtain molecular weight less than one-level membrane molecule amount before this grade and greater than the phosphopeptide caseinate of this grade membrane molecule amount, the 1st grade of fixed enzyme membrane reactor cylinder interior can be replenished the casein solution reaction repeated through the nanofiltration desalination and concentration.
Wherein, described casein solution, concentration are 40～150g/L, and pH is 6.5～9.5.
Wherein, the temperature of reaction of each grade fixed enzyme membrane reactor is 30～60 ℃, and stirring velocity is 50～250rpm, and the enzyme digestion reaction time is 1～3h.
Wherein, each grade fixed enzyme membrane reactor ultrafiltration pressure is 0.01～0.08M Pa.
Above-mentioned reaction can be carried out continuously, also can intermittence carry out.
Beneficial effect: compared with prior art, advantage of the present invention is:
(1) adopts gelling technique that enzyme is fixed on and form fixed enzyme membrane reactor on the film, realize the recycling of enzyme, reduced production cost.
(2) immobilized enzyme mebrane reactor collection production separation and one, realization response catalysis synchronously separates with product, has simplified the production stage of phosphopeptide caseinate, has improved production efficiency.
(3) connect by the enzyme mebrane reactor of different progression, and with the ultra-filtration membrane of PSPP combination separates phosphopeptide caseinate with nanofiltration membrane, the phospho-peptide product of different molecular weight size can be obtained simultaneously, the product of different qualities can be produced at any time according to customer requirement.
(4) saved raw material, the substrate casein that is not hydrolyzed in the A reactor is utilized when reacting for the second time, has avoided the waste of raw material, has directly reduced reaction cost.
(5) reaction conditions gentleness involved in the present invention, energy consumption is low, and fixed enzyme membrane reactor can be reused and serialization production, improves the utilization ratio of enzyme and the purity of product greatly, is fit to the requirement of large-scale continuous production.
Description of drawings
Fig. 1 is a multistage fixed enzyme membrane reaction unit synoptic diagram of the present invention.
Wherein 1 is the temperature control chuck; 2 is stirring rake; 3 are membrane module (being cylinder); 4 is fixed enzyme membrane reactor; 5 is the product storage tank; 6 is valve; 7 nanofiltration device.
Embodiment
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand that the described concrete material proportion of embodiment, processing condition and result thereof only are used to illustrate the present invention, and should also can not limit the present invention described in detail in claims.
Embodiment 1:
Fig. 1 is a specific embodiment of the present invention, and as can be seen from the figure, this multistage fixed enzyme membrane reaction unit comprises 3 grades of placed in-line fixed enzyme membrane reactors 4 and a nanofiltration device; Each grade fixed enzyme membrane reactor forms cylinder 3 with the polysulfones porous material for being supported on inside reactor, be divided into reactor in the cylinder and outer two spaces of cylinder, strengthen the intensity of above-mentioned cylindrical structure with polyester non-woven fabric, with molecular weight cut-off is the inboard that the film of 1000～10000Da is seated in above-mentioned cylindrical structure, is fixed with proteolytic enzyme on the film inner surface; Be provided with the nanofiltration membrane that molecular weight cut-off is 100～500Da in the nanofiltration device; The 1st grade of fixed enzyme membrane reactor cylindrical outer is connected with the 2nd grade of fixed enzyme membrane reactor cylinder interior by pipeline, and by the folding of valve 6a control pipeline; The 2nd grade of fixed enzyme membrane reactor cylindrical outer is connected with 3rd level fixed enzyme membrane reactor cylinder interior by pipeline, and by the folding of valve 6b control pipeline; 3rd level fixed enzyme membrane reactor cylindrical outer is connected with nanofiltration device 7 by pipeline, and by the folding of valve 6c control pipeline; The 2nd grade of fixed enzyme membrane reactor cylinder interior is connected with nanofiltration device 7 by pipeline, and by the folding of valve 6d control pipeline; 3rd level fixed enzyme membrane reactor cylinder interior is connected with nanofiltration device 7 by pipeline, and by the folding of valve 6e control pipeline; The 1st grade of fixed enzyme membrane reactor is to the 3rd level fixed enzyme membrane reactor, and the molecular weight cut-off of its film successively decreases successively.Each fixed enzyme membrane reactor also is equipped with temperature control chuck 1, and is provided with stirring rake 2 in cylinder interior.
Fig. 1 has only shown one embodiment of the present invention, certainly the quantity of fixed enzyme membrane reactor is not only 3, can be any more than 2, no matter how the quantity of fixed enzyme membrane reactor changes, its annexation and catenation principle are similar to Fig. 1 device shown, and those skilled in the art are readily appreciated that, according to the structural principle of Fig. 1, N fixed enzyme membrane reactor and nanofiltration device are coupled together, thereby realize purpose of the present invention.
According to the device of Fig. 1, with molecular weight cut-off be 8000,6000, the polyamide composite film of 3000Da places the 1st, 2,3 grade of fixed enzyme membrane reactor cylinder inboard respectively, is that the nanofiltration membrane of 350Da places nanofiltration device with molecular weight cut-off.Certainly, the present invention is not limited to the film of above-mentioned molecular weight cut-off and above-mentioned material, and the film of any molecular weight cut-off and material can be applicable to the present invention, successively decreases successively by the liquid flow path direction as long as guarantee the molecular weight cut-off of film.
Take by weighing the 5g papoid, be dissolved in the phosphoric acid buffer of 1L pH 7.0, enzyme liquid is poured in the fixed enzyme membrane reactor cylinder, at working pressure 0.05M Pa, carry out ultrafiltration under the condition of circular flow 10L/h, because the enzyme molecule can not be through film and at the film surface deposition, when the concentration of film-liquid two-phase interface place enzyme molecule reaches gelation concentration, enzyme just is fixed on the surface of film with the form of a film gel coat, thereby realizes the immobilization of enzyme on film.Certainly the kind of enzyme also is not limited to above-mentioned papoid, only is that to have caseinhydrolysate be that the enzyme of phosphopeptide caseinate ability can use, and all in protection scope of the present invention.
Following examples are all used the device in the present embodiment.
Embodiment 2:
Take by weighing the 500g casein, add 10L 0.05M NaOH solution and place container for storing liquid, heating is also constantly stirred and is made casein dissolve fully, treats that casein dissolves the back fully and with 5M NaOH solution the pH of reaction solution transferred to 7.0.
Casein solution is injected in the 1st grade of fixed enzyme membrane reactor cylinder, the ON cycle cooling/heating apparatus, temperature of reaction is controlled at 50 ℃, open stirring rake simultaneously, the adjusting mixing speed is 100rpm, the beginning enzyme digestion reaction, the pH of reaction solution is constant in 7.0 in the reaction process with 5M NaOH solution.Behind the hydrolysis 2h, open ultra-filtration equipment, ultrafiltration pressure is 0.05Mpa, circular flow 10L/h.The phosphopeptide caseinate that the molecular weight that hydrolysis is produced is lower than 8000Da sees through, be transformed into the outside of cylinder, Open valve 6a, the phosphopeptide caseinate that makes molecular weight be lower than 8000Da flow in the 2nd grade of fixed enzyme membrane reactor cylinder, see through the substrate casein of the 1st grade of film and macromole polypeptide and then stay and continue hydrolysis in the enzyme mebrane reactor, the casein solution added with the pH 7.0 of amount of the amount of liquid that sees through according to ultrafiltration continues reaction simultaneously.
After treating that the interior liquid of the 2nd grade of fixed enzyme membrane reactor cylinder reaches about 5L, open the 2nd grade of stirring rake, the beginning hydrolysis reaction, reaction conditions is identical with the 1st grade.Hydrolysis was opened ultra-filtration equipment after 1 hour approximately, and ultrafiltration pressure is 0.05Mpa, circular flow 10L/h.The phosphopeptide caseinate that the molecular weight that hydrolysis is produced is lower than 6000Da sees through, and is transformed into the outside of cylinder, and Open valve 6b, the phosphopeptide caseinate that makes molecular weight be lower than 6000Da flow in the 3rd level fixed enzyme membrane reactor cylinder.In the 2nd grade of fixed enzyme membrane reactor cylinder is the phosphopeptide caseinate of molecular weight between 6000～8000Da.
After treating that the interior liquid of 3rd level fixed enzyme membrane reactor cylinder reaches about 3L, open the 3rd level stirring rake, the beginning hydrolysis reaction, reaction conditions is identical with the 1st grade.Hydrolysis was opened ultra-filtration equipment after 1 hour approximately, and ultrafiltration pressure is 0.05Mpa, circular flow 10L/h.The phosphopeptide caseinate that the molecular weight that hydrolysis is produced is lower than 3000Da sees through, and is transformed into the outside of cylinder, Open valve 6c, and the phosphopeptide caseinate that makes molecular weight be lower than 3000Da flow in the nanofiltration device.In the 2nd grade of fixed enzyme membrane reactor cylinder is the phosphopeptide caseinate of molecular weight between 3000～6000Da.
To open nanofiltration device, and be used for concentrating and desalination of product, working pressure 0.15Mpa when dehydration rate reaches 50%, finishes nanofiltration.The nanofiltration concentrated solution is the phosphopeptide caseinate product of molecular weight at 350～3000Da.
Also but Open valve 6d directly injects nanofiltration device with liquid in the 2nd grade of fixed enzyme membrane reactor cylinder, obtains the phosphopeptide caseinate product of molecular weight at 6000～8000Da through the nanofiltration desalination and concentration.
Also but Open valve 6e directly injects nanofiltration device with liquid in the 3rd level fixed enzyme membrane reactor cylinder, obtains the phosphopeptide caseinate product of molecular weight at 3000～6000Da through the nanofiltration desalination and concentration.
After testing, the index of above-mentioned 3 kinds of phosphopeptide caseinates is as follows:
The phosphopeptide caseinate of molecular weight 8000～6000Da: purity: 22.81%, protein content: 93.07%, N content: P content 14.59%: 0.99%;
The phosphopeptide caseinate of molecular weight 3000～6000Da: purity: 44.19%, protein content: 92.40%, N content: P content 14.48%: 1.47%;
The phosphopeptide caseinate of molecular weight 350～3000Da: purity: 67.25%, protein content: 91.71%, N content: P content 14.37%: 1.86%.
Embodiment 3:
Take by weighing the 400g casein, add 8L 0.05M NaOH solution and place container for storing liquid, heating is also constantly stirred and is made casein dissolve fully, treats that casein dissolves the back fully and with 5M NaOH solution the pH of reaction solution transferred to 7.0.
Casein solution is injected in the 1st grade of fixed enzyme membrane reactor cylinder, the ON cycle cooling/heating apparatus, temperature of reaction is controlled at 55 ℃, open stirring rake simultaneously, the adjusting mixing speed is 150rpm, the beginning enzyme digestion reaction, the pH of reaction solution is constant in 8.0 in the reaction process with 5M NaOH solution.Behind the hydrolysis 3h, open ultra-filtration equipment, ultrafiltration pressure is 0.05Mpa, circular flow 10L/h.The phosphopeptide caseinate that the molecular weight that hydrolysis is produced is lower than 8000Da sees through, be transformed into the outside of cylinder, Open valve 6a, the phosphopeptide caseinate that makes molecular weight be lower than 8000Da flow in the 2nd grade of fixed enzyme membrane reactor cylinder, see through the substrate casein of the 1st grade of film and macromole polypeptide and then stay and continue hydrolysis in the enzyme mebrane reactor, the casein solution added with the pH 7.0 of amount of the amount of liquid that sees through according to ultrafiltration continues reaction simultaneously.
After treating that the interior liquid of the 2nd grade of fixed enzyme membrane reactor cylinder reaches about 5L, open the 2nd grade of stirring rake, the beginning hydrolysis reaction, reaction conditions is identical with the 1st grade.Hydrolysis was opened ultra-filtration equipment after 1 hour approximately, and ultrafiltration pressure is 0.05Mpa, circular flow 10L/h.The phosphopeptide caseinate that the molecular weight that hydrolysis is produced is lower than 6000Da sees through, and is transformed into the outside of cylinder, and Open valve 6b, the phosphopeptide caseinate that makes molecular weight be lower than 6000Da flow in the 3rd level fixed enzyme membrane reactor cylinder.In the 2nd grade of fixed enzyme membrane reactor cylinder is the phosphopeptide caseinate of molecular weight between 6000～8000Da.
Directly open valve 6e, make from the outer liquid of the 2nd grade of fixed enzyme membrane reactor cylinder and directly inject nanofiltration device without hydrolysis reaction, open nanofiltration device, be used for concentrating and desalination of product, working pressure 0.15Mpa when dehydration rate reaches 50%, finishes nanofiltration.The nanofiltration concentrated solution is the phosphopeptide caseinate product of molecular weight at 350～6000Da.
Also but Open valve 6d directly injects nanofiltration device with liquid in the 2nd grade of fixed enzyme membrane reactor cylinder, obtains the phosphopeptide caseinate product of molecular weight at 6000～8000Da through the nanofiltration desalination and concentration.
After testing, the index of above-mentioned 2 kinds of phosphopeptide caseinates is as follows:
The phosphopeptide caseinate of molecular weight 6000～8000Da: purity: 24.21%, protein content: 93.37%, N content: P content 14.63%: 0.95%;
The phosphopeptide caseinate of molecular weight 350～6000Da: purity: 49.80%, protein content: 91.23%, N content: P content 14.30%: 1.43%.
Claims (10)
1. a multistage fixed enzyme membrane reaction unit is characterized in that it comprises the placed in-line fixed enzyme membrane reactor of N level and a nanofiltration device; Each grade fixed enzyme membrane reactor forms cylinder with the polysulfones porous material for being supported on inside reactor, be divided into reactor in the cylinder and outer two spaces of cylinder, strengthen the intensity of above-mentioned cylindrical structure with polyester non-woven fabric, with molecular weight cut-off is the inboard that the film of 1000～10000Da is seated in above-mentioned cylindrical structure, is fixed with proteolytic enzyme on the film inner surface; Be provided with the nanofiltration membrane that molecular weight cut-off is 100～500Da in the nanofiltration device; The 1st grade of fixed enzyme membrane reactor cylindrical outer is connected with the 2nd grade of fixed enzyme membrane reactor cylinder interior by pipeline, the 2nd grade of fixed enzyme membrane reactor cylindrical outer is connected with 3rd level fixed enzyme membrane reactor cylinder interior by pipeline, by that analogy, until N level fixed enzyme membrane reactor, N level fixed enzyme membrane reactor cylindrical outer is connected with nanofiltration device by pipeline, the 2nd grade is connected with nanofiltration device by pipeline respectively to N level fixed enzyme membrane reactor cylinder interior, and above-mentioned pipeline all is provided with valve; The 1st grade of fixed enzyme membrane reactor to the N level fixed enzyme membrane reactor, the molecular weight cut-off of its film successively decreases successively; Described N gets 2～100 integer.
2. multistage fixed enzyme membrane reaction unit according to claim 1 is characterized in that described molecular weight cut-off is that the film of 1000～10000Da is any one in polyamide composite film, cellulose acetate film and the polycarbonate composite membrane.
3. multistage fixed enzyme membrane reaction unit according to claim 1 is characterized in that described fixed enzyme membrane reactor is equipped with the temperature control chuck.
4. multistage fixed enzyme membrane reaction unit according to claim 1 is characterized in that described fixed enzyme membrane reactor cylinder interior is provided with stirring rake.
5. multistage fixed enzyme membrane reaction unit according to claim 1 is characterized in that described proteolytic enzyme is any one or a few in Sumizyme MP, pancreatin, papoid, neutral protease, the compound protease.
6. multistage fixed enzyme membrane reaction unit according to claim 1 is characterized in that described N gets 2～5 integer.
7. utilize the described multistage fixed enzyme membrane reaction unit of claim 1 to prepare the method for phosphopeptide caseinate, it is characterized in that casein solution is injected into the 1st grade of fixed enzyme membrane reactor cylinder interior, carry out enzyme digestion reaction, after reaction finishes, by ultrafiltration molecular weight in the cylinder interior liquid is transformed into cylindrical outer less than the material of the 1st grade of membrane molecule amount, by pipeline the material of the 1st grade of fixed enzyme membrane reactor cylindrical outer is injected the 2nd grade of fixed enzyme membrane reactor cylinder interior again, carry out enzyme digestion reaction, after reaction finishes, by ultrafiltration molecular weight in the cylinder interior liquid is transformed into cylindrical outer less than the material of the 2nd grade of membrane molecule amount, by that analogy, until N level fixed enzyme membrane reactor, N level fixed enzyme membrane reactor cylindrical outer liquid is injected nanofiltration device by pipeline, obtain molecular weight less than N level membrane molecule amount and greater than the phosphopeptide caseinate of nanofiltration membrane molecular weight through the nanofiltration desalination and concentration; Simultaneously can be after the 2nd grade of each grade fixed enzyme membrane reactor to the N level carries out ultrafiltration the material of this grade fixed enzyme membrane reactor cylinder interior be injected nanofiltration device, obtain molecular weight less than one-level membrane molecule amount before this grade and greater than the phosphopeptide caseinate of this grade membrane molecule amount, the 1st grade of fixed enzyme membrane reactor cylinder interior can be replenished the casein solution reaction repeated through the nanofiltration desalination and concentration.
8. the method for preparing phosphopeptide caseinate according to claim 7 is characterized in that described casein solution, and concentration is 40～150g/L, and pH is 6.5～9.5.
9. the method for preparing phosphopeptide caseinate according to claim 7, the temperature of reaction that it is characterized in that each grade fixed enzyme membrane reactor is 30～60 ℃, and stirring velocity is 50～250rpm, and the enzyme digestion reaction time is 1～3h.
10. the method for preparing phosphopeptide caseinate according to claim 7 is characterized in that each grade fixed enzyme membrane reactor ultrafiltration pressure is 0.01～0.08MPa.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201019026024.3A CN101768551A (en) | 2010-02-04 | 2010-02-04 | Multipole immobilized enzyme film reaction device and method of preparing casein phosphopeptides by using reaction device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201019026024.3A CN101768551A (en) | 2010-02-04 | 2010-02-04 | Multipole immobilized enzyme film reaction device and method of preparing casein phosphopeptides by using reaction device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101768551A true CN101768551A (en) | 2010-07-07 |
Family
ID=42501652
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201019026024.3A Pending CN101768551A (en) | 2010-02-04 | 2010-02-04 | Multipole immobilized enzyme film reaction device and method of preparing casein phosphopeptides by using reaction device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101768551A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103571905A (en) * | 2013-10-30 | 2014-02-12 | 广州绿萃生物科技有限公司 | Preparation method of high-purity casein phosphopeptide |
| CN103966292A (en) * | 2013-01-31 | 2014-08-06 | 中国食品发酵工业研究院 | Industrial manufacturing method and use of allergen-eliminated partly appropriately-hydrolyzed casein peptide |
| CN105420493A (en) * | 2015-12-28 | 2016-03-23 | 云南云铜锌业股份有限公司 | Wet-metallurgy continuous reaction kettle equipment |
| CN110804637A (en) * | 2019-11-27 | 2020-02-18 | 华熙生物科技股份有限公司 | Preparation method of active oligopeptide |
| CN111019820A (en) * | 2019-12-10 | 2020-04-17 | 沈阳农业大学 | Bioreactor and method for preparing pectin oligosaccharide and natural food preservative thereof |
| CN111225570A (en) * | 2017-09-08 | 2020-06-02 | 阿尔克雷斯塔治疗股份有限公司 | Apparatus and method for preparing and administering nutritional formulas |
| WO2020238319A1 (en) * | 2019-05-31 | 2020-12-03 | 基因港(香港)生物科技有限公司 | Immobilized enzyme reactor, and immobilized enzyme reaction device |
| CN112853559A (en) * | 2020-12-31 | 2021-05-28 | 盐城师范学院 | Process constraint nesting control system and method based on ring spinning yarn quality |
-
2010
- 2010-02-04 CN CN201019026024.3A patent/CN101768551A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103966292A (en) * | 2013-01-31 | 2014-08-06 | 中国食品发酵工业研究院 | Industrial manufacturing method and use of allergen-eliminated partly appropriately-hydrolyzed casein peptide |
| CN103571905A (en) * | 2013-10-30 | 2014-02-12 | 广州绿萃生物科技有限公司 | Preparation method of high-purity casein phosphopeptide |
| CN105420493A (en) * | 2015-12-28 | 2016-03-23 | 云南云铜锌业股份有限公司 | Wet-metallurgy continuous reaction kettle equipment |
| CN111225570A (en) * | 2017-09-08 | 2020-06-02 | 阿尔克雷斯塔治疗股份有限公司 | Apparatus and method for preparing and administering nutritional formulas |
| WO2020238319A1 (en) * | 2019-05-31 | 2020-12-03 | 基因港(香港)生物科技有限公司 | Immobilized enzyme reactor, and immobilized enzyme reaction device |
| CN110804637A (en) * | 2019-11-27 | 2020-02-18 | 华熙生物科技股份有限公司 | Preparation method of active oligopeptide |
| CN110804637B (en) * | 2019-11-27 | 2021-09-10 | 华熙生物科技股份有限公司 | Preparation method of active oligopeptide |
| CN111019820A (en) * | 2019-12-10 | 2020-04-17 | 沈阳农业大学 | Bioreactor and method for preparing pectin oligosaccharide and natural food preservative thereof |
| CN111019820B (en) * | 2019-12-10 | 2022-11-25 | 沈阳农业大学 | Bioreactor and method for preparing pectin oligosaccharide and natural food preservative thereof |
| CN112853559A (en) * | 2020-12-31 | 2021-05-28 | 盐城师范学院 | Process constraint nesting control system and method based on ring spinning yarn quality |
| CN112853559B (en) * | 2020-12-31 | 2021-10-08 | 盐城师范学院 | Process constraint nesting control system and method based on ring spinning yarn quality |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101768551A (en) | Multipole immobilized enzyme film reaction device and method of preparing casein phosphopeptides by using reaction device | |
| CN103571905B (en) | Preparation method of high-purity casein phosphopeptide | |
| CN102329846B (en) | Pollution-free production method for producing soybean and silkworm chrysalis composite oligopeptides by adopting composite enzyme hydrolysis method | |
| CN103045706B (en) | Method for continuously producing scale collagen peptide chelated calcium salt and scale collagen peptide | |
| CN101824085B (en) | Method for separating alpha s-casein | |
| CN103045707B (en) | Method for preparing rice protein active polypeptide through continuous enzymolysis and secondary membrance filtering and purification | |
| CN103352064A (en) | Method for preparing corn protein active peptide by using composite carrier immobilized double enzymes | |
| CN102051130A (en) | Method for preparing gelatin with protease degradation ossein | |
| CN102329843A (en) | Enzyme method for preparing gelatin | |
| CN102229643A (en) | Method for preparing high-purity rice protein and high-purity rice peptide | |
| CN101643766B (en) | Method for preparing hydrolyzed collagen protein by fresh animal skin | |
| CN109201704A (en) | Vacuum drum filters tailings recycling treatment and Zero discharging system and method | |
| CN103739663B (en) | Microwave-assisted acid hydrolysis prepares the method for little peptide ammino acid fast | |
| CN101838467B (en) | Novel chitosan nanoparticles and preparation method thereof | |
| CN101849605A (en) | Preparation method of silkworm pupa high zinc protein powder | |
| CN103320487A (en) | Method for preparing trepang peptide by combined hydrolysis of soluble enzyme and immobilized enzyme | |
| CN103421868A (en) | Preparation method of tuna skin collagen micro-molecular peptides | |
| CN103421871A (en) | Preparation method of tuna bone collagen peptide | |
| CN102174503A (en) | Spray drying micro-capsulation process for high-yield alkaline protease | |
| CN106220521A (en) | A kind of full film extracts the method for L isoleucine | |
| CN103103243A (en) | Method for preparing silkworm chrysalis peptide by conducting combined hydrolysis on free enzyme and immobilized enzyme | |
| CN102150741B (en) | Method for preparing lactalbumin hydrolysate by utilizing yak milk | |
| CN108504645A (en) | A kind of preparation method of Pullulanase | |
| CN105154505B (en) | A kind of preparation method of feed grade ocean fish oligopeptide powder | |
| CN105154506B (en) | A kind of food-grade low salt ocean fish oligopeptide powder and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20100707 |