CN108949873A - The method of polypeptide active substance is extracted from konjaku - Google Patents
The method of polypeptide active substance is extracted from konjaku Download PDFInfo
- Publication number
- CN108949873A CN108949873A CN201810352896.8A CN201810352896A CN108949873A CN 108949873 A CN108949873 A CN 108949873A CN 201810352896 A CN201810352896 A CN 201810352896A CN 108949873 A CN108949873 A CN 108949873A
- Authority
- CN
- China
- Prior art keywords
- konjaku
- active substance
- polypeptide
- polypeptide active
- extracting
- 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
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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/011—Hydrolysed proteins; Derivatives thereof from plants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Diabetes (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Botany (AREA)
- Endocrinology (AREA)
- Emergency Medicine (AREA)
- Water Supply & Treatment (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses the methods that polypeptide active substance is extracted from konjaku, specifically: fry starch of konjak is subjected to alkali carries using the NaOH solution containing useful ephedrine and obtains konjaku protein extract, then protease hydrolyzed is added to konjaku protein extract and obtains enzymolysis liquid, enzymolysis liquid isolates and purifies up to konjaku polypeptide active substance.Polypeptide active substance obtained can be made into various preparations, be applied to treatment diabetes and its complication.It has the beneficial effect that preparation method simple possible of the present invention, at low cost, process stabilizing, can remove that a large amount of salt, yield are high, obtained product purity and quality is high.
Description
Technical field
The present invention relates to bioactive ingredients extractive technique field, more particularly, to extracting polypeptide active substance from konjaku
Method.
Background technique
Konjaku (Amorphophallus rivieri) it is a kind of perennial herb tuberous plant, taro is ground for Araeceae
Belong to, monoecism, likes warm, wet and half concealment ecological environment, the whole world shares 163 kinds.China shares 21 evil spirits
Taro kind is grown in the ground such as Sichuan, Shaanxi, Ningxia, Jiangsu, Guizhou, Hunan, Yunnan mostly.The ingredient of konjaku is mainly derived from
Stem tuber.In its stem tuber contain Glucomannan and starch 70% or so, containing protein and amino acid 5% ~ 15% or so, contain there are many
Minerals 4% ~ 6% or so, also alkaloid and other compositions containing 1% ~ 2%.Konjaku has extensive edible value, medicinal valence
Value, industrial value and the application value in other each fields.Under these conditions, the research hotspot of konjaku from now on will tend to
In health food, drug exploitation and its medically application and konjaku mutually tie with agricultural, textile industry, industrial technology
High-tech product research and development of conjunction etc..Therefore, imperative to the developmental research of konjaku.
Fry starch of konjak is the by-product occurred in konjaku powder process, it is a kind of light weight and tiny dust-like
Substance is not developed and used effectively because it is with certain bad smell, waste material is largely taken as directly to abandon or treat as
Auxiliary material is added in animal feed and uses.Enhancing with people to the generally understanding and self health consciousness of konjaku edible value,
The demand of konjaku rises, and the processing enterprise of konjaku has obtained quick development, but a large amount of processing waste material also generates therewith.And
Fry starch of konjak as waste material is never effectively developed and used, and so causes the pollution of environment for a long time.Therefore, such as
Fruit rationally can not only can improve environment using fry starch of konjak, moreover it is possible to which efficent use of resources is turned waste into wealth.At present to fry starch of konjak
Research mainly has biologically active peptide, active constituent, starch and its characteristic, anti-nutritional factors, peculiar smell removal, processing waste water, fermentation
The brew etc. of cream and polypeptide wine.And the active peptide research carried out using fry starch of konjak mainly has high F value oligopeptide, ACE to inhibit
Peptide, branched-chain amino acid oligopeptides, mannatide and antioxidation polypeptide etc..Currently, extracting the side of konjaku polypeptide using fry starch of konjak
Method has acid-hydrolysis method, alkali hydrolysis method and simple step enzyme solution.But in alkaline hydrolysis process, most amino acid are not by
With the destruction of degree, and racemization phenomenon is generated, so not using this method substantially in production.Sour water solution is usually that hydrochloric acid is used to exist
It is hydrolyzed under conditions of boiling, then cools down hydrolysate.Its advantage is that hydrolysis efficiency is high, does not cause the racemization of amino acid, obtain
To be l-amino acid, but partial amino-acid can be destroyed completely, and since hydrochloric acid corrosion resistance is strong, it is desirable that hydrolysis device is necessary
There is very strong corrosion resistance, so the application of acid-hydrolysis method in production is limited by very large.Item needed for enzyme hydrolysis
Part is then relatively mild, does not generate racemization, does not also destroy amino acid, and hydrolysis time is short, coloured product is shallow, is easy to control hydrolysis
Degree.But single enzyme system is relatively mostly used during preparing konjaku polypeptide using enzymatic hydrolysis at present, and there is the hydrolysis selected
The phenomenon that enzyme is unreasonable, and konjaku proteolysis is caused to be not thorough, and the production cycle seriously extends.
The prior art such as Authorization Notice No. is the Chinese invention patent of 103436582 B of CN, discloses a kind of konjaku polypeptide
Extract, preparation method and its usage, using complex enzyme zymohydrolysis fry starch of konjak protein extract, the konjaku polypeptide finally obtained is mentioned
The content of konjaku polypeptide in object is taken to be up to 85~97%, and in addition the molecular weight of konjaku polypeptide is also provided in 350~3500Da
Konjaku polypeptide extract is in preparation treatment diabetes and the purposes of diabetic complication, and contains konjaku polypeptide extract
Hypoglycemic medicine composition, in short, the konjaku polypeptide extract and its pharmaceutical composition have medical prospect well.But the system
The bioactivity for the konjaku polypeptide that Preparation Method makes is lower, and in konjaku polypeptide extract containing 0.7% ammonium salt, to human body
There are some potential safety problemss.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation method simple possible, at low cost, process stabilizing, a large amount of salt can be removed
Divide, yield height, the high method that polypeptide active substance is extracted from konjaku of obtained product purity and quality.
The present invention in view of the above technology in the problem of mentioning, the technical solution taken are as follows:
Fry starch of konjak alkali carries are obtained konjaku protein extract, then to evil spirit by the method that polypeptide active substance is extracted from konjaku
Taro protein extract is added protease hydrolyzed and obtains enzymolysis liquid, and enzymolysis liquid isolates and purifies up to konjaku polypeptide active substance, the alkali
Contain ephedrine in extraction NaOH solution.The preparation method is flown using the waste konjaku of common konjaku powder process
The konjaku polypeptide extract of great drug value has been prepared in powder, has highly important production meaning and economic valence
Value, the preparation method simple possible, process stabilizing, reproducibility is high, and used equipment is mature equipment, and cost of investment is lower,
The polypeptide active substance rate of recovery is high.
The method that polypeptide active substance is extracted from konjaku, the specific steps are that:
Alkali carries take: being 1:30-36(g/mL by solid-liquid ratio) NaOH solution that concentration is 1.4-1.6%, alkali carries are added in fry starch of konjak
1-2h is taken, is then centrifuged for, supernatant is konjaku protein extract, and alkali can destroy the secondary key especially hydrogen of protein molecular
Key dissociates some polar groups, so that protein surface molecule charge having the same is to have solubilized work to protein
With having the characteristics that recovery rate is high, at low cost, moreover it is possible to remove the ingredients such as water-soluble sugar, starch, cellulose, obtained product
Color quality is relatively good, is suitable for industrialized production；
Protease hydrolyzed: adjusting its pH to 6.0-7.0 after being preheated to 60-70 DEG C to konjaku protein extract, pawpaw egg is added
White enzyme carries out constant temperature and digests 2-3h, its pH to 8.0-9.0 is adjusted after being cooled to 30-40 DEG C, and elastoser is added and carries out constant temperature
1.5-2.5h is digested, after hydrolysis, hydrolyzate is stood and cooled down by 95-100 DEG C of water-bath enzyme deactivation 8-15min, 6000-10000r/
20-35min is centrifuged under min, supernatant is enzymolysis liquid, and different protease act on the difference of the site on protein peptide chain, is made
Different with the composition of polypeptide at hydrolysis degree, the present invention is successively digested with different protease, can make konjaku albumen
Sufficiently enzymatic hydrolysis improves the content of polypeptide active substance in enzymolysis liquid, while the step mild, reaction easy to control with enzymatic hydrolysis condition
Process, it is at low cost the advantages that, in enzymolysis process, have no toxic side effect generation；
It isolates and purifies: enzymolysis liquid is successively separated with the ultrafiltration membrane of 6-10kDa, 3-5kDa, 0.3-1kDa, after collecting ultrafiltration
Component, after passing it through rotary evaporation concentration, be freeze-dried with vacuum freeze drier and be extracted up to konjaku polypeptide
Object, which separated by the microcellular structure of film surface to substance using the pressure difference of film two sides, when the mixing of liquid
Under a certain pressure, small molecule solute penetrates film by the micropore on film to object, and macromolecular is then trapped, to realize size substance
Separation, concentration and purification, have cheap, easy to operate, separation process do not undergo phase transition, be nontoxic and pollution-free, can be effective
Protect the advantages that activity by isolated polypeptide in ground.
Preferably, containing the ephedrine of 0.32-0.43mM in NaOH solution in alkali extraction step, it is left in the ephedrine
The ratio for revolving body and d-isomer is 100:2.2-2.7.The pH that the addition of above-mentioned ephedrine is able to maintain winged powder liquid is constant, so that alkali
The rate for extracting konjaku albumen is constant, can be avoided microbial contamination caused by due to overlong time while energy saving, together
When can reduce the dosage of NaOH solution, avoid the purity for improving product in final polypeptide active substance there are a large amount of salt and
Quality, and can be avoided the waste of soda acid, reduce extraction cost；And the reasonable ratio of levo form and d-isomer can be improved protein
The binding force of molecule and amino, carboxyl, can the peptide bond of protected protein matter do not destroyed by NaOH, prevent the space of protein molecule from tying
Structure is destroyed and causes protein denaturation, improves the solubility and yield of protein, the final yield for improving polypeptide active substance.
Preferably, ultrafiltration pressure is 0.033-0.06MPa in purification procedures, the ultrafiltration time is 15-25min.
Preferably, polypeptide active substance can be made into various preparations, it is applied to treatment diabetes and its complication.The polypeptide
The blood glucose, glycosylated hemoglobin and Microalbuminuria of STZ induced diabetic rats can be significantly reduced in active material, right
Diabetes have significant therapeutic effect, and are able to achieve the stabilization hypoglycemic effect of diabetic, are greatly improved diabetic's
The adverse reaction that chemical synthetic drug generates human body can also be reduced or avoided in compliance after, and it is strong to meet human body
Health needs.
Compared with the prior art, the advantages of the present invention are as follows:
1) preparation method of the present invention utilizes the waste fry starch of konjak of common konjaku powder process, and weight has been prepared
The konjaku polypeptide extract of big drug value, has highly important production meaning and economic value, and the preparation method is simple
Viable process is stablized, and reproducibility is high, and used equipment is mature equipment, and cost of investment is lower, the polypeptide active substance rate of recovery
It is high；2) preparation method of the present invention is avoided that in final polypeptide active substance the purity and quality that product is improved there are a large amount of salt,
Extraction cost is reduced, the solubility and yield of protein are improved, the final yield for improving polypeptide active substance；3) present invention is made
Polypeptide active substance the blood glucose, glycosylated hemoglobin and microdose urine protein of STZ induced diabetic rats can be significantly reduced
Level has significant therapeutic effect to diabetes, is able to achieve the stabilization hypoglycemic effect of diabetic, is greatly improved diabetes
The adverse reaction that chemical synthetic drug generates human body can also be reduced or avoided in the compliance of patient after, meet
Human health needs.
Specific embodiment
The present invention program is described further below by embodiment:
Embodiment 1:
Fry starch of konjak alkali carries are obtained konjaku protein extract, then to evil spirit by the method that polypeptide active substance is extracted from konjaku
Taro protein extract is added protease hydrolyzed and obtains enzymolysis liquid, and enzymolysis liquid isolates and purifies up to konjaku polypeptide active substance, the alkali
Contain ephedrine in extraction NaOH solution.The preparation method is flown using the waste konjaku of common konjaku powder process
The konjaku polypeptide extract of great drug value has been prepared in powder, has highly important production meaning and economic valence
Value, the preparation method simple possible, process stabilizing, reproducibility is high, and used equipment is mature equipment, and cost of investment is lower,
The polypeptide active substance rate of recovery is high.
The method that polypeptide active substance is extracted from konjaku, the specific steps are that:
1) alkali carries take: being 1:30(g/mL by solid-liquid ratio) NaOH solution that concentration is 1.6% is added in fry starch of konjak, alkali carries take 1h,
It is then centrifuged for, supernatant is konjaku protein extract, and alkali can destroy the secondary key especially hydrogen bond of protein molecular, make
Polar group dissociates, so that protein surface molecule charge having the same to have solubilization to protein, has and mentions
Take rate height, feature at low cost, moreover it is possible to remove the ingredients such as water-soluble sugar, starch, cellulose, obtained product color quality ratio
Preferably, it is suitable for industrialized production；
2) protease hydrolyzed: adjusting its pH to 6.0 after being preheated to 70 DEG C to konjaku protein extract, be added papain into
Row constant temperature digests 3h, its pH to 9.0 is adjusted after being cooled to 30 DEG C, and elastoser is added and carries out constant temperature enzymatic hydrolysis 1.5h, hydrolysis knot
Hydrolyzate is stood and is cooled down by Shu Hou, 100 DEG C of water-bath enzyme deactivation 8min, and 20min is centrifuged under 10000r/min, and supernatant is to digest
Liquid, different protease act on the difference of the site on protein peptide chain, cause composition difference of the hydrolysis degree with polypeptide, this hair
It is bright successively to be digested with different protease, konjaku albumen can be made sufficiently to digest, improve polypeptide active substance in enzymolysis liquid
Content, while the step has many advantages, such as mild enzymatic hydrolysis condition, reaction process easy to control, at low cost, nontoxic in enzymolysis process
Side effect generates；
3) it isolates and purifies: enzymolysis liquid is successively separated with the ultrafiltration membrane of 10kDa, 5kDa, 1kDa, the group after collecting ultrafiltration
Point, after passing it through rotary evaporation concentration, be freeze-dried with vacuum freeze drier up to konjaku polypeptide extract,
The content of active peptides 92.2%, the step be using film two sides pressure difference by the microcellular structure of film surface to substance into
Row separation, when liquid mixture under a certain pressure, small molecule solute penetrates film by the micropore on film, and macromolecular then cut
It stays, to realize the separation of size substance, concentration and purification, there is cheap, easy to operate, separation process phase does not occur
Become, is nontoxic and pollution-free, the advantages that activity by isolated polypeptide can be effectively protected.
Containing the ephedrine of 0.32mM in NaOH solution in alkali extraction step, levo form and d-isomer in the ephedrine
Ratio is 100:2.7.The pH that the addition of above-mentioned ephedrine is able to maintain winged powder liquid is constant, so that alkali carries take the rate of konjaku albumen
It is constant, it can be avoided microbial contamination caused by due to overlong time while energy saving, while NaOH solution can be reduced
Dosage, avoid in final polypeptide active substance that there are a large amount of salt, improve the purity and quality of product, and can be avoided soda acid
Waste, reduce extraction cost；And the reasonable knot than can be improved protein molecule and amino, carboxyl of levo form and d-isomer
With joint efforts, can the peptide bond of protected protein matter do not destroyed by NaOH, prevent the space structure of protein molecule to be destroyed and cause albumen
Qualitative change improves the solubility and yield of protein, the final yield for improving polypeptide active substance.
Ultrafiltration pressure is 0.033MPa in purification procedures, the ultrafiltration time is 25min.
Polypeptide active substance can be made into various preparations, be applied to treatment diabetes and its complication.The polypeptide active substance
The blood glucose, glycosylated hemoglobin and Microalbuminuria that STZ induced diabetic rats can be significantly reduced, there is diabetes
Significant therapeutic effect is able to achieve the stabilization hypoglycemic effect of diabetic, and the medication for being greatly improved diabetic is complied with
Property, the adverse reaction that chemical synthetic drug generates human body can also be reduced or avoided after, meet human health needs.
Embodiment 2:
The method that polypeptide active substance is extracted from konjaku, the specific steps are that:
1) alkali carries take: being 1:33(g/mL by solid-liquid ratio) NaOH solution that concentration is 1.5% is added in fry starch of konjak, alkali carries take
1.5h is then centrifuged for, and supernatant is konjaku protein extract, and the ephedrine containing 0.38mM in above-mentioned NaOH solution is described
The ratio of levo form and d-isomer is 100:2.5 in ephedrine；
2) protease hydrolyzed: adjusting its pH to 6.5 after being preheated to 65 DEG C to konjaku protein extract, be added papain into
Row constant temperature digests 2.5h, its pH to 8.5 is adjusted after being cooled to 37 DEG C, and elastoser is added and carries out constant temperature enzymatic hydrolysis 2h, hydrolysis knot
Hydrolyzate is stood and is cooled down by Shu Hou, 98 DEG C of water-bath enzyme deactivation 12min, and 28min is centrifuged under 8000r/min, and supernatant is to digest
Liquid；
3) it isolates and purifies: enzymolysis liquid successively being separated with the ultrafiltration membrane of 8kDa, 4kDa, 0.5kDa, wherein ultrafiltration pressure is
0.05MPa, ultrafiltration time are 20min, the component after collecting ultrafiltration, after passing it through rotary evaporation concentration, with vacuum refrigeration
Drying machine be freeze-dried up to konjaku polypeptide extract, and the content of active peptides is 93.5%.
Embodiment 3:
The method that polypeptide active substance is extracted from konjaku, advanced optimizes step are as follows:
1) alkali carries take: being 1:33(g/mL by solid-liquid ratio) NaOH solution that concentration is 1.5% is added in fry starch of konjak, alkali carries take
1.5h is then centrifuged for, and supernatant is konjaku protein extract, and the ephedrine containing 0.38mM in above-mentioned NaOH solution is described
The ratio of levo form and d-isomer is 100:2.5 in ephedrine；
2) protease hydrolyzed: will be preheated to 65 DEG C to konjaku protein extract, adjust its pH to 6.5 with phosphate buffer, add
Enter papain and carry out constant temperature enzymatic hydrolysis 2.5h, its pH to 8.5 is adjusted after being cooled to 37 DEG C, elastoser is added and carries out constant temperature
2h is digested, after hydrolysis, hydrolyzate is stood and is cooled down by 98 DEG C of water-bath enzyme deactivation 12min, it is centrifuged 28min under 8000r/min, on
Clear liquid is enzymolysis liquid, the Huang of the nicotinamide-adenine dinucleotide phosphate containing 0.27mM and 0.2mM in phosphate buffer
One side of addition of plain adenine dinucleotide disodium salt, nicotinamide-adenine dinucleotide phosphate and flavin adenine dinucleotide (FAD)
Face is able to maintain that the balance of hydrogen ion and hydroxide ion concentration in phosphate-buffered, further increases the buffering of phosphate-buffered
Ability, and then guarantee that pH is relatively stable in enzymatic hydrolysis system, on the other hand can be avoided ingredient in phosphate buffer and calcium from
Sub- Ca2+、Mg2+And some heavy metal ion associate to form precipitating, guarantee that konjaku albumen comes into full contact with enzyme, improve enzymatic hydrolysis rate,
The influence that the buffer can offset to a certain extent, mitigate enzymolysis product to solution acid alkalinity, to keep the pH value of solution
It is relatively stable, the pH for adjusting enzymatic hydrolysis system always in enzymolysis process is not needed, the rate and enzymatic hydrolysis degree of enzymatic hydrolysis are improved；
3) it isolates and purifies: enzymolysis liquid successively being separated with the ultrafiltration membrane of 8kDa, 4kDa, 0.5kDa, wherein ultrafiltration pressure is
0.05MPa, ultrafiltration time are 20min, the component after collecting ultrafiltration, after passing it through rotary evaporation concentration, with vacuum refrigeration
Drying machine be freeze-dried up to konjaku polypeptide extract, and the content of active peptides is 94.8%.
Routine operation in operating procedure of the invention is well known to those skilled in the art, herein without repeating.
Technical solution of the present invention is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in spirit of the invention,
Supplement or similar fashion substitution etc., should all be included in the protection scope of the present invention.
Claims (7)
1. extracting the method for polypeptide active substance from konjaku, it is characterised in that: mention fry starch of konjak alkali carries acquirement konjaku albumen
Liquid is taken, protease hydrolyzed then is added to konjaku protein extract and obtains enzymolysis liquid, enzymolysis liquid isolates and purifies living up to konjaku polypeptide
Property substance, the alkali carries, which are taken, contains ephedrine in NaOH solution.
2. the method according to claim 1 for extracting polypeptide active substance from konjaku, it is characterised in that: the alkali carries take
Specific steps are as follows: by solid-liquid ratio be 1:30-36(g/mL) by fry starch of konjak be added concentration be 1.4-1.6% NaOH solution, alkali
1-2h is extracted, is then centrifuged for, supernatant is konjaku protein extract.
3. the method according to claim 1 for extracting polypeptide active substance from konjaku, it is characterised in that: the NaOH is molten
Containing the ephedrine of 0.32-0.43mM in liquid, the ratio of levo form and d-isomer is 100:2.2-2.7 in the ephedrine.
4. the method according to claim 1 for extracting polypeptide active substance from konjaku, it is characterised in that: the protease
The specific steps of enzymatic hydrolysis are as follows: adjust its pH to 6.0-7.0 after 60-70 DEG C being preheated to konjaku protein extract, pawpaw is added
Protease carries out constant temperature and digests 2-3h, its pH to 8.0-9.0 is adjusted after being cooled to 30-40 DEG C, and elastoser is added and carries out perseverance
Hydrolyzate is stood and is cooled down by temperature enzymatic hydrolysis 1.5-2.5h, enzyme deactivation after hydrolysis, and centrifugation, supernatant is enzymolysis liquid.
5. the method according to claim 1 for extracting polypeptide active substance from konjaku, it is characterised in that: the separation is pure
The specific steps of change are as follows: enzymolysis liquid is successively separated with the ultrafiltration membrane of 6-10kDa, 3-5kDa, 0.3-1kDa, ultrafiltration is collected
Component afterwards is freeze-dried up to konjaku polypeptide extract after concentration.
6. the method according to claim 5 for extracting polypeptide active substance from konjaku, it is characterised in that: the ultrafiltration pressure
Power is 0.033-0.06MPa, the ultrafiltration time is 15-25min.
7. the method according to claim 1 for extracting polypeptide active substance from konjaku, it is characterised in that: the polypeptide is living
Property substance can be made into various preparations, be applied to treatment diabetes and its complication.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810352896.8A CN108949873A (en) | 2018-04-19 | 2018-04-19 | The method of polypeptide active substance is extracted from konjaku |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810352896.8A CN108949873A (en) | 2018-04-19 | 2018-04-19 | The method of polypeptide active substance is extracted from konjaku |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108949873A true CN108949873A (en) | 2018-12-07 |
Family
ID=64499112
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810352896.8A Pending CN108949873A (en) | 2018-04-19 | 2018-04-19 | The method of polypeptide active substance is extracted from konjaku |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108949873A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111778306A (en) * | 2020-08-03 | 2020-10-16 | 无锡海思瑞科技有限公司 | Clean production method capable of industrially producing vegetable protein peptide |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103436582A (en) * | 2013-07-10 | 2013-12-11 | 马杰坤 | Konjac polypeptide extract, as well as preparation method and application thereof |
| CN105713943A (en) * | 2016-03-08 | 2016-06-29 | 山西大学 | Method for preparing hulless oat blood sugar reducing polypeptide with hulless oat albumen powder |
| CN105838767A (en) * | 2016-05-11 | 2016-08-10 | 陕西科技大学 | Method for preparing ACE inhibitory peptide through enzymolysis of konjac fly powder |
| CN106755243A (en) * | 2016-12-29 | 2017-05-31 | 陕西科技大学 | A kind of preparation method of konjaku albumen source ACE inhibitor peptides |
| CN108034684A (en) * | 2017-12-26 | 2018-05-15 | 金华市铁骑士生物科技有限公司 | The preparation method of sweet potato polypeptide |
-
2018
- 2018-04-19 CN CN201810352896.8A patent/CN108949873A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103436582A (en) * | 2013-07-10 | 2013-12-11 | 马杰坤 | Konjac polypeptide extract, as well as preparation method and application thereof |
| CN105713943A (en) * | 2016-03-08 | 2016-06-29 | 山西大学 | Method for preparing hulless oat blood sugar reducing polypeptide with hulless oat albumen powder |
| CN105838767A (en) * | 2016-05-11 | 2016-08-10 | 陕西科技大学 | Method for preparing ACE inhibitory peptide through enzymolysis of konjac fly powder |
| CN106755243A (en) * | 2016-12-29 | 2017-05-31 | 陕西科技大学 | A kind of preparation method of konjaku albumen source ACE inhibitor peptides |
| CN108034684A (en) * | 2017-12-26 | 2018-05-15 | 金华市铁骑士生物科技有限公司 | The preparation method of sweet potato polypeptide |
Non-Patent Citations (1)
| Title |
|---|
| 王乐,等: "麻黄碱检测方法研究进展", 《云南警官学院学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111778306A (en) * | 2020-08-03 | 2020-10-16 | 无锡海思瑞科技有限公司 | Clean production method capable of industrially producing vegetable protein peptide |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101228918A (en) | Method of preparing walnut polypeptide powder | |
| CN103052717A (en) | Industrial production method for producing antihypertensive bioactive peptide | |
| CN101642231B (en) | Drumstick mushroom essence and preparation method thereof | |
| CN108486202B (en) | Moringa oleifera hydrolysate and preparation method and application thereof | |
| WO2015176558A1 (en) | Composition of natural vitamin c and fish scale collagen peptide and preparation method thereof | |
| CN113249421B (en) | Golden-silk-royal chrysanthemum protein polypeptide and preparation and application thereof | |
| CN108753894A (en) | A kind of extracting method of American Ginseng peptide | |
| CN102030834A (en) | Method for extracting and preparing camellia polysaccharide from camellia and application of camellia polysaccharide | |
| CN102228125B (en) | Preparation method of algal active peptide | |
| CN106359839A (en) | Extraction method of oyster peptides | |
| CN108546281A (en) | A kind of ginseng oligopeptide and its preparation method and application | |
| CN101524167A (en) | Small-molecule sea cucumber powder and small-molecule sea cucumber powder capsules | |
| CN108949873A (en) | The method of polypeptide active substance is extracted from konjaku | |
| CN114134190A (en) | Preparation method of zein active peptide-carried calcium ion nano chelate | |
| WO2021082311A1 (en) | Pea peptide having supplementary blood glucose reducing function and preparation method therefor | |
| CN110734475B (en) | Oligopeptide with alpha-glucosidase inhibitory activity and application thereof | |
| CN107048417A (en) | A kind of preparation method of the peach kernel polypeptide oral liquor of high bioavilability | |
| CN1276723C (en) | Water soluble royal jelly and its dry powder and method for preparing them | |
| CN107691760A (en) | A kind of oyster peptide of enhancing development and its preparation method and application | |
| CN107488695A (en) | A kind of preparation method of the nostoc albumen of promotion plant growth | |
| CN114317658A (en) | Sea-buckthorn seed meal protein peptide composition and preparation method and application thereof | |
| CN111011869A (en) | Taihe silky fowl peptide tablet and preparation method thereof | |
| CN108486201B (en) | Method for extracting bioactive polypeptide of ginkgo | |
| CN1458263A (en) | Sea cucumber nutritive wine and its producing method and use | |
| CN111631266A (en) | Preparation method of peptide-containing high-protein soybean milk powder |
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 |
Application publication date: 20181207 |
|
| RJ01 | Rejection of invention patent application after publication |