CN106591408A - Method for preparing trepang collagen peptide by adopting microbiological fermentation method - Google Patents
Method for preparing trepang collagen peptide by adopting microbiological fermentation method Download PDFInfo
- Publication number
- CN106591408A CN106591408A CN201611227655.8A CN201611227655A CN106591408A CN 106591408 A CN106591408 A CN 106591408A CN 201611227655 A CN201611227655 A CN 201611227655A CN 106591408 A CN106591408 A CN 106591408A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- collagen peptide
- parts
- bacillus
- trepang
- 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
- 238000000855 fermentation Methods 0.000 title claims abstract description 62
- 230000004151 fermentation Effects 0.000 title claims abstract description 62
- 102000008186 Collagen Human genes 0.000 title claims abstract description 49
- 108010035532 Collagen Proteins 0.000 title claims abstract description 49
- 238000000034 method Methods 0.000 title claims abstract description 42
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 41
- 229920001436 collagen Polymers 0.000 title claims abstract description 33
- 230000002906 microbiologic effect Effects 0.000 title abstract 2
- 238000004108 freeze drying Methods 0.000 claims abstract description 5
- 241000251511 Holothuroidea Species 0.000 claims description 35
- 239000007788 liquid Substances 0.000 claims description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 25
- 239000000706 filtrate Substances 0.000 claims description 22
- 239000002994 raw material Substances 0.000 claims description 17
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 12
- 230000008569 process Effects 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- 239000002002 slurry Substances 0.000 claims description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 9
- 239000008103 glucose Substances 0.000 claims description 9
- 239000001963 growth medium Substances 0.000 claims description 9
- 239000012528 membrane Substances 0.000 claims description 9
- 238000011218 seed culture Methods 0.000 claims description 9
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 6
- 241000194108 Bacillus licheniformis Species 0.000 claims description 6
- 239000002361 compost Substances 0.000 claims description 6
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 5
- 241000194107 Bacillus megaterium Species 0.000 claims description 5
- 241000628997 Flos Species 0.000 claims description 5
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims description 5
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 claims description 5
- 238000010009 beating Methods 0.000 claims description 5
- 235000015278 beef Nutrition 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 5
- 239000000084 colloidal system Substances 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 5
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- 238000007654 immersion Methods 0.000 claims description 5
- 239000002054 inoculum Substances 0.000 claims description 5
- 229930182817 methionine Natural products 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 238000007789 sealing Methods 0.000 claims description 5
- 229910052711 selenium Inorganic materials 0.000 claims description 5
- 239000011669 selenium Substances 0.000 claims description 5
- 235000010413 sodium alginate Nutrition 0.000 claims description 5
- 239000000661 sodium alginate Substances 0.000 claims description 5
- 229940005550 sodium alginate Drugs 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 241000193755 Bacillus cereus Species 0.000 claims description 4
- 229920002774 Maltodextrin Polymers 0.000 claims description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 4
- 239000012530 fluid Substances 0.000 claims description 4
- 230000014759 maintenance of location Effects 0.000 claims description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 4
- 108091005804 Peptidases Proteins 0.000 claims description 3
- 239000004365 Protease Substances 0.000 claims description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- 238000000889 atomisation Methods 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 2
- 150000003625 trehaloses Chemical class 0.000 claims description 2
- -1 acyl Amine Chemical class 0.000 claims 1
- 238000010025 steaming Methods 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 238000002360 preparation method Methods 0.000 abstract description 3
- 235000019658 bitter taste Nutrition 0.000 abstract description 2
- 238000001914 filtration Methods 0.000 abstract 1
- 230000003647 oxidation Effects 0.000 abstract 1
- 238000007254 oxidation reaction Methods 0.000 abstract 1
- 238000004537 pulping Methods 0.000 abstract 1
- 238000002791 soaking Methods 0.000 abstract 1
- 238000001694 spray drying Methods 0.000 abstract 1
- 241000965254 Apostichopus japonicus Species 0.000 description 13
- 102000004196 processed proteins & peptides Human genes 0.000 description 7
- 229920001184 polypeptide Polymers 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 235000001014 amino acid Nutrition 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 3
- 102000002322 Egg Proteins Human genes 0.000 description 3
- 108010000912 Egg Proteins Proteins 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 235000014103 egg white Nutrition 0.000 description 3
- 210000000969 egg white Anatomy 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000000227 grinding Methods 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 239000005913 Maltodextrin Substances 0.000 description 2
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 229940035034 maltodextrin Drugs 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 2
- 229930182490 saponin Natural products 0.000 description 2
- 150000007949 saponins Chemical class 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 235000019750 Crude protein Nutrition 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000010874 maintenance of protein location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229940126578 oral vaccine Drugs 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000037394 skin elasticity Effects 0.000 description 1
- 239000012879 subculture medium Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method for preparing trepang collagen peptide by adopting a microbiological fermentation method. The method provided by the invention comprises the following steps: soaking East China Sea black trepang, carrying out pulping, carrying out enzymolysis, carrying out filtering, inoculating the to-be-fermented seeds, and after fermentation, carrying out freeze drying or spray drying, thus obtaining trepang collagen peptide. The preparation method provided by the invention has the advantages that the production cycle is short, the yield is high, the product is high in purity, has the obvious oxidation resistance, is good in solubility, high in absorbency, and free of foreign smell or bitter taste, and in addition, trepang is sufficiently processed and utilized for high values.
Description
Technical field
The present invention relates to a kind of preparation of collagen peptide, more particularly to sea cucumber collagen prepared by a kind of microbe fermentation method
The method of protein peptide.
Background technology
Collagen polypeptide is that collagen or gelatin are obtained between protein and aminoacid Jing after the degradation treatments such as protease
Product, typically will be referred to as oligopeptide or small peptide by the linear peptides of peptide bond formed by 2 to 10 aminoacid；Got up by peptide linkage
The polymer more than 10 aminoacid then be referred to as polypeptide.In recent years, many scholars are ground to the mechanism of digesting and assimilating of peptide
Study carefully, it is believed that peptide even considerable amount of protein can be utilized in animal body by absorbed intact and by body, such as oral vaccine and
Enzyme process treats disease.The movement system of peptide compares transport speed with free amino acid faster.Many tests show that the addition of peptide is also
The infiltration rate of aminoacid can be improved.In addition peptide can also promote Protein retention, because peptide can be directly used in human body protein
The synthesis of matter.Long-term consumption contributes to improving skin elasticity through the food of addition collagen peptide, and slow down aging strengthens immunity
Power, while instant, is suitable for general population.
Stichopus japonicuss are important marine foods and drug resource.There are more than 1000 kinds of Stichopus japonicuss in the whole world, wherein kind more than 40 is available for food
With.In recent years, China's holothruian cultures industry development is exceedingly fast, and Stichopus japonicuss annual production is more than 60,000 tons.It would therefore be highly desirable to further exploitation is extra large
The high-valued process technology of ginseng.Wall of sea cucumber Stichopus japonicus contains active substance such as holothurian collagen, Stichopus japonicus polysaccharide, the Stichopus japonicuss of unique structure
Saponin etc..Pharmacological evaluation proves that Stichopus japonicuss have antitumor, improve immunologic function, blood fat reducing, anticoagulation, atherosclerosiss
Etc. various physiologically actives.At present, both at home and abroad Stichopus japonicus polysaccharide and Stichopus japonicuss are concentrated mainly on to the research and development of Stichopus japonicuss active component
The aspects such as saponin.
Holothurian collagen content accounts for more than the 60% of wall of sea cucumber Stichopus japonicus crude protein, about holothurian collagen type and physics and chemistry
Property Quality Research have been reported (Comp Biochem Physiol, 1995,112A：463-478；Food Chemistry
AndToxicology, 2002,67 (4)：1319-132；Aquatic product journal, 2006,4：549-553), Chinese invention patent CN
101215315A also discloses that a kind of (0-25 DEG C) method for extracting holothurian collagen using pepsin dissolution at low temperature.
Compared with collagen protein, collagen peptide has the features such as being more easy to absorption and high physiologically active.About sea cucumber polypeptide research
Have been reported that, at present published patent of invention there are CN 1858222A and CN 101191139A.Stichopus japonicuss described in these researchs
Polypeptide is a kind of at least including the mixture of selenka, protein polypeptide and collagen polypeptide isoreactivity material.Relevant sea cucumber collagen
There is not been reported for protein active peptide.
The content of the invention
The invention provides a kind of improve purity and extraction ratio, remove bitter off-flavors, improve mouthfeel, beneficial to the micro- of absorption of human body
The method of holothurian collagen peptide prepared by biological fermentation process.
The present invention is by the following technical programs：
A kind of method of holothurian collagen peptide prepared by microbe fermentation method, comprises the technical steps that：
A. it is raw material to choose the black sea cucumbers from East China Sea rich in collagen protein, and clear water immersion removing salinity immerses soak 50- after cleaning
60min, pulls out and drains away the water, and using beater beating process is carried out, and adds and adopt after the pure water of 35-40 times of volume colloid mill
Micronization processes are ground, slurry is obtained；
B. the slurry that step a is obtained is heated to into central temperature and reaches 130-145 DEG C, keep 10min, be cooled to 20-25 DEG C,
Add protease to be digested, cross 120 mesh filter screens, obtain collagen protein filtrate；
C. in the collagen protein filtrate that step b is obtained, weight percent 0.5-3% adds glucose or sucrose, adjusts pH
To 6-7, Preliminary fermentation liquid is obtained；
D. the Preliminary fermentation liquid for step c being obtained be put into sky disappear fermentation tank carry out it is real disappear, sky disappears and the real condition for disappearing is 121
DEG C, 20min, treat tank temperature drop to 37 DEG C, access fermentation seed, inoculum concentration is the 2-4.5% of Preliminary fermentation liquid percentage by volume, is connect
Mixed liquor sealing and fermenting 2-5 days under conditions of 37 DEG C and 100-300rpm speeds of agitator is planted, fermentation liquid filters off residue, must ferment
Filtrate；
E. 15-20% trehaloses, 0.3-0.8% maltodextrins and 3-5% glycerol are added in the ferment filtrate that step d is obtained ,-
50 DEG C of pre-freeze 2-6h, then the lyophilization 12-20h under conditions of -70 DEG C of condenser temperatures and ＜ 150Pa vacuums, directly obtains
To holothurian collagen peptide；Or, the ferment filtrate that step d is obtained adopts aperture and divides for 0.1-0.5 μm of tubular membrane removing is big
Son, then adopts molecular cut off and is classified for the ultrafilter membrane retention of 3kDa, and trapped fluid adopts NF membrane concentrating and desalinating, Ran Hou
It is spray-dried under conditions of 175-185 DEG C of inlet temperature, 75-85 DEG C of leaving air temp and 0.35-0.45MPa atomisation pressures, obtains
Holothurian collagen peptide.
Preferably, soak is made up of the raw material of following weight portion in step a：Fructus Crataegi 20-25 parts, Flos Chrysanthemi Indici 1-5 parts,
The matter of Herba Pogostemonis 30-40 parts, shitosan 15-18 parts, sodium alginate 12-15 parts and water 200-350 parts, black sea cucumbers from East China Sea and soak
Amount is than being 1:10.
Preferably, fermentation seed is compost ground bacillus, Bacillus licheniformis or huge spore bar on the spot in step d
Bacterium.
Preferably, the raw material of fermentation seed culture medium is based on following parts by weight：3% soy peptone, 5% selenium-rich ferment
Mother, 6% agedoite, 5.5% methionine, 0.5% beef powder, 0.5% glucose, 0.1% Sodium Chloride, 0.01% biphosphate
Potassium, remaining is distilled water, and 121 DEG C of high temperature sterilize 20min are standby after cooling.
Preferably, compost ground bacillus, Bacillus licheniformis or bacillus megaterium fermentation kind will be inoculated on the spot
In sub- culture medium, shaking table 180r/min cultivates 48h, bacillus cereuss ＞ 10 in fermentation liquid8cfu/mL。
The invention has the beneficial effects as follows：The preparation method of the present invention is with short production cycle, and yield is high, and product purity is high, has
Significant non-oxidizability, solubility is good, and absorbability is high, free from extraneous odour and bitterness；Stichopus japonicuss are made to obtain sufficient high-valued processing and utilization.
Specific embodiment
To make those skilled in the art more fully understand technical scheme, but these elaborations are not done to the present invention
Any pro forma restriction.Unless otherwise indicated, all scientific and technical terms otherwise used by the present invention have belonging to the present invention
The implication being generally understood with those possessing an ordinary skill in the pertinent arts.
Further details of elaboration is made to the present invention with reference to specific embodiment, it is to be understood that, embodiment should not
It is interpreted as limiting the scope of the invention.
Embodiment 1
A kind of method of holothurian collagen peptide prepared by microbe fermentation method, comprises the technical steps that：
A. it is raw material to choose the black sea cucumbers from East China Sea rich in collagen protein, and clear water immersion removing salinity immerses soak after cleaning
50min, pulls out and drains away the water, and using beater beating process is carried out, and is entered using colloid mill after the pure water for adding 35 times of volumes
Row grinding micronization processes, obtain slurry；Soak is made up of the raw material of following weight portion：20 parts of Fructus Crataegi, 1 part of Flos Chrysanthemi Indici, the wide leaves of pulse plants
Fragrant 30 parts, 15 parts of shitosan, 12 parts of sodium alginate are 1 with the mass ratio of 200 parts of water, black sea cucumbers from East China Sea and soak:10；
B. the slurry that step a is obtained is heated to into central temperature and reaches 130 DEG C, keep 10min, be cooled to 20 DEG C, add egg
White enzyme is digested, and crosses 120 mesh filter screens, obtains collagen protein filtrate；
C. in the collagen protein filtrate that step b is obtained, weight percent 0.5% adds glucose, adjusts pH to 6, obtains just
Originate zymotic fluid；
D. the Preliminary fermentation liquid for step c being obtained be put into sky disappear fermentation tank carry out it is real disappear, sky disappears and the real condition for disappearing is 121
DEG C, 20min, treat tank temperature drop to 37 DEG C, access fermentation seed, inoculum concentration is the 2% of Preliminary fermentation liquid percentage by volume, and inoculation is mixed
Liquid sealing and fermenting 2 days under conditions of 37 DEG C and 100rpm speeds of agitator are closed, fermentation liquid filters off residue, obtains ferment filtrate；Fermentation
Seed is compost ground bacillus on the spot；The raw material of fermentation seed culture medium is based on following parts by weight：3% soy peptone,
5% yeast rich in selenium, 6% agedoite, 5.5% methionine, 0.5% beef powder, 0.5% glucose, 0.1% Sodium Chloride,
0.01% potassium dihydrogen phosphate, remaining is distilled water, and 121 DEG C of high temperature sterilize 20min are standby after cooling；
Compost ground bacillus will be inoculated in fermentation seed culture medium on the spot, shaking table 180r/min cultivates 48h, in fermentation liquid
Bacillus cereuss ＞ 108cfu/mL；
E. 15% trehalose, 0.3% maltodextrin and 3% glycerol, -50 DEG C of pre-freezes are added in the ferment filtrate that step d is obtained
2h, then the lyophilization 12h under conditions of -70 DEG C of condenser temperatures and ＜ 150Pa vacuums, directly obtains holothurian collagen
Peptide.
Embodiment 2
A kind of method of holothurian collagen peptide prepared by microbe fermentation method, comprises the technical steps that：
A. it is raw material to choose the black sea cucumbers from East China Sea rich in collagen protein, and clear water immersion removing salinity immerses soak after cleaning
55min, pulls out and drains away the water, and using beater beating process is carried out, and is entered using colloid mill after the pure water for adding 38 times of volumes
Row grinding micronization processes, obtain slurry；Soak is made up of the raw material of following weight portion：22 parts of Fructus Crataegi, 3 parts of Flos Chrysanthemi Indici, the wide leaves of pulse plants
Fragrant 36 parts, 17 parts of shitosan, 13 parts of sodium alginate are 1 with the mass ratio of 250 parts of water, black sea cucumbers from East China Sea and soak:10；
B. the slurry that step a is obtained is heated to into central temperature and reaches 142 DEG C, keep 10min, be cooled to 22 DEG C, add egg
White enzyme is digested, and crosses 120 mesh filter screens, obtains collagen protein filtrate；
C. in the collagen protein filtrate that step b is obtained, weight percent 2% adds sucrose, adjusts pH to 6.5, obtains initially
Fermentation liquid；
D. the Preliminary fermentation liquid for step c being obtained be put into sky disappear fermentation tank carry out it is real disappear, sky disappears and the real condition for disappearing is 121
DEG C, 20min, treat tank temperature drop to 37 DEG C, access fermentation seed, inoculum concentration is the 3.2% of Preliminary fermentation liquid percentage by volume, inoculation
Mixed liquor sealing and fermenting 3 days under conditions of 37 DEG C and 200rpm speeds of agitator, fermentation liquid filters off residue, obtains ferment filtrate；Send out
Ferment seed is Bacillus licheniformis；The raw material of fermentation seed culture medium is based on following parts by weight：It is 3% soy peptone, 5% rich
Selenium yeast, 6% agedoite, 5.5% methionine, 0.5% beef powder, 0.5% glucose, 0.1% Sodium Chloride, 0.01% phosphoric acid
Potassium dihydrogen, remaining is distilled water, and 121 DEG C of high temperature sterilize 20min are standby after cooling；
By Bacillus licheniformis in fermentation seed culture medium, shaking table 180r/min cultivates 48h, bacillus cereuss ＞ in fermentation liquid
108cfu/mL；
E. the ferment filtrate that step d is obtained adopts aperture and removes macromole for 0.1-0.5 μm of tubular membrane, then using retention point
Son amount for 3kDa ultrafilter membrane retention classification, trapped fluid adopt NF membrane concentrating and desalinating, then 175 DEG C of inlet temperature, 75 DEG C
It is spray-dried under conditions of leaving air temp and 0.35MPa atomisation pressures, obtains holothurian collagen peptide.
Embodiment 3
A kind of method of holothurian collagen peptide prepared by microbe fermentation method, comprises the technical steps that：
A. it is raw material to choose the black sea cucumbers from East China Sea rich in collagen protein, and clear water immersion removing salinity immerses soak after cleaning
60min, pulls out and drains away the water, and using beater beating process is carried out, and is entered using colloid mill after the pure water for adding 40 times of volumes
Row grinding micronization processes, obtain slurry；Soak is made up of the raw material of following weight portion：25 parts of Fructus Crataegi, 5 parts of Flos Chrysanthemi Indici, the wide leaves of pulse plants
Fragrant 40 parts, 18 parts of shitosan, 15 parts of sodium alginate are 1 with the mass ratio of 350 parts of water, black sea cucumbers from East China Sea and soak:10；
B. the slurry that step a is obtained is heated to into central temperature and reaches 145 DEG C, keep 10min, be cooled to 25 DEG C, add egg
White enzyme is digested, and crosses 120 mesh filter screens, obtains collagen protein filtrate；
C. in the collagen protein filtrate that step b is obtained, weight percent 3% adds glucose, adjusts pH to 7, obtains initially
Fermentation liquid；
D. the Preliminary fermentation liquid for step c being obtained be put into sky disappear fermentation tank carry out it is real disappear, sky disappears and the real condition for disappearing is 121
DEG C, 20min, treat tank temperature drop to 37 DEG C, access fermentation seed, inoculum concentration is the 4.5% of Preliminary fermentation liquid percentage by volume, inoculation
Mixed liquor sealing and fermenting 5 days under conditions of 37 DEG C and 300rpm speeds of agitator, fermentation liquid filters off residue, obtains ferment filtrate；Send out
Ferment seed is bacillus megaterium；The raw material of fermentation seed culture medium is based on following parts by weight：It is 3% soy peptone, 5% rich
Selenium yeast, 6% agedoite, 5.5% methionine, 0.5% beef powder, 0.5% glucose, 0.1% Sodium Chloride, 0.01% phosphoric acid
Potassium dihydrogen, remaining is distilled water, and 121 DEG C of high temperature sterilize 20min are standby after cooling；
Bacillus megaterium is inoculated in fermentation seed culture medium, shaking table 180r/min, cultivates 48h, spore bar in fermentation liquid
Bacterium ＞ 108cfu/mL；
E. 20% trehalose, 0.8% maltodextrin and 5% glycerol, -50 DEG C of pre-freezes are added in the ferment filtrate that step d is obtained
6h, then the lyophilization 20h under conditions of -70 DEG C of condenser temperatures and ＜ 150Pa vacuums, directly obtains holothurian collagen
Peptide.
Raw materials used in the present invention, equipment, unless otherwise noted, is the conventional raw material, equipment of this area；In the present invention
Method therefor, unless otherwise noted, is the conventional method of this area.
The above, is only presently preferred embodiments of the present invention, and not the present invention is imposed any restrictions, every according to the present invention
Any simple modification, change and equivalent transformation that technical spirit is made to above example, still fall within the technology of the present invention side
The protection domain of case.
Claims (5)
1. the method for holothurian collagen peptide prepared by a kind of microbe fermentation method, it is characterised in that comprise the technical steps that：
A. it is raw material to choose the black sea cucumbers from East China Sea rich in collagen protein, and clear water immersion removing salinity immerses soak 50- after cleaning
60min, pulls out and drains away the water, and using beater beating process is carried out, and adds and adopt after the pure water of 35-40 times of volume colloid mill
Micronization processes are ground, slurry is obtained；
B. the slurry that step a is obtained is heated to into central temperature and reaches 130-145 DEG C, keep 10min, be cooled to 20-25 DEG C,
Add protease to be digested, cross 120 mesh filter screens, obtain collagen protein filtrate；
C. in the collagen protein filtrate that step b is obtained, weight percent 0.5-3% adds glucose or sucrose, adjusts pH
To 6-7, Preliminary fermentation liquid is obtained；
D. the Preliminary fermentation liquid for step c being obtained be put into sky disappear fermentation tank carry out it is real disappear, sky disappears and the real condition for disappearing is 121
DEG C, 20min, treat tank temperature drop to 37 DEG C, access fermentation seed, inoculum concentration is the 2-4.5% of Preliminary fermentation liquid percentage by volume, is connect
Mixed liquor sealing and fermenting 2-5 days under conditions of 37 DEG C and 100-300rpm speeds of agitator is planted, fermentation liquid filters off residue, must ferment
Filtrate；
E. 15-20% trehaloses, 0.3-0.8% maltodextrins and 3-5% glycerol are added in the ferment filtrate that step d is obtained ,-
50 DEG C of pre-freeze 2-6h, then the lyophilization 12-20h under conditions of -70 DEG C of condenser temperatures and ＜ 150Pa vacuums, directly obtains
To holothurian collagen peptide；Or, the ferment filtrate that step d is obtained adopts aperture and divides for 0.1-0.5 μm of tubular membrane removing is big
Son, then adopts molecular cut off and is classified for the ultrafilter membrane retention of 3kDa, and trapped fluid adopts NF membrane concentrating and desalinating, Ran Hou
It is spray-dried under conditions of 175-185 DEG C of inlet temperature, 75-85 DEG C of leaving air temp and 0.35-0.45MPa atomisation pressures, obtains
Holothurian collagen peptide.
2. the method for holothurian collagen peptide prepared by a kind of microbe fermentation method according to claim 1, its feature exists
In soak is made up of the raw material of following weight portion in step a：Fructus Crataegi 20-25 parts, Flos Chrysanthemi Indici 1-5 parts, Herba Pogostemonis 30-40 parts,
Shitosan 15-18 parts, sodium alginate 12-15 parts and water 200-350 parts, black sea cucumbers from East China Sea is 1 with the mass ratio of soak:10.
3. the method for holothurian collagen peptide prepared by a kind of microbe fermentation method according to claim 1, its feature exists
In fermentation seed is compost ground bacillus, Bacillus licheniformis or bacillus megaterium on the spot in step d.
4. the method for holothurian collagen peptide prepared by a kind of microbe fermentation method according to claim 3, its feature exists
In the raw material of fermentation seed culture medium is based on following parts by weight：3% soy peptone, 5% yeast rich in selenium, 6% Radix Asparagi acyl
Amine, 5.5% methionine, 0.5% beef powder, 0.5% glucose, 0.1% Sodium Chloride, 0.01% potassium dihydrogen phosphate, remaining is steaming
Distilled water, 121 DEG C of high temperature sterilize 20min are standby after cooling.
5. the method for holothurian collagen peptide prepared by a kind of microbe fermentation method according to claim 4, its feature exists
In compost ground bacillus, Bacillus licheniformis or bacillus megaterium being inoculated in fermentation seed culture medium on the spot, shaking table
180r/min, cultivates 48h, bacillus cereuss ＞ 10 in fermentation liquid8cfu/mL。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611227655.8A CN106591408A (en) | 2016-12-27 | 2016-12-27 | Method for preparing trepang collagen peptide by adopting microbiological fermentation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611227655.8A CN106591408A (en) | 2016-12-27 | 2016-12-27 | Method for preparing trepang collagen peptide by adopting microbiological fermentation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106591408A true CN106591408A (en) | 2017-04-26 |
Family
ID=58603902
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611227655.8A Pending CN106591408A (en) | 2016-12-27 | 2016-12-27 | Method for preparing trepang collagen peptide by adopting microbiological fermentation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106591408A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107198673A (en) * | 2017-05-26 | 2017-09-26 | 山东姣梨坊生物科技有限公司 | Marine organisms care compositions and its production method |
| CN107549682A (en) * | 2017-09-28 | 2018-01-09 | 句容市武杭传动机械制造有限公司 | The processing method of a fermentation cucumber product |
| CN108220372A (en) * | 2018-01-16 | 2018-06-29 | 浙江海洋大学 | It is a kind of to prepare collagen from black sea cucumbers from East China Sea anti-oxidation peptide method with microbial protease enzymolysis |
| CN108410934A (en) * | 2018-03-19 | 2018-08-17 | 温州科技职业学院 | A kind of preparation method of acaudina molpadioidea collagen oligopeptide |
| CN109259215A (en) * | 2018-10-08 | 2019-01-25 | 北京国康本草物种生物科学技术研究院有限公司 | A kind of active material of sea cucumber and its separating and extracting process |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102251006A (en) * | 2011-07-29 | 2011-11-23 | 宁波超星海洋生物制品有限公司 | Method for manufacturing collagen from black sea cucumbers from East China Sea |
| CN102649974A (en) * | 2012-05-14 | 2012-08-29 | 上海海洋大学 | Method for preparing antioxidant peptide through ultrasonic-microwave synergetic enzymatic hydrolysis |
| CN103184262A (en) * | 2013-03-22 | 2013-07-03 | 浙江工业大学 | Method for extracting and purifying pepsin-soluble collagens of black sea cucumber in East China Sea |
| CN104878062A (en) * | 2015-05-18 | 2015-09-02 | 华南理工大学 | Microbiological method for preparing sea-fish protein antihypertensive peptides |
-
2016
- 2016-12-27 CN CN201611227655.8A patent/CN106591408A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102251006A (en) * | 2011-07-29 | 2011-11-23 | 宁波超星海洋生物制品有限公司 | Method for manufacturing collagen from black sea cucumbers from East China Sea |
| CN102649974A (en) * | 2012-05-14 | 2012-08-29 | 上海海洋大学 | Method for preparing antioxidant peptide through ultrasonic-microwave synergetic enzymatic hydrolysis |
| CN103184262A (en) * | 2013-03-22 | 2013-07-03 | 浙江工业大学 | Method for extracting and purifying pepsin-soluble collagens of black sea cucumber in East China Sea |
| CN104878062A (en) * | 2015-05-18 | 2015-09-02 | 华南理工大学 | Microbiological method for preparing sea-fish protein antihypertensive peptides |
Non-Patent Citations (1)
| Title |
|---|
| 童静静: "东海乌参蛋白肽的酶法制取及精制研究", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107198673A (en) * | 2017-05-26 | 2017-09-26 | 山东姣梨坊生物科技有限公司 | Marine organisms care compositions and its production method |
| CN107549682A (en) * | 2017-09-28 | 2018-01-09 | 句容市武杭传动机械制造有限公司 | The processing method of a fermentation cucumber product |
| CN108220372A (en) * | 2018-01-16 | 2018-06-29 | 浙江海洋大学 | It is a kind of to prepare collagen from black sea cucumbers from East China Sea anti-oxidation peptide method with microbial protease enzymolysis |
| CN108410934A (en) * | 2018-03-19 | 2018-08-17 | 温州科技职业学院 | A kind of preparation method of acaudina molpadioidea collagen oligopeptide |
| CN109259215A (en) * | 2018-10-08 | 2019-01-25 | 北京国康本草物种生物科学技术研究院有限公司 | A kind of active material of sea cucumber and its separating and extracting process |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106591408A (en) | Method for preparing trepang collagen peptide by adopting microbiological fermentation method | |
| CN101623065B (en) | Preparation method of fermented rice sticks | |
| CN102924423B (en) | Method for extracting cyanidin from banana peel | |
| CN106755240A (en) | A kind of method that microwave/ultrasonic assistant, microprotein enzyme hydrolysis prepare crow ginseng collagen oligopeptide | |
| CN105249062B (en) | A kind of plateau plant ferment drink and preparation method thereof | |
| CN102876559B (en) | Healthcare kelp vinegar and preparation method thereof | |
| CN105410873B (en) | A kind of functional kelp sauce rich in dietary fiber and preparation method thereof | |
| CN106916861B (en) | Method for simultaneously producing auricularia auricula polysaccharide and melanin | |
| CN109315656A (en) | A kind of preparation method of multi-cultur es composite fermentation rose ferment | |
| CN109957040A (en) | A kind of method that tremella polysaccharides are extracted in the mashing of enzyme process Aided Wet | |
| CN104757682A (en) | Method for preparing clear ginkgo fruit juice lactic acid beverage by using lactic acid bacteria | |
| CN105231401A (en) | Composite dried collybia albuminosa sauce and preparation method thereof | |
| CN104387488A (en) | Method for extracting mesona chinensis benth polysaccharide through microbial fermentation | |
| CN111543254A (en) | Culture method of phellinus igniarius mycelium, drink and application of phellinus igniarius mycelium | |
| CN102344872A (en) | Method for preparing sweet potato yellow wine containing anthocyanidin | |
| CN110236060B (en) | Mushroom solid instant beverage containing active lactic acid bacteria and preparation method thereof | |
| CN107372807A (en) | A kind of agent keeping vegetable fresh and preparation method thereof | |
| CN113249359B (en) | Method for extracting mulberry leaf functional ingredient by utilizing enzyme | |
| CN114214151A (en) | Preparation method of flower-flavor plum wine | |
| CN107841410A (en) | A kind of emblic fermented wine and preparation method thereof | |
| CN106539075A (en) | A kind of preparation method of efficient absorption sea cucumber nutrient buccal tablet | |
| CN105918547B (en) | A kind of agaricus bisporus sea-buckthorn instant tea and preparation method thereof | |
| AU2020102037A4 (en) | A method of efficiently increasing the alpha-glucosidase inhibitor content in fresh mulberry leaves by the solid-state fermentation | |
| CN111838354A (en) | Method for increasing selenium content in instant tea | |
| CN109837199A (en) | A kind of Spondias axillaris fruit vinegar and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170426 |