CN107177657A - A kind of method that step fermentation soft-shelled turtle egg prepares anti-oxidation peptide liquid - Google Patents
A kind of method that step fermentation soft-shelled turtle egg prepares anti-oxidation peptide liquid Download PDFInfo
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- CN107177657A CN107177657A CN201710529601.5A CN201710529601A CN107177657A CN 107177657 A CN107177657 A CN 107177657A CN 201710529601 A CN201710529601 A CN 201710529601A CN 107177657 A CN107177657 A CN 107177657A
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Abstract
The invention discloses a kind of method that step fermentation soft-shelled turtle egg prepares anti-oxidation peptide liquid.While the present invention crushes soft-shelled turtle egg progress liquid agitation, egg shell membrane this good anti-oxidation peptide raw material for remaining that common scrambled egg mode abandons, obtained soft-shelled turtle egg mixed liquor, substep is fermented with bacillus subtilis and lactic acid bacteria, each zymogenic advantage is preferably played, the anti-oxidation peptide fixture of preparation has more preferable inoxidizability.
Description
Technical field
The present invention relates to biological technical field, and in particular to a kind of step fermentation soft-shelled turtle egg prepares the side of anti-oxidation peptide liquid
Method.
Background technology
The cultivation of soft-shelled turtle has higher economic value in Chinese market, and soft-shelled turtle egg is removed as the accessory substance of breeding soft-shell turtle
Small part hatchery can sell fertilization egg incubation into soft-shelled turtle seedling, and it is former that most soft-shelled turtle egg is all only used as ordinary food
Material processing is even dropped, and causes certain wasting of resources.
The normal physiological metabolism of human body can produce some active oxygen radicals, and these free radicals can destroy protein, fat
Fat, nucleic acid etc., cause cell damage even apoptosis, human body are caused damage.And antioxidant activity peptide can be by freedom
Base provides proton or electronics to suppress free chain reaction progress, remove free radical, anti-lipid peroxidation, or chelant metal
The different approaches such as ion play antioxidation.
At present, the acquisition modes of anti-oxidation peptide mainly have the modes such as chemical synthesis, protease hydrolyzed, fermentation.Xu Huaide etc.
People carries out enzyme hydrolysis to turtle proteins and has obtained antioxidation active peptides；Yang Ming firm et al. applications are " a kind of that soft-shelled turtle egg is carried out into enzyme
The method that solution hydrolysis prepares antioxidation activity liquid " CN201510150779.X patent, has certain breakthrough in this field
Property.But the method has that cost is high, hydrolysising condition requirement is strict, there is enzymolysis product that is producing unpleasant bitter taste etc. to lack
Fall into, limit the application of anti-oxidation peptide.
Fermentation method is the microorganism of access production protease, using in fermentation process using soft-shelled turtle egg liquid as zymotic fluid
The protease of generation is by proteolysis into anti-oxidation peptide.Fermentation process, can be by microorganism producing enzyme and enzyme hydrolysis compared with enzymatic isolation method
Two steps are unified, and eliminate separation and the purification step of enzyme, reduce production process, reduce cost；In addition, microorganism is except production
Outside raw albumen enzyme, moreover it is possible to produce cellulase, phytase etc., cell membrane can be decomposed, is conducive to the activity of albumen and protease
Site is combined, and improves hydrolysis efficiency, and the peptide ending enzyme that the antioxidation activity for increasing enzymolysis product and microorganism produce is to small peptide end
Modification, make obtained peptide that there is the natural aromatic taste of fermentation, palatability is more preferable.
The fermentation method of existing report prepares anti-oxidation peptide, is concentrated mainly on peanut, rapeseed dregs, dregs of beans, Fish protein, whey egg
White to wait raw material, its effect is unsatisfactory, and has no the relevant report that anti-oxidation peptide is prepared using microbial fermentation soft-shelled turtle egg.
The content of the invention
It is contemplated that overcoming the deficiencies in the prior art to prepare anti-oxidation peptide liquid there is provided a kind of step fermentation soft-shelled turtle egg
Method.
In order to achieve the above object, the technical scheme that provides of the present invention is：
The method that the step fermentation soft-shelled turtle egg prepares anti-oxidation peptide liquid comprises the following steps：
（1）Soft-shelled turtle egg is cleaned and dried, and illumination is carried out to cleaning and dried soft-shelled turtle egg with refractor and is selected, is removed
Rotten soft-shelled turtle egg is removed, the soft-shelled turtle egg of health is obtained；
（2）The soft-shelled turtle egg chosen is put into mixer, the aseptic deionized water of 2~5 times of soft-shelled turtle egg volumes is injected, mixing time is
5~25min, 200~800r/min of mixing speed；
（3）Disc centrifuge is centrifuged, 3000~6000rpm of rotating speed, 10~20min of time, removes a small amount of grease on upper strata
With the soft-shelled turtle eggshell of bottom, soft-shelled turtle egg film, egg white, the yolk mixed liquor in middle level are obtained（That is soft-shelled turtle egg mixed liquor）；
（4）Soft-shelled turtle egg mixed liquor is added into 10 ~ 20g of glucose/L, under the conditions of 100~120 DEG C, 0.08~0.1Mpa of pressure
Sterilize 15~30min；
（5）Soft-shelled turtle egg mixed liquor after sterilizing is cooled to after room temperature, Bacillus subtilis strain seed culture fluid is added, carried out
Liquid fermentation, fermentation temperature is 32~38 DEG C, and fermentation initial pH value taste 7.0~7.5, fermentation time is 12~36h, the withered grass
Bacillus seed culture fluid kind amount accounts for the 3~8% of soft-shelled turtle egg mixeding liquid volume；
（6）To through step（5）Lactic acid bacteria culturers seed culture fluid is added in soft-shelled turtle egg mixed liquor after liquid fermentation processing, is carried out
The static fermentation of liquid, fermentation temperature is 37~47 DEG C, and fermentation initial pH value taste 6.0~6.5, fermentation time is 12~24h, described
Lactic acid bacteria culturers seed culture fluid kind amount accounts for the 3~8% of soft-shelled turtle egg mixeding liquid volume；
（7）Will be through step（6）Soft-shelled turtle egg mixed liquor after the static fermentation process of liquid is warming up to 70~98 DEG C, maintain 10~
20min, inactivates strain；Room temperature is cooled to, anti-oxidation peptide liquid, filling preservation is produced.
Preferably, in step（5）Be stirred continuously during liquid fermentation, oxygen supply.
Preferably, step（5）The preparation method of the Bacillus subtilis strain seed culture fluid is as follows：By what is activated
In seed culture medium after the access sterilizing of bacillus subtilis slant strains single bacterium colony, in 180~200r/min of rotary shaker, 32
~38 DEG C of 20~24h of constant-temperature shaking culture；The seed culture medium component is：Peptone 30g/L, NaCl 5g/L, beef extract
5g/L and glucose 1g/L, pH 7.2, the also addition agar 2.5g/L if seed culture medium is solid medium.Seed culture
Based on the 20min, cooling of being sterilized at 121 DEG C.
Preferably, step（6）The preparation method of the lactic acid bacteria culturers seed culture fluid is as follows：By lactic acid bacteria slant strains
MRS fluid nutrient mediums, 37 DEG C of h of static gas wave refrigerator 36 are accessed after activation, continuous passage 5 times；The seed culture medium component
For：The g/L of beef extract 10, the g/L of peptone 10, the g/L of dusty yeast 5, the g/L of glucose 20, the g/L of sodium acetate 5, citric acid
The g/L of two ammonium 2, the g/L of Tween 80 0.1, the g/L of magnesium sulfate 0.58, g/L, p H 6.2~6.4 of manganese sulfate 0.28.121
DEG C sterilizing 15 min, cooling.
The present invention by soft-shelled turtle egg carry out liquid agitation it is broken while, remain egg shell membrane that common scrambled egg mode abandons this
One good anti-oxidation peptide raw material, obtained soft-shelled turtle egg mixed liquor, is fermented, more with bacillus subtilis and lactic acid bacteria step by step
Each zymogenic advantage has been played well, and the anti-oxidation peptide fixture of preparation has more preferable inoxidizability.
With " a kind of that soft-shelled turtle egg is carried out into the method that enzymatic isolation method hydrolysis prepares antioxidation activity liquid " of Yang Mingyi et al. application
Compare, the present invention is more suitable for industrialized production, and degree of hydrolysis is higher（It is high by 6.68%）, the obtained oxidation resistance of enzymolysis liquid more
By force（TAC is 3.46 times of former scheme, and Scavenging action to hydroxyl free radical and superoxide anion clearance rate are respectively than former scheme
High 16.27% and 8.98%）, can be applied to the fields such as cosmetics, health products, food processing.
Brief description of the drawings
Fig. 1 is scheme degree of hydrolysis contrast before and after improving；Wherein：Sample 1 is enzymolysis liquid obtained by case in front of improving（Without molecule
Membrane filtration）, sample 2 is present invention gained anti-oxidation peptide liquid；
Fig. 2 is scheme TAC contrast before and after improving；Wherein：Sample 1 is enzymolysis liquid obtained by case in front of improving（Without dividing
Sub- membrane filtration）, sample 2 is present invention gained anti-oxidation peptide liquid；
Fig. 3 is scheme Hydroxyl radical-scavenging ability contrast before and after improving；Wherein：Sample 1 is enzymolysis liquid obtained by case in front of improving（Not
Through molecule membrane filtration）, sample 2 is present invention gained anti-oxidation peptide liquid；
Fig. 4 is scheme superoxide anion Scavenging activity contrast before and after improving；Wherein：Sample 1 is enzymolysis liquid obtained by case in front of improving
（Without molecule membrane filtration）, sample 2 is present invention gained anti-oxidation peptide liquid.
Embodiment
The method that the step fermentation soft-shelled turtle egg prepares anti-oxidation peptide liquid comprises the following steps：
（1）Soft-shelled turtle egg is cleaned and dried, and illumination is carried out to cleaning and dried soft-shelled turtle egg with refractor and is selected, is removed
Rotten soft-shelled turtle egg is removed, the soft-shelled turtle egg of health is obtained；
（2）The soft-shelled turtle egg chosen is put into mixer, the aseptic deionized water of 2~5 times of soft-shelled turtle egg volumes is injected, mixing time is
5~25min, 200~800r/min of mixing speed；
（3）Disc centrifuge is centrifuged, 3000~6000rpm of rotating speed, 10~20min of time, removes a small amount of grease on upper strata
With the soft-shelled turtle eggshell of bottom, soft-shelled turtle egg film, egg white, the yolk mixed liquor in middle level are obtained（That is soft-shelled turtle egg mixed liquor）；
（4）Soft-shelled turtle egg mixed liquor is added into 10 ~ 20g of glucose/L, under the conditions of 100~120 DEG C, 0.08~0.1Mpa of pressure
Sterilize 15~30min；
（5）Soft-shelled turtle egg mixed liquor after sterilizing is cooled to after room temperature, Bacillus subtilis strain seed culture fluid is added, carried out
Liquid fermentation, fermentation temperature is 32~38 DEG C, and fermentation initial pH value taste 7.0~7.5, fermentation time is 12~36h, the withered grass
Bacillus seed culture fluid kind amount accounts for the 3~8% of soft-shelled turtle egg mixeding liquid volume；It is stirred continuously, supplies during liquid fermentation
Oxygen；
（6）To through step（5）Lactic acid bacteria culturers seed culture fluid is added in soft-shelled turtle egg mixed liquor after liquid fermentation processing, is carried out
The static fermentation of liquid, fermentation temperature is 37~47 DEG C, and fermentation initial pH value taste 6.0~6.5, fermentation time is 12~24h, described
Lactic acid bacteria culturers seed culture fluid kind amount accounts for the 3~8% of soft-shelled turtle egg mixeding liquid volume；
（7）Will be through step（6）Soft-shelled turtle egg mixed liquor after the static fermentation process of liquid is warming up to 70~98 DEG C, maintain 10~
20min, inactivates strain；Room temperature is cooled to, anti-oxidation peptide liquid, filling preservation is produced.
Wherein, step（5）The preparation method of the Bacillus subtilis strain seed culture fluid is as follows：It is withered by what is activated
In seed culture medium after the access sterilizing of careless bacillus slant strains single bacterium colony, in 180~200r/min of rotary shaker, 32~
38 DEG C of 20~24h of constant-temperature shaking culture；The seed culture medium component is：Peptone 30g/L, NaCl 5g/L, beef extract
5g/L and glucose 1g/L, pH 7.2, the also addition agar 2.5g/L if seed culture medium is solid medium.Seed culture
Based on the 20min, cooling of being sterilized at 121 DEG C.Step（6）The preparation method of the lactic acid bacteria culturers seed culture fluid is as follows：
Lactic acid bacteria slant strains are activated, MRS fluid nutrient mediums, 37 DEG C of h of static gas wave refrigerator 36 are accessed after continuous passage 5 times；Institute
Stating seed culture medium component is：The g/L of beef extract 10, the g/L of peptone 10, the g/L of dusty yeast 5, the g/L of glucose 20, vinegar
The sour g/L of sodium 5, the g/L of dibasic ammonium citrate 2, the g/L of Tween 80 0.1, the g/L of magnesium sulfate 0.58, manganese sulfate 0.28 g/L, p
H 6.2~6.4.121 DEG C of 15 min of sterilizing, cooling.
As a result determine：
1st, relative hydrolysis degree（DH）Determine：Trichloroacetic acid method
Trichloroacetic acid（TCA）For strong acid, acted on good albumen precipitation, so as to remove the protein of macromolecular.Take enzymolysis
Liquid 10ml, adds 10%TCA solution, shakes up and stand 30min, 10000g, centrifuges 15min, makes unhydrolysed macro-molecular protein
Precipitation, is separated with small soluble molecules albumen, is determined the content that soluble protein accounts for total protein, is tried to achieve degree of hydrolysis, you can dissolubility
Albumen accounts for the percentage of total protein.
Degree of hydrolysis calculation formula：
DH（%）=（N1- N0）/ N*100%
In formula：Soluble Protein Contents (mg) before N0--- proteolysis；
Soluble Protein Contents (mg) after N1--- proteolysis；
N--- total protein contents (mg, Coomassie brilliant G-250 dye method).
Each sample repeats 3 groups, the anti-oxidation peptide liquid obtained for final degree of hydrolysis, fermentation method of averaging（Sample 2）Water
Xie Du is 20.46%, compares protease hydrolytic（That is, it is " a kind of that soft-shelled turtle egg is carried out into enzymatic isolation method hydrolysis preparation that Yang Mingyi et al. applies
The method of antioxidation activity liquid " CN201510150779.X）Obtained enzymolysis liquid（Without molecule membrane filtration）（Sample 1, hydrolysis
Spend for 13.78%）6.68% is higher by, Fig. 1 is referred to.
2nd, inoxidizability ability is determined（Use Beijing Suo Laibao Science and Technology Ltd TAC（T-AOC）Detection
Kit（FRAP methods）AAS BC1310）
Various polyphenoils and antioxidase etc. constitute total antioxidation level in measure object.
Principle：Under sour environment, Fe is reduced3+Azine (the Fe of-three pyridine three3+- TPTZ) produce blue Fe2+- TPTZ ability
Reflect TAC.
TAC is calculated：
TAC（Unit/milligram）= (1÷0.6308)×（A△-0.1291）Before ÷ sample qualities × sample measures
Extension rate.
Respectively manage net absorbance change value A △=OD △ and determine pipe-OD control tubes, be i.e. A △ correspond to standard curve
OD593。
Each sample repeats 3 groups, the anti-oxidation peptide liquid obtained for final TAC, fermentation method of averaging（Sample
2）Total oxidability be 190.53 units/grams, the enzymolysis liquid that protease hydrolytic is obtained（Without molecule membrane filtration）（Sample 1, always
Oxidability is 55.10 units/grams）3.46 times, refer to Fig. 2.
3rd, Hydroxyl radical-scavenging ability is determined（Determined using Beijing Suo Laibao Science and Technology Ltd Hydroxyl radical-scavenging ability
Kit AAS BC1320）
Hydroxy radical acts on the biomolecule such as vivo protein, nucleic acid, lipid, causes eucaryotic cell structure and function impaired, and then
Internal metabolic disorder is caused to cause disease.Hydroxyl radical-scavenging ability is one of important indicator of sample oxidation resistance.
Principle：H2O2/ Fe2+Hydroxy radical is produced by Fenton reactions, by Phen-Fe2+Fe in the aqueous solution2+
It is oxidized to Fe3+, cause 536nm absorbances to decline, sample reflects sample to the inhibition level of 536nm absorbance fall off rates
The ability of product scavenging hydroxyl.
Calculation formula：
Scavenging action to hydroxyl free radical D%=（- A pairs of A surveys）÷（- A pairs of A skies）×100%
A is empty, A is surveyed to, A：Blank tube, control tube and the light absorption value for determining pipe.
Each sample repeats 3 groups, the anti-oxidation peptide liquid obtained for final Hydroxyl radical-scavenging ability, fermentation method of averaging
（Sample 2）Scavenging action to hydroxyl free radical, the enzymolysis liquid obtained than protease hydrolytic（Without molecule membrane filtration）（Sample 1, hydroxyl is free
Base clearance rate is 30.46%）It is high by 16.27%, refer to Fig. 3.
4th, superoxide anion Scavenging activity is determined（Energy is removed using Beijing Suo Laibao Science and Technology Ltd superoxide anion
Power kit AAS BC1410）
Ultra-oxygen anion free radical can attack large biological molecule, such as a kind of free radical produced in organism metabolic process
Lipid, protein, nucleic acid and polyunsaturated fatty acids etc., make it interlink or be broken, cause the destruction of eucaryotic cell structure and function,
There is very close relationship with body aging and lesion, the research for removing ultra-oxygen anion free radical has been obtained for extensive pass
Note.
Measuring principle
AP-TEMED systems produce superoxide anion, with hydroxylamine hydrochloride reaction generation NO2 -, NO2 -With p-aminobenzene sulfonic acid and α-
The azo-compound of the effect generation red of naphthylamines, there is characteristic absorption peak, removing energy of the sample to superoxide anion at 530nm
Power and 530nm light absorption value are negatively correlated.
Calculation formula：
Superoxide anion clearance rate I%=（A control tubes-A determines pipe）/ A control tubes * 100%
Each sample repeats 3 groups, the anti-oxidation peptide liquid obtained for final superoxide anion Scavenging activity, fermentation method of averaging（Sample
Product 2）Superoxide anion clearance rate be 35.18%, the enzymolysis liquid obtained than protease hydrolytic（Without molecule membrane filtration）（Sample
1, superoxide anion clearance rate is 26.20%）It is high by 8.98%, refer to Fig. 4.
Claims (4)
1. a kind of method that step fermentation soft-shelled turtle egg prepares anti-oxidation peptide liquid, it is characterised in that methods described comprises the following steps：
（1）Soft-shelled turtle egg is cleaned and dried, and illumination is carried out to cleaning and dried soft-shelled turtle egg with refractor and is selected, is removed
Rotten soft-shelled turtle egg is removed, the soft-shelled turtle egg of health is obtained；
（2）The soft-shelled turtle egg chosen is put into mixer, the aseptic deionized water of 2~5 times of soft-shelled turtle egg volumes is injected, mixing time is
5~25min, 200~800r/min of mixing speed；
（3）Disc centrifuge is centrifuged, 3000~6000rpm of rotating speed, 10~20min of time, removes a small amount of grease on upper strata
With the soft-shelled turtle eggshell of bottom, soft-shelled turtle egg film, egg white, the yolk mixed liquor in middle level, i.e. soft-shelled turtle egg mixed liquor are obtained；
（4）Soft-shelled turtle egg mixed liquor is added into 10 ~ 20g of glucose/L, under the conditions of 100~120 DEG C, 0.08~0.1Mpa of pressure
Sterilize 15~30min；
（5）Soft-shelled turtle egg mixed liquor after sterilizing is cooled to after room temperature, Bacillus subtilis strain seed culture fluid is added, carried out
Liquid fermentation, fermentation temperature is 32~38 DEG C, and fermentation initial pH value taste 7.0~7.5, fermentation time is 12~36h, the withered grass
Bacillus seed culture fluid kind amount accounts for the 3~8% of soft-shelled turtle egg mixeding liquid volume；
（6）To through step（5）Lactic acid bacteria culturers seed culture fluid is added in soft-shelled turtle egg mixed liquor after liquid fermentation processing, is carried out
The static fermentation of liquid, fermentation temperature is 37~47 DEG C, and fermentation initial pH value taste 6.0~6.5, fermentation time is 12~24h, described
Lactic acid bacteria culturers seed culture fluid kind amount accounts for the 3~8% of soft-shelled turtle egg mixeding liquid volume；
（7）Will be through step（6）Soft-shelled turtle egg mixed liquor after the static fermentation process of liquid is warming up to 70~98 DEG C, maintain 10~
20min, inactivates strain；Room temperature is cooled to, anti-oxidation peptide liquid, filling preservation is produced.
2. the method as described in claim 1, it is characterised in that in step（5）Be stirred continuously during liquid fermentation, oxygen supply.
3. the method as described in claim 1, it is characterised in that step（5）The Bacillus subtilis strain seed culture fluid
Preparation method it is as follows：The bacillus subtilis slant strains single bacterium colony activated is accessed in the seed culture medium after sterilizing,
In 180~200r/min of rotary shaker, 32~38 DEG C of 20~24h of constant-temperature shaking culture；The seed culture medium component is：Albumen
Peptone 30g/L, NaCl 5g/L, beef extract 5g/L and glucose 1g/L, pH 7.2, if seed culture medium is solid medium
Also addition agar 2.5g/L.
4. the method as described in claim 1, it is characterised in that step（6）The preparation of the lactic acid bacteria culturers seed culture fluid
Method is as follows：Lactic acid bacteria slant strains are activated, MRS fluid nutrient mediums, 37 DEG C of static trainings are accessed after continuous passage 5 times
Support 36 h；The seed culture medium component is：The g/L of beef extract 10, the g/L of peptone 10, the g/L of dusty yeast 5, glucose
20 g/L, the g/L of sodium acetate 5, the g/L of dibasic ammonium citrate 2, the g/L of Tween 80 0.1, the g/L of magnesium sulfate 0.58, manganese sulfate
0.28 g/L, p H 6.2~6.4.
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| CN111329071B (en) * | 2020-03-13 | 2021-08-13 | 江西康宝医药生物科技有限公司 | Preparation process of brain polypeptide |
| CN113999884A (en) * | 2021-11-05 | 2022-02-01 | 南京康瑞生物科技有限公司 | Preparation method of turtle bioactive peptide |
| CN113999884B (en) * | 2021-11-05 | 2023-09-12 | 南京康瑞生物科技有限公司 | Preparation method of turtle bioactive peptide |
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Application publication date: 20170919 |