CN106282287A - A kind of method extracting Semen Ginkgo biologically active polypeptide - Google Patents

A kind of method extracting Semen Ginkgo biologically active polypeptide Download PDF

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CN106282287A
CN106282287A CN201610896602.9A CN201610896602A CN106282287A CN 106282287 A CN106282287 A CN 106282287A CN 201610896602 A CN201610896602 A CN 201610896602A CN 106282287 A CN106282287 A CN 106282287A
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semen ginkgo
solution
active polypeptide
time
biologically active
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CN106282287B (en
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范文杰
孙俊松
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XUZHOU GINKGO RESOURCE BIO-ENGINEERING Co Ltd
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XUZHOU GINKGO RESOURCE BIO-ENGINEERING Co Ltd
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00Preparation of peptides or proteins
    • C12P21/06Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K1/00General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14Extraction; Separation; Purification
    • C07K1/145Extraction; Separation; Purification by extraction or solubilisation

Abstract

The invention discloses a kind of method extracting Semen Ginkgo biologically active polypeptide, comprise the following steps: pulverize: obtain Semen Ginkgo powder;Enzymolysis: by Semen Ginkgo powder water dissolution, stirring and evenly mixing regulation pH value, add wall breaking enzyme, pectase, cellulase and protease, enzymolysis while stirring, obtain enzymolysis solution, then through high temperature enzyme denaturing, obtain enzyme denaturing solution;Extract through two steps: adjusting enzyme denaturing solution is 35 DEG C, keeps 1 3h, hydrolyze, separate acquisition supernatant for the first time and first time residue;First time residue water again is carried and separates, collects second time supernatant;Concentrate: merge first and second supernatant, concentrate, it is thus achieved that concentrated solution;It is dried: concentrated solution is dried, obtains Semen Ginkgo biologically active polypeptide.The present invention extracts the method for Semen Ginkgo biologically active polypeptide, both ensure that the structure of active polypeptide was not destroyed, and is also effectively increased extraction ratio, and micromolecule active polypeptide molecular weight is many at 1000Da (dalton) below, is more beneficial for absorption of human body.

Description

A kind of method extracting Semen Ginkgo biologically active polypeptide
Technical field
The invention belongs to Semen Ginkgo product processing technique field, be specifically related to a kind of side extracting Semen Ginkgo biologically active polypeptide Method.
Background technology
Biologically active polypeptide is that research recently finds that the metabolic activity of cell function or Living organism is had optimization merit The small peptide class chemical molecules of energy.Compared with the protein of complete length, the physicochemical property that biologically active polypeptide is excellent, as good Dissolubility, hydrophilic, low viscosity, low foaming etc., the bioavailability all making active polypeptide is higher, even than aminoacid more Easily absorbing, have effect of regulation human physiological functions, it has not available for some crude protein or its component aminoacid Biological activity.The biological function of biologically active peptide includes immunomodulating, resisting hypertension, antibacterial, anticancer, antioxidation, improvement Element absorption, regulation flavour of food products, taste and hardness etc., therefore, biologically active polypeptide be widely used in food, health product and In cosmetics.
The classification of biologically active polypeptide can be divided by raw material sources and health care.According to having that raw material divides: breast Peptide, soybean peptide, corn peptide, fish and shellfish albumen, collagen protein polypeptide etc.;Divide according to function and have immune-active peptides, neural activity Peptide, blood pressure lowering peptide and antibacterial polypeptide etc..These polypeptide achieve industrialized production as health food and medicine, and obtain huge Big economic benefit.But the research of bioactive peptide is started late by China, and product category is few, market development is not enough.In the last few years Along with people's health and beauty consciousness and the raising of the consuming capacity, the polypeptide series products with good biological activity has entered into usually In the lives of the people, there is market widely and application prospect.Additionally, China's living species enriches, protein resource is sufficient, Biotechnology exploitation polypeptide active substance is utilized to be an up the important channel of industrial crops.
Maximum composition in Semen Ginkgo core is starch, is secondly protein, fat and polysaccharide, possibly together with vitamin C, core Huang Element, carotene, ginkgoic acid, bilobol, the trace element of multiple beneficial health and five carbon polysaccharide, therefore have good Medical effect and dietary function.Along with protein polypeptide physiological function and the expansion of consumption market, carry out Semen Ginkgo biological activity many The preparation of peptide is one of important channel improving Semen Ginkgo core consumption face and economic interests.
The preparation of traditional biological polypeptide can be completed by chemical method, i.e. utilizes acid or alkali to carry out at the digestion of protein Reason, the method speed is fast, low cost, but polypeptide can cause certain damage, the control of polypeptide structure are less susceptible to, not only Destroy the physiologically active of polypeptide, produce poison side-product, and the chemical contaminations such as soda acid can be produced.Therefore, it is necessary to exploitation one The method that can effectively extract Semen Ginkgo biologically active polypeptide, is applied to the research and development of functional food or medicine.
Summary of the invention
It is an object of the invention to provide a kind of method extracting Semen Ginkgo biologically active polypeptide, solve chemical leaching test to many The damage that peptide causes, the polypeptide structure caused is unstable, destroys polypeptide physiologically active, produces poison side-product, produces soda acid etc. and changes Learn the problem polluted.
A kind of method extracting Semen Ginkgo biologically active polypeptide that the present invention provides, comprises the following steps:
Step 1, pulverizes: Semen Ginkgo core is boiled in boiling water 3-5min, pulverizes and sieves after drying, it is thus achieved that Semen Ginkgo powder;
Step 2, enzymolysis: by the Semen Ginkgo powder water dissolution in step 1, wherein Semen Ginkgo powder is 1g:10-with the solid-to-liquid ratio of water 30mL, stirring and evenly mixing, regulate pH to 5.5-7.0, while stirring, add wall breaking enzyme, pectase, cellulase and protease, At 55-60 DEG C, enzymolysis 1-4h, obtains enzymolysis solution, then by enzymolysis solution through high temperature enzyme denaturing, obtains enzyme denaturing solution;
Wherein, the addition of wall breaking enzyme accounts for the 0.1-1.0% of Semen Ginkgo powder quality, and the addition of pectase accounts for Semen Ginkgo powder quality 0.1-1.0%, the addition of cellulase accounts for the 0.1-1.0% of Semen Ginkgo powder quality, and the addition of protease accounts for Semen Ginkgo opaque The 0.5-3.0% of amount;
Described wall breaking enzyme is mixed by the mass ratio of 7:1 by Snailase, lysozyme;Described protease is acidic protein One or more in enzyme, alkaline protease, neutral protease, papain, pancreatin, flavor protease, bromelain;
Step 3, two steps are extracted: adjust the temperature of enzyme denaturing solution to 3-5 DEG C, keep 1-3h, separate or centrifugal through sucking filtration Separate, it is thus achieved that hydrolyzed solution and for the first time residue for the first time;First time hydrolyzed solution is warming up to 55-60 DEG C, keeps 1-2h, then warp Centrifugation, collects supernatant for the first time;
The first time residue obtained is dissolved in the NaCl solution of 0.1-0.3mol/L or 0.01-0.1mol/L again NaH2PO4In solution, keep 1-2h in 60-80 DEG C, carry out sucking filtration separation or centrifugation the most again, collect second time supernatant, Wherein first time residue and NaCl solution or NaH2PO4The solid-to-liquid ratio of solution is 1g:10-25mL;
Step 4, concentrates: merges supernatant and second time supernatant for the first time, concentrates and remove portion of water, it is thus achieved that concentrate Liquid;
Step 5, is dried: be dried by gained concentrated solution in step 4, obtain Semen Ginkgo biologically active polypeptide.
Preferably, in step 1, the mesh number sieved is 100-2000 mesh.
Preferably, in step 2, Semen Ginkgo powder is 1g:30mL with the solid-to-liquid ratio of water.
Preferably, step 2, the method for high temperature enzyme denaturing is: in 95-100 DEG C, enzymolysis solution is incubated 5-7min.
Preferably, in step 3, when sucking filtration separates, add filter aid kieselguhr.
Preferably, in step 4, during concentration, use and be concentrated in vacuo or rotary evaporation concentration.
Preferably, in step 5, when being dried, use vacuum drying, and vacuum drying temperature is 50 DEG C.
The present invention extracts the method for Semen Ginkgo biologically active polypeptide, is processed biological sample by multiple enzyme preparation, can In a mild condition protein is separated from plant tissue, owing to not adding chemical reagent during extracting, Not only ensure that the active structure of protein is not destroyed, it is also possible to be effectively improved the extraction ratio of albumen, obtain after enzymolysis Micromolecule active polypeptide molecular weight is many at 1000Da (dalton) below, and is evenly distributed, and is more beneficial for absorption of human body.Can be used for Repairing skin, increase skin-nourishing, can increase sharply when being used in dietetic therapy proteinaceous nutrient to patient.
Compared with traditional two-step extraction, biologically active polypeptide yield is high, and the ratio of small-molecular peptides active constituent is big, nothing Disposal of pollutants.Using the Semen Ginkgo powder raw material of 1000 mesh, the yield of Enzymatic Extraction Semen Ginkgo polypeptide the most equally is more than 55%; The ratio of the little peptide active constituent of about 1000Da is more than 80%, and the ratio of the active small peptide component that traditional method obtains is not More than 60%.
Detailed description of the invention
Below the detailed description of the invention of invention is described in detail, it is to be understood that protection scope of the present invention is not subject to The restriction of detailed description of the invention.The test method of unreceipted actual conditions in the following example, generally according to normal condition, or According to the condition proposed by each manufacturer.
When embodiment provides numerical range, it should be appreciated that unless the present invention is otherwise noted, two ends of each numerical range Between point and two end points, any one numerical value all can be selected for.Unless otherwise defined, in the present invention use all technology and The same meaning that scientific terminology and those skilled in the art of the present technique are generally understood that.Except in embodiment use concrete grammar, equipment, Outside material, according to those skilled in the art's grasp to prior art and the record of the present invention, it is also possible to use and this Any method, equipment and the material of the prior art that the method described in inventive embodiments, equipment, material are similar or equivalent comes real The existing present invention.
Additionally, the various enzymes used in the present invention and the following example are commercially available.
Preferably, a kind of method extracting Semen Ginkgo biologically active polypeptide of the present invention, including following example.
Embodiment 1
A kind of method extracting Semen Ginkgo biologically active polypeptide, comprises the following steps:
Step 1, pulverizes: Semen Ginkgo core is boiled in boiling water 5min, uses crusher for Chinese herbal medicine to carry out during crushed after being dried Pulverize, cross 200 mesh sieves, it is thus achieved that the Semen Ginkgo powder of 200 mesh;
Step 2, enzymolysis: by the Semen Ginkgo powder water dissolution in step 1, wherein Semen Ginkgo powder is 1g:20mL with the solid-to-liquid ratio of water, Stirring and evenly mixing, regulates pH to 5.5, adds wall breaking enzyme, pectase, cellulase and protease, while stirring at 55 DEG C Enzymolysis 4h, obtains enzymolysis solution, then by enzymolysis solution through high temperature enzyme denaturing, obtains enzyme denaturing solution;
Wherein, the method for high temperature enzyme denaturing is: in 100 DEG C, enzymolysis solution is incubated 5min;
Wherein, described wall breaking enzyme is mixed by the mass ratio of 7:1 by Snailase, lysozyme;Described protease is by acid Property protease, neutral protease, papain, pancreatin, flavor protease composition;
Wherein, the addition of wall breaking enzyme accounts for the 0.4% of Semen Ginkgo powder quality, and the addition of pectase accounts for Semen Ginkgo powder quality 0.2%, the addition of cellulase accounts for the 0.2% of Semen Ginkgo powder quality, and the addition of acid protease accounts for Semen Ginkgo powder quality 0.5%, the addition of neutral protease accounts for the 0.2% of Semen Ginkgo powder quality, and the addition of papain accounts for Semen Ginkgo powder quality 0.5%, the addition of pancreatin accounts for the 0.3% of Semen Ginkgo powder quality, and the addition of flavor protease accounts for the 1.0% of Semen Ginkgo powder quality;
Step 3, two steps are extracted: the temperature of enzyme denaturing solution adjusted to 4 DEG C, keep 1h, are centrifuged 10min through 3500r/min, Obtain hydrolyzed solution and for the first time residue for the first time;First time hydrolyzed solution is warming up to 60 DEG C, keeps 1h, then through 12000r/min Centrifugal 10min, collects supernatant for the first time;
Again the first time residue obtained is dissolved in the NaCl solution of 0.1mol/L, in 80 DEG C keep 2h, the most again with 12000r/min is centrifuged 15min, collects second time supernatant, and wherein the solid-to-liquid ratio of residue and NaCl solution is 1g for the first time: 25mL;
Step 4, concentrates: merges supernatant and second time supernatant for the first time, utilizes Rotary Evaporators by supernatant concentration To original volume 1/10, it is thus achieved that concentrated solution;
Step 5, is dried: gained concentrated solution in step 4 carries out 50 DEG C of vacuum drying, obtains Semen Ginkgo biologically active polypeptide.
Through analyzing, in Semen Ginkgo biologically active polypeptide, molecular weight accounts for polypeptide total amount at the bioactive micro peptide of about 1000Da About 85%, total extraction ratio of biologically active polypeptide is about 60.02%.
Embodiment 2
A kind of method extracting Semen Ginkgo biologically active polypeptide, comprises the following steps:
Step 1, pulverizes: Semen Ginkgo core is boiled in boiling water 4min, uses super micron mill to carry out powder during crushed after being dried Broken, cross 500 mesh sieves, it is thus achieved that the Semen Ginkgo powder of 500 mesh;
Step 2, enzymolysis: by the Semen Ginkgo powder water dissolution in step 1, wherein Semen Ginkgo powder is 1g:25mL with the solid-to-liquid ratio of water, Stirring and evenly mixing, regulates pH to 6, adds wall breaking enzyme, pectase, cellulase and protease, enzyme at 55 DEG C while stirring Solve 3h, obtain enzymolysis solution, then by enzymolysis solution through high temperature enzyme denaturing, obtain enzyme denaturing solution;
Wherein, the method for high temperature enzyme denaturing is: in 95 DEG C, enzymolysis solution is incubated 5min;
Wherein, described wall breaking enzyme is mixed by the mass ratio of 7:1 by Snailase, lysozyme;Described protease is by acid Property protease, alkaline protease, papain, pancreatin, flavor protease, bromelain composition;
Wherein, the addition of wall breaking enzyme accounts for the 0.5% of Semen Ginkgo powder quality, and the addition of pectase accounts for Semen Ginkgo powder quality 0.5%, the addition of cellulase accounts for the 0.2% of Semen Ginkgo powder quality, and the addition of acid protease accounts for Semen Ginkgo powder quality 0.2%, the addition of alkaline protease accounts for the 1.0% of Semen Ginkgo powder quality, and the addition of papain accounts for Semen Ginkgo powder quality 0.5%, the addition of pancreatin accounts for the 1.0% of Semen Ginkgo powder quality, and the addition of flavor protease accounts for the 0.1% of Semen Ginkgo powder quality, The addition of bromelain accounts for the 0.1% of Semen Ginkgo powder quality;
Step 3, two steps are extracted: the temperature of enzyme denaturing solution adjusted to 3 DEG C, keep 3h, separate through sucking filtration, it is thus achieved that for the first time Hydrolyzed solution and for the first time residue;First time hydrolyzed solution is warming up to 60 DEG C, keeps 1h, be then centrifuged 10min through 12000r/min, Collect supernatant for the first time;
Again the first time residue obtained is dissolved in the NaCl solution of 0.3mol/L, keeps 1h in 70 DEG C, the most again through taking out Filter separates, and collects second time supernatant, and wherein first time residue is 1g:15mL with the solid-to-liquid ratio of NaCl solution;
Step 4, concentrates: merge supernatant and second time supernatant for the first time, supernatant is concentrated in vacuo to original volume 1/10, it is thus achieved that concentrated solution;
Step 5, is dried: gained concentrated solution in step 4 carries out 50 DEG C of vacuum drying, obtains Semen Ginkgo biologically active polypeptide.
Through analyzing, in Semen Ginkgo biologically active polypeptide, molecular weight accounts for polypeptide total amount at the bioactive micro peptide of about 1000Da About 82%, total extraction ratio of biologically active polypeptide is about 54.47%.
Embodiment 3
A kind of method extracting Semen Ginkgo biologically active polypeptide, comprises the following steps:
Step 1, pulverizes: Semen Ginkgo core is boiled in boiling water 3min, uses crusher for Chinese herbal medicine to carry out during crushed after being dried Pulverize, cross 1000 mesh sieves, it is thus achieved that the Semen Ginkgo powder of 1000 mesh;
Step 2, enzymolysis: by the Semen Ginkgo powder water dissolution in step 1, wherein Semen Ginkgo powder is 1g:30mL with the solid-to-liquid ratio of water, Stirring and evenly mixing regulation pH value, to 7.0, adds wall breaking enzyme, pectase, cellulase and protease, at 58 DEG C while stirring Enzymolysis 1h, obtains enzymolysis solution, then by enzymolysis solution through high temperature enzyme denaturing, obtains enzyme denaturing solution;
Wherein, the method for high temperature enzyme denaturing is: in 100 DEG C, enzymolysis solution is incubated 7min;
Wherein, described wall breaking enzyme is mixed by the mass ratio of 7:1 by Snailase, lysozyme;Described protease is by wood Melon protease, flavor protease, bromelain form;
Wherein, the addition of wall breaking enzyme accounts for the 0.1% of Semen Ginkgo powder quality, and the addition of pectase accounts for Semen Ginkgo powder quality 1.0%, the addition of cellulase accounts for the 0.8% of Semen Ginkgo powder quality, and the addition of papain accounts for Semen Ginkgo powder quality 1.0%, the addition of flavor protease accounts for the 1.0% of Semen Ginkgo powder quality, and the addition of bromelain accounts for Semen Ginkgo powder quality 1.0%;
Step 3, two steps are extracted: the temperature of enzyme denaturing solution adjusted to 5 DEG C, keep 3h, separate through sucking filtration, and during sucking filtration Add filter aid kieselguhr, it is thus achieved that hydrolyzed solution and for the first time residue for the first time;First time hydrolyzed solution is warming up to 55 DEG C, keeps 1.5h, is then centrifuged 10min through 12000r/min, collects supernatant for the first time;
The first time residue obtained is dissolved in the NaH of 0.01mol/L again2PO4In solution, keep 1.5h in 70 DEG C, the most again It is centrifuged 15min with 12000r/min, collects second time supernatant, wherein first time residue and NaH2PO4The solid-to-liquid ratio of solution is 1g:10mL;
Step 4, concentrates: merges supernatant and second time supernatant for the first time, utilizes Rotary Evaporators by supernatant concentration To original volume 1/10, it is thus achieved that concentrated solution;
Step 5, is dried: gained concentrated solution in step 4 carries out 50 DEG C of vacuum drying, obtains Semen Ginkgo biologically active polypeptide.
Through analyzing, in Semen Ginkgo biologically active polypeptide, molecular weight accounts for polypeptide total amount at the bioactive micro peptide of about 1000Da About 80%, total extraction ratio of biologically active polypeptide is about 56.62%.
Embodiment 4
A kind of method extracting Semen Ginkgo biologically active polypeptide, comprises the following steps:
Step 1, pulverizes: Semen Ginkgo core is boiled in boiling water 5min, uses crusher for Chinese herbal medicine to carry out during crushed after being dried Pulverize, cross 500 mesh sieves, it is thus achieved that the Semen Ginkgo powder of 500 mesh;
Step 2, enzymolysis: by the Semen Ginkgo powder water dissolution in step 1, wherein Semen Ginkgo powder is 1g:20mL with the solid-to-liquid ratio of water, Stirring and evenly mixing, regulates pH to 5.5, adds wall breaking enzyme, pectase, cellulase and protease, while stirring at 55 DEG C Enzymolysis 4h, obtains enzymolysis solution, then by enzymolysis solution through high temperature enzyme denaturing, obtains enzyme denaturing solution;
Wherein, the method for high temperature enzyme denaturing is: in 100 DEG C, enzymolysis solution is incubated 5min;
Wherein, described wall breaking enzyme is mixed by the mass ratio of 7:1 by Snailase, lysozyme;Described protease is by acid Property protease and neutral protease composition;
Wherein, the addition of wall breaking enzyme accounts for the 0.3% of Semen Ginkgo powder quality, and the addition of pectase accounts for Semen Ginkgo powder quality 0.2%, the addition of cellulase accounts for the 0.1% of Semen Ginkgo powder quality, and the addition of acid protease accounts for Semen Ginkgo powder quality 1.0%, the addition of neutral protease accounts for the 1.0% of Semen Ginkgo powder quality;
Step 3, two steps are extracted: the temperature of enzyme denaturing solution adjusted to 4 DEG C, keep 1h, are centrifuged 10min through 3500r/min, Obtain hydrolyzed solution and for the first time residue for the first time;First time hydrolyzed solution is warming up to 60 DEG C, keeps 1h, then through 12000r/min Centrifugal 10min, collects supernatant for the first time;
Again the first time residue obtained is dissolved in the NaCl solution of 0.1mol/L, in 80 DEG C keep 2h, the most again with 12000r/min is centrifuged 15min, collects second time supernatant, and wherein the solid-to-liquid ratio of residue and NaCl solution is 1g for the first time: 25mL;
Step 4, concentrates: merges supernatant and second time supernatant for the first time, utilizes Rotary Evaporators by supernatant concentration To original volume 1/10, it is thus achieved that concentrated solution;
Step 5, is dried: gained concentrated solution in step 4 carries out 50 DEG C of vacuum drying, obtains Semen Ginkgo biologically active polypeptide.
Through analyzing, in Semen Ginkgo biologically active polypeptide, molecular weight accounts for polypeptide total amount at the bioactive micro peptide of about 1000Da About 85%, total extraction ratio of biologically active polypeptide is about 55.13%.
Embodiment 5
A kind of method extracting Semen Ginkgo biologically active polypeptide, comprises the following steps:
Step 1, pulverizes: Semen Ginkgo core is boiled in boiling water 3min, uses crusher for Chinese herbal medicine to carry out during crushed after being dried Pulverize, cross 1000 mesh sieves, it is thus achieved that the Semen Ginkgo powder of 1000 mesh;
Step 2, enzymolysis: by the Semen Ginkgo powder water dissolution in step 1, wherein Semen Ginkgo powder is 1g:30mL with the solid-to-liquid ratio of water, Stirring and evenly mixing regulation pH value, to 7.0, adds wall breaking enzyme, pectase, cellulase and protease, at 58 DEG C while stirring Enzymolysis 1h, obtains enzymolysis solution, then by enzymolysis solution through high temperature enzyme denaturing, obtains enzyme denaturing solution;
Wherein, the method for high temperature enzyme denaturing is: in 100 DEG C, enzymolysis solution is incubated 7min;
Wherein, described wall breaking enzyme is mixed by the mass ratio of 7:1 by Snailase, lysozyme;Described protease is wood Melon protease;
Wherein, the addition of wall breaking enzyme accounts for the 0.1% of Semen Ginkgo powder quality, and the addition of pectase accounts for Semen Ginkgo powder quality 1.0%, the addition of cellulase accounts for the 0.8% of Semen Ginkgo powder quality, and the addition of papain accounts for Semen Ginkgo powder quality 1.0%;
Step 3, two steps are extracted: the temperature of enzyme denaturing solution adjusted to 5 DEG C, keep 3h, separate through sucking filtration, and during sucking filtration Add filter aid kieselguhr, it is thus achieved that hydrolyzed solution and for the first time residue for the first time;First time hydrolyzed solution is warming up to 55 DEG C, keeps 1.5h, is then centrifuged 10min through 12000r/min, collects supernatant for the first time;
The first time residue obtained is dissolved in the NaH of 0.01mol/L again2PO4In solution, keep 1.5h in 70 DEG C, the most again It is centrifuged 15min with 12000r/min, collects second time supernatant, wherein first time residue and NaH2PO4The solid-to-liquid ratio of solution is 1g:10mL;
Step 4, concentrates: merges supernatant and second time supernatant for the first time, utilizes Rotary Evaporators by supernatant concentration To original volume 1/10, it is thus achieved that concentrated solution;
Step 5, is dried: gained concentrated solution in step 4 carries out 50 DEG C of vacuum drying, obtains Semen Ginkgo biologically active polypeptide.
Through analyzing, in Semen Ginkgo biologically active polypeptide, molecular weight accounts for polypeptide total amount at the bioactive micro peptide of about 1000Da About 83%, total extraction ratio of biologically active polypeptide is about 58.32%.
The ratio of the active small peptide component (molecular weight is less than 1000Da) that traditional method obtains is less than 60%, and extraction has Efficiency is less than 45%.And from above-described embodiment acquired results, use the method for the present invention to extract Semen Ginkgo biologically active polypeptide, Molecular weight accounts for more than the 80% of polypeptide total amount at the bioactive micro peptide of about 1000Da, has raising by a relatively large margin.
Although preferred embodiments of the present invention have been described, but those skilled in the art once know basic creation Property concept, then can make other change and amendment to these embodiments.So, claims are intended to be construed to include excellent Select embodiment and fall into all changes and the amendment of the scope of the invention.
Obviously, those skilled in the art can carry out various change and the modification essence without deviating from the present invention to the present invention God and scope.So, if these amendments of the present invention and modification belong to the scope of the claims in the present invention and equivalent technologies thereof Within, then the present invention is also intended to comprise these change and modification.

Claims (7)

1. the method extracting Semen Ginkgo biologically active polypeptide, it is characterised in that comprise the following steps:
Step 1, pulverizes: Semen Ginkgo core is boiled in boiling water 3-5min, pulverizes and sieves after drying, it is thus achieved that Semen Ginkgo powder;
Step 2, enzymolysis: by the Semen Ginkgo powder water dissolution in step 1, wherein Semen Ginkgo powder is 1g:10-30mL with the solid-to-liquid ratio of water, Stirring and evenly mixing, regulates pH to 5.5-7.0, adds wall breaking enzyme, pectase, cellulase and protease, at 55-while stirring At 60 DEG C, enzymolysis 1-4h, obtains enzymolysis solution, then by enzymolysis solution through high temperature enzyme denaturing, obtains enzyme denaturing solution;
Wherein, the addition of wall breaking enzyme accounts for the 0.1-1.0% of Semen Ginkgo powder quality, and the addition of pectase accounts for Semen Ginkgo powder quality 0.1-1.0%, the addition of cellulase accounts for the 0.1-1.0% of Semen Ginkgo powder quality, and the addition of protease accounts for Semen Ginkgo powder quality 0.5-3.0%;
Described wall breaking enzyme is mixed by the mass ratio of 7:1 by Snailase, lysozyme;Described protease be acid protease, One or more in alkaline protease, neutral protease, papain, pancreatin, flavor protease, bromelain;
Step 3, two steps are extracted: adjust the temperature of enzyme denaturing solution to 3-5 DEG C, keep 1-3h, separate or centrifugation through sucking filtration, Obtain hydrolyzed solution and for the first time residue for the first time;First time hydrolyzed solution is warming up to 55-60 DEG C, keeps 1-2h, the most by centrifugation Separate, collect supernatant for the first time;
Again the first time residue obtained is dissolved in the NaCl solution of 0.1-0.3mol/L or the NaH of 0.01-0.1mol/L2PO4Molten In liquid, keep 1-2h in 60-80 DEG C, carry out sucking filtration separation or centrifugation the most again, collect second time supernatant, wherein first Secondary residue and NaCl solution or NaH2PO4The solid-to-liquid ratio of solution is 1g:10-25mL;
Step 4, concentrates: merges supernatant and second time supernatant for the first time, concentrates and remove portion of water, it is thus achieved that concentrated solution;
Step 5, is dried: be dried by gained concentrated solution in step 4, obtain Semen Ginkgo biologically active polypeptide.
The method extracting Semen Ginkgo biologically active polypeptide the most according to claim 1, it is characterised in that in step 1, sieve Mesh number is 100-2000 mesh.
The method extracting Semen Ginkgo biologically active polypeptide the most according to claim 1, it is characterised in that in step 2, Semen Ginkgo powder It is 1g:30mL with the solid-to-liquid ratio of water.
The method extracting Semen Ginkgo biologically active polypeptide the most according to claim 1, it is characterised in that in step 2, high temperature goes out The method of enzyme is: in 95-100 DEG C, enzymolysis solution is incubated 5-7min.
The method extracting Semen Ginkgo biologically active polypeptide the most according to claim 1, it is characterised in that in step 3, sucking filtration divides From time add filter aid kieselguhr.
The method extracting Semen Ginkgo biologically active polypeptide the most according to claim 1, it is characterised in that in step 4, concentrates Time, use and be concentrated in vacuo or rotary evaporation concentration.
The method extracting Semen Ginkgo biologically active polypeptide the most according to claim 1, it is characterised in that in step 5, when being dried Use vacuum drying, and vacuum drying temperature is 50 DEG C.
CN201610896602.9A 2016-10-13 2016-10-13 Method for extracting bioactive polypeptide of ginkgo Active CN106282287B (en)

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Cited By (9)

* Cited by examiner, † Cited by third party
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CN114766683A (en) * 2022-04-28 2022-07-22 威海食德源生物科技有限责任公司 Preparation method of leaf-eating grass active peptide
CN115337328A (en) * 2022-08-31 2022-11-15 山东省现代中药研究院有限公司 Preparation method of medicinal and edible traditional Chinese medicine decoction pieces based on biological enzymolysis technology

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Publication number Priority date Publication date Assignee Title
CN106834403A (en) * 2017-02-15 2017-06-13 国肽生物科技(北京)有限公司 The extracting method of ganoderma lucidum polypeptide
CN107418990B (en) * 2017-07-05 2020-12-01 南京林业大学 Ginkgo functional polypeptide and preparation method and application thereof
CN107418990A (en) * 2017-07-05 2017-12-01 南京林业大学 A kind of ginkgo functional polypeptide and its preparation method and application
CN109321621B (en) * 2017-08-01 2021-10-12 江苏天肽生物科技有限公司 Preparation method and new application of ginkgo polypeptide
CN109321621A (en) * 2017-08-01 2019-02-12 天津天肽生物科技有限公司 A kind of preparation method and its new application of gingko polypeptide
CN108148883A (en) * 2018-01-28 2018-06-12 淮阴师范学院 A kind of method that active peptides are extracted from loquat
CN108486201A (en) * 2018-04-18 2018-09-04 河南工程学院 A kind of extracting method of gingko biologically active polypeptide
CN108486201B (en) * 2018-04-18 2021-07-16 河南工程学院 Method for extracting bioactive polypeptide of ginkgo
CN108743632A (en) * 2018-04-27 2018-11-06 浙江工业大学 A kind of method of ultrasonic wave auxiliary removal Ginkgolic Acids in Fruits of Ginkgo biloba
CN111073941A (en) * 2019-09-12 2020-04-28 天津芸熙生物技术有限公司 Preparation process of sandalwood polypeptide
CN111073941B (en) * 2019-09-12 2022-10-14 天津芸熙生物技术有限公司 Preparation process of sandalwood polypeptide
CN114766683A (en) * 2022-04-28 2022-07-22 威海食德源生物科技有限责任公司 Preparation method of leaf-eating grass active peptide
CN115337328A (en) * 2022-08-31 2022-11-15 山东省现代中药研究院有限公司 Preparation method of medicinal and edible traditional Chinese medicine decoction pieces based on biological enzymolysis technology

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