CN106011207A - Method for extracting fructus cannabis oligopeptide from fructus cannabis cake - Google Patents
Method for extracting fructus cannabis oligopeptide from fructus cannabis cake Download PDFInfo
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- CN106011207A CN106011207A CN201610488576.6A CN201610488576A CN106011207A CN 106011207 A CN106011207 A CN 106011207A CN 201610488576 A CN201610488576 A CN 201610488576A CN 106011207 A CN106011207 A CN 106011207A
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- CN
- China
- Prior art keywords
- fructus cannabis
- oligopeptide
- grouts
- solution
- fructus
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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
-
- 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
Abstract
The invention provides a method for extracting fructus cannabis oligopeptide from a fructus cannabis cake. The method comprises the following steps: collecting the fructus cannabis cake with the oil content of lower than 1%, adding water, regulating the pH value to 9.0, adding sodium metabisulfite, carrying out solid-liquid separation, and collecting the liquid phase; adding a HCl solution into the liquid phase, regulating the pH value to 3.8-4.3, standing, carrying out secondary solid-liquid separation, and collecting the precipitate; adding water, heating to 40-55 DEG C, regulating the pH value to 7.0-9.0, adding a compound protease, carrying out enzymolysis for 3-6 hours, carrying out centrifugal filtration, and collecting the clear solution which is the fructus cannabis oligopeptide solution; and carrying out electrodialysis, concentration and freeze-drying to obtain the fructus cannabis oligopeptide powder. The fructus cannabis cake subjected to press deoiling is used as the raw material, the isoelectric point principle is combined to precipitate the protein, and the compound protease enzymolysis and freeze-drying are performed to obtain the fructus cannabis oligopeptide. The whole process utilizes the pure physical technique, and no chemical component is added; and thus, the method has the advantages of low cost and no environment pollution.
Description
Technical field
The present invention relates to the recovery and reuse technology field of Fructus Cannabis grouts, be specifically related to from Fructus Cannabis grouts
The method extracting Fructus Cannabis oligopeptide.
Background technology
Fructus Cannabis is the kernel of moraceae plants Fructus Cannabis (Cannabis sativa L.), has and moisturizes laxation, diuretic
Effect treating stranguria, that invigorate blood circulation, is the laxative drug that the most also medicine is also eaten.Modern study shows, Fructus Cannabis has preferably
Blood pressure lowering, paroxysmal pain, antiinflammatory, antithrombotic effect, be eaten for a long time, paralysis scorching to chronic forms,
Ulcer and gynaecopathia have good auxiliary curative effect.Fructus Cannabis grouts are to utilize Fructus Cannabis to extract Fructus Cannabis
Residue after oil, wherein protein content is up to 56%, containing the various necessary aminoacid of needed by human body, special
Not rich in arginine, lysine and glutamic acid, it is the most excellent a kind of vegetable protein new resources, Fructus Cannabis
Grouts remain Fructus Cannabis major part effect.
Containing the most oxidized aldehydes matter in Fructus Cannabis, the Fructus Cannabis cake after Fructus Cannabis processing can be remained in
In the dregs of rice, pink colour oxidation product can be produced after oxidation in atmosphere, have impact on Fructus Cannabis grouts institute converted products
Color and luster.It addition, possibly together with a part of monosaccharide in addition to containing protein in Fructus Cannabis grouts so that Urtica cannabina L.
The separating technology of core albumen is complex.
Prior art extracts egg albumen powder, and complicated process of preparation only with extraction from Fructus Cannabis,
Product purity is relatively low.And the method preparing Fructus Cannabis oligopeptide for raw material with Fructus Cannabis grouts have not been reported.
Summary of the invention
It is an object of the invention to develop a kind of Fructus Cannabis grouts to deoil through squeezing as raw material, preparation height
The method of quality Fructus Cannabis oligopeptide, it is provided that the utilization rate of Fructus Cannabis.
For reaching above-mentioned purpose, the invention provides and a kind of from Fructus Cannabis grouts, extract Fructus Cannabis oligopeptide
Method, concretely comprises the following steps:
S1, the collection fat content Fructus Cannabis grouts less than 1%, by Fructus Cannabis grouts: the mass ratio of water=1:20
In described Fructus Cannabis grouts, add water, adjust pH to 9.0 by the NaOH solution of 2.5mol/L, add described
The sodium metabisulfite of Fructus Cannabis grouts quality 1% amount, carries out solid-liquid for the first time after 70 DEG C of heated and stirred 30min
Separate, collect liquid phase；In described liquid phase, add the HCl solution of 2.5mol/L, regulate pH to 3.8～4.3
After, stand 2h and carry out second time solid-liquid separation, collect precipitation, be Fructus Cannabis separation albumen；
In Fructus Cannabis grouts lysate, add the sodium metabisulfite of 1%, can effectively prevent Fructus Cannabis cake
The oxidation of aldehydes matter in the dregs of rice.
S2, add water in the Fructus Cannabis separation albumen that step S1 prepares, described Fructus Cannabis separation albumen and water
Mass ratio is 1:15～25；It is heated to 40～55 DEG C, regulates pH to 7.0～9.0 by the NaOH solution of 2.5mol/L,
Add 1.0～5.0% compound protease of amount of described Fructus Cannabis separation albumen quality；After stirring enzymolysis 3～6h,
It is warming up to 80～95 DEG C of enzyme denaturing 10～25min, centrifugal collection clear liquid, it is Fructus Cannabis oligopeptide solution；
The quality proportioning of described compound protease is: pepsin: trypsin: flavor protease
=3～5:2～4:2～4；
S3, the Fructus Cannabis oligopeptide solution that step S2 prepares is carried out electrodialysis, is sloughed inorganic ions after, warp
Membrance concentration equipment concentrates, and obtains Fructus Cannabis oligopeptide concentrated solution；In described Fructus Cannabis oligopeptide concentrated solution solvable
Property solid content is 20～40wt%；By described Fructus Cannabis oligopeptide concentrated solution lyophilization, obtain Fructus Cannabis
Oligopeptide.
Under optimal way, solid-liquid separation described in step S1 uses automatic discharge horizontal screw centrifuge 4000rpm to be centrifuged.
Under optimal way, described in step S3, electrodialytic operating condition is: voltage 75V, enters Fructus Cannabis oligomeric
Peptide solution flow velocity 1.55 ton hour, concentrated stream speed 1.60 ton hour, cooling of electrode water flow velocity 1 ton hour.
Under optimal way, membrance concentration equipment described in step S3 uses hollow fiber nanofiltration membrane, operates pressure and is
1.5～1.6Mpa.
The inventive method has the advantage that
1, the inventive method uses the method for twice solid-liquid separation to carry out the separation of fructus cannabis, the most solid
Liquid separates and removes crude fibre, starch and fat, and solid-liquid separation removes monosaccharide for the second time；In solid-liquid separation process
In, use automatic clinker-removing horizontal screw centrifuge, improve in Fructus Cannabis grouts and imitate with Protein Separation containing part monosaccharide
Rate, makes monosaccharide mass content in albumen less than 5%, improve and separate purity of protein.
The present invention is when carrying out twice solid-liquid separation, and bits are a lot, and the inventive method requires that slag must be done, water
Divide 10% less than gross mass.
2, the inventive method adds laying particular stress on of Fructus Cannabis grouts mass fraction 1% in Fructus Cannabis grouts lysate
Sodium sulfite, can effectively prevent the oxidation of aldehydes matter in Fructus Cannabis grouts, it is ensured that last gained is low
The color of poly-Gly-His-Lys.
3, the inventive method adjusts pH value to 4.0, makes the albumen in Fructus Cannabis grouts be precipitated completely；Meanwhile,
Regulate with the HCl solution of 2.5mol/L, convenient control pH intensity of variation, the Urtica cannabina L. that the inventive method prepares
Core oligopeptide yield is high.
4, the inventive method Fructus Cannabis oligopeptide clear liquid uses membrance concentration, lyophilization, it is to avoid evaporation and concentration,
The too high generation causing Maillard reaction of spray drying temperature, improves the quality of Fructus Cannabis oligopeptide goods.
5, the inventive method utilizes Fructus Cannabis to extract remaining Fructus Cannabis grouts after Fructus Cannabis oil, by compound
The techniques such as protease hydrolyzed obtain Fructus Cannabis oligopeptide, improve the added value of fructus cannabis, improve simultaneously
The utilization rate of Fructus Cannabis.
The present invention uses through the Fructus Cannabis grouts that deoil of squeezing as raw material, in conjunction with isoelectric point, IP principle, precipitation
Albumen, then obtain Fructus Cannabis oligopeptide through compound protease enzymolysis and cryodesiccated method.Whole process profit
Use pure physical technology, substantially add without chemical composition, with low cost, environmentally safe.
Detailed description of the invention
Following instance is to further illustrate the present invention.
Embodiment 1
(1) put into after the Fructus Cannabis grouts 200kg (fat content is less than 1%) deoiled through squeezing being pulverized
5m3In extraction pot, described extraction pot is characterized by sandwich-type reacting by heating still, is used for dissolving Fructus Cannabis grouts powder.
In extraction pot, add 4 tons of pure water, adjust pH value to 9.0 by the NaOH solution of 2.5mol/L, add 2kg
Sodium metabisulfite, 70 DEG C of heated and stirred 30min, use automatic clinker-removing horizontal screw centrifuge 4000rpm to be centrifuged
20min, carries out solid-liquid separation, and clear liquid imports 5m3Settling tank, described settling tank is without reacting by heating still；With
The HCl solution of 2.5mol/L adjusts pH value to 4 protein precipitations, stirs, stands 2h, uses and automatically unload
Slag horizontal screw centrifuge carries out second time solid-liquid separation, abandons supernatant and stays slag charge, described slag charge to be Fructus Cannabis to divide
From albumen.
(2) after broken for separated protein powder, 5m is added3In extraction pot, by Fructus Cannabis separation albumen: pure water mass ratio
1:20 adds pure water, adjusts system temperature 48 DEG C, regulates pH to 8.5 by the NaOH solution of 2.5mol/L, presses
Adding compound protease according to the 3.0% of albumen gross mass, described compound protease quality proportioning is, pepsin:
Trypsin: flavor protease=4:3:3；Stirring enzymolysis 4h, heat up 80 DEG C of enzyme denaturing 20min, by tubular type from
Scheming is centrifuged to obtain clear liquid, and clear liquid imports 5m3Clear liquid storage tank.
(3) described clear liquid storage tank is connected with electrodialysis plant, and through electrodialysis, gained clear liquid is sloughed inorganic ions
After, import 5m3Refined storage tank.
(4) described refined storage tank is connected with membrance concentration equipment, uses saccharometer to record soluble solid content
Stop concentrating when accounting for quality of material 30%, import 2m3Concentrate storage tank.
(5) described concentration storage tank, is provided with discharging opening, and after discharging opening takes out, concentration feed liquid is carried out freezing
It is dried to obtain Fructus Cannabis oligomeric Gly-His-Lys 136.55kg.
The Fructus Cannabis oligomeric Gly-His-Lys color that the present embodiment prepares tends to white substantially, and non-metallic ion remains, and produces
Product yield 68%.Wherein kjeldahl determination fire detecting Fructus Cannabis oligomeric Gly-His-Lys total nitrogen content can reach 13.8%, trichloroacetic acid
Surveying oligopeptide content after precipitation high molecular weight protein and reach 78%, DNS method surveys contents of monosaccharides 4.8%, is high-quality
Fructus Cannabis oligopeptide.
Embodiment 2
(1) put into after the Fructus Cannabis grouts 200kg (fat content is less than 1%) deoiled through squeezing being pulverized
5m3In extraction pot, described extraction pot is characterized by sandwich-type reacting by heating still, is used for dissolving Fructus Cannabis grouts powder.
In extraction pot, add 4 tons of pure water, adjust pH value to 9.0 by 2.5mol/L NaOH solution, add 2kg inclined
Sodium bisulfite, 70 DEG C of heated and stirred 30min, use plate and frame type filter-press to carry out solid-liquid separation；Clear liquid imports
5m3Settling tank, described settling tank is without reacting by heating still, adjusts pH value to 4 precipitation eggs with 2.5mol/L solution
In vain, after stirring, stand 2h, use plate and frame type filter-press to carry out second time solid-liquid separation, abandon supernatant and stay
Slag charge, described slag charge is Fructus Cannabis separation albumen.
(2) after broken for separated protein powder, 5m is added3In extraction pot, by Fructus Cannabis separation albumen: pure water mass ratio
1:20 adds pure water, adjusts system temperature 48 DEG C, regulates PH to 8.5 by the NaOH solution of 2.5mol/L, presses
Adding compound protease according to the 3.0% of albumen gross mass, described compound protease quality proportioning is, pepsin:
Trypsin: flavor protease=4:3:3；Stirring enzymolysis 4h, heat up 80 DEG C of enzyme denaturing 20min, by tubular type from
Scheming is centrifuged to obtain clear liquid, and clear liquid imports 5m3Clear liquid storage tank.
(3) described clear liquid storage tank is connected with electrodialysis plant, and through electrodialysis, gained clear liquid is sloughed inorganic ions
After, import 5m3Refined storage tank.
(4) described refined storage tank is connected with membrance concentration equipment, uses saccharometer to record soluble solid content
Stop concentrating when accounting for quality of material 30%, import 2m3Concentrate storage tank.
(5) described concentration storage tank, is provided with discharging opening, and after discharging opening takes out, concentration feed liquid is carried out freezing
It is dried to obtain Fructus Cannabis oligomeric Gly-His-Lys 142.31kg.
The Fructus Cannabis oligomeric Gly-His-Lys color that the present embodiment prepares is faint yellow, and non-metallic ion remains, and product is received
Rate yield 71%.Wherein kjeldahl determination fire detecting Fructus Cannabis oligomeric Gly-His-Lys total nitrogen content can reach 12.4%, trichloroacetic acid
Surveying oligopeptide content after precipitation high molecular weight protein and reach 66%, DNS method surveys contents of monosaccharides 14.8%, is high-quality
Fructus Cannabis oligopeptide.
The above, the only present invention preferably detailed description of the invention, but protection scope of the present invention not office
Being limited to this, any those familiar with the art is in the technical scope of present disclosure, according to this
The technical scheme of invention and inventive concept thereof in addition equivalent or change, all should contain the protection in the present invention
Within the scope of.
Claims (4)
1. the method extracting Fructus Cannabis oligopeptide from Fructus Cannabis grouts, it is characterised in that concrete steps
For:
S1, the collection fat content Fructus Cannabis grouts less than 1%, by Fructus Cannabis grouts: the mass ratio of water=1:20
In described Fructus Cannabis grouts, add water, adjust pH to 9.0 by the NaOH solution of 2.5mol/L, add described
The sodium metabisulfite of Fructus Cannabis grouts quality 1% amount, carries out solid-liquid for the first time after 70 DEG C of heated and stirred 30min
Separate, collect liquid phase；In described liquid phase, add the HCl solution of 2.5mol/L, regulate pH to 3.8～4.3
After, stand 2h and carry out second time solid-liquid separation, collect precipitation, be Fructus Cannabis separation albumen；
S2, add water in the Fructus Cannabis separation albumen that step S1 prepares, described Fructus Cannabis separation albumen and water
Mass ratio is 1:15～25；It is heated to 40～55 DEG C, regulates pH to 7.0～9.0 by the NaOH solution of 2.5mol/L,
Add 1.0～5.0% compound protease of amount of described Fructus Cannabis separation albumen quality；After stirring enzymolysis 3～6h,
It is warming up to 80～95 DEG C of enzyme denaturing 10～25min, centrifugal collection clear liquid, it is Fructus Cannabis oligopeptide solution；
The quality proportioning of described compound protease is: pepsin: trypsin: flavor protease
=3～5:2～4:2～4；
S3, the Fructus Cannabis oligopeptide solution that step S2 prepares is carried out electrodialysis, is sloughed inorganic ions after, warp
Membrance concentration equipment concentrates, and obtains Fructus Cannabis oligopeptide concentrated solution；In described Fructus Cannabis oligopeptide concentrated solution solvable
Property solid content is 20～40wt%；By described Fructus Cannabis oligopeptide concentrated solution lyophilization, obtain Fructus Cannabis
Oligopeptide.
The method extracting Fructus Cannabis oligopeptide the most according to claim 1 from Fructus Cannabis grouts, its feature
Being, solid-liquid separation described in step S1 uses automatic discharge horizontal screw centrifuge 4000rpm to be centrifuged.
The method extracting Fructus Cannabis oligopeptide the most according to claim 1 from Fructus Cannabis grouts, its feature
Being, described in step S3, electrodialytic operating condition is: voltage 75V, enters Fructus Cannabis oligopeptide solution flow rate
1.55 ton hour, concentrated stream speed 1.60 ton hour, cooling of electrode water flow velocity 1 ton hour.
The method extracting Fructus Cannabis oligopeptide the most according to claim 1 from Fructus Cannabis grouts, its feature
Being, membrance concentration equipment described in step S3 uses hollow fiber nanofiltration membrane, and operation pressure is 1.5～1.6Mpa.
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Cited By (11)
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| CN107699600A (en) * | 2017-11-01 | 2018-02-16 | 云南谷益美农业开发有限公司 | The method that fructus cannabis oligopeptide is extracted from fructus cannabis grouts |
| US10493377B1 (en) | 2019-02-06 | 2019-12-03 | Heinkel Filtering Systems, Inc. | Biomass extraction and centrifugation systems and methods |
| CN110777183A (en) * | 2019-12-04 | 2020-02-11 | 山西原生肽科技有限公司 | Fructus cannabis oligopeptide powder and preparation method and application thereof |
| CN111345356A (en) * | 2020-04-27 | 2020-06-30 | 上海来伊份股份有限公司 | Preparation method of high-fibrin meat embryo |
| US10751640B1 (en) | 2019-10-30 | 2020-08-25 | Heinkel Filtering Systems, Inc. | Cannabidiol isolate production systems and methods |
| US10765966B2 (en) | 2019-02-06 | 2020-09-08 | Heinkel Filtering Systems. Inc. | Biomass extraction and centrifugation systems and methods |
| US10858303B1 (en) | 2019-10-30 | 2020-12-08 | Heinkel Filtering Systems, Inc. | Cannabidiol isolate production systems and methods |
| CN112410393A (en) * | 2020-11-30 | 2021-02-26 | 中国农业科学院麻类研究所 | Cannabis bioactive peptide and preparation method and application thereof |
| CN112402415A (en) * | 2020-11-30 | 2021-02-26 | 湖北瑞邦生物科技有限公司 | Application of fructus cannabis oligopeptide |
| CN114381486A (en) * | 2022-01-20 | 2022-04-22 | 厦门元之道生物科技有限公司 | Fructus cannabis functional polypeptide and preparation method thereof |
| CN115251403A (en) * | 2022-07-15 | 2022-11-01 | 中国农业科学院麻类研究所 | Muscle-increasing nutritional composition containing fructus cannabis peptide and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107699600A (en) * | 2017-11-01 | 2018-02-16 | 云南谷益美农业开发有限公司 | The method that fructus cannabis oligopeptide is extracted from fructus cannabis grouts |
| US10493377B1 (en) | 2019-02-06 | 2019-12-03 | Heinkel Filtering Systems, Inc. | Biomass extraction and centrifugation systems and methods |
| US10765966B2 (en) | 2019-02-06 | 2020-09-08 | Heinkel Filtering Systems. Inc. | Biomass extraction and centrifugation systems and methods |
| US10858303B1 (en) | 2019-10-30 | 2020-12-08 | Heinkel Filtering Systems, Inc. | Cannabidiol isolate production systems and methods |
| US10751640B1 (en) | 2019-10-30 | 2020-08-25 | Heinkel Filtering Systems, Inc. | Cannabidiol isolate production systems and methods |
| CN110777183A (en) * | 2019-12-04 | 2020-02-11 | 山西原生肽科技有限公司 | Fructus cannabis oligopeptide powder and preparation method and application thereof |
| CN111345356A (en) * | 2020-04-27 | 2020-06-30 | 上海来伊份股份有限公司 | Preparation method of high-fibrin meat embryo |
| CN111345356B (en) * | 2020-04-27 | 2023-04-18 | 上海来伊份股份有限公司 | Preparation method of high-fibrin meat embryo |
| CN112410393A (en) * | 2020-11-30 | 2021-02-26 | 中国农业科学院麻类研究所 | Cannabis bioactive peptide and preparation method and application thereof |
| CN112402415A (en) * | 2020-11-30 | 2021-02-26 | 湖北瑞邦生物科技有限公司 | Application of fructus cannabis oligopeptide |
| CN112402415B (en) * | 2020-11-30 | 2022-05-06 | 湖北瑞邦生物科技有限公司 | Application of fructus cannabis oligopeptide |
| CN114381486A (en) * | 2022-01-20 | 2022-04-22 | 厦门元之道生物科技有限公司 | Fructus cannabis functional polypeptide and preparation method thereof |
| CN114381486B (en) * | 2022-01-20 | 2023-09-15 | 厦门元之道生物科技有限公司 | Fructus cannabis functional polypeptide and preparation method thereof |
| CN115251403A (en) * | 2022-07-15 | 2022-11-01 | 中国农业科学院麻类研究所 | Muscle-increasing nutritional composition containing fructus cannabis peptide and preparation method thereof |
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Effective date of registration: 20191106 Address after: 034000 Shanxi province Xinzhou Xinfu District road construction Yi Ju B10 District Court Building 2 unit 1 West households Applicant after: Shanxi Han Ma biological science and Technology Co., Ltd. Address before: 12, 116000 floor, No. 541, Huangpu Road, Ganjingzi District, Liaoning, Dalian, China (1207-16) Applicant before: Dalian Deep Blue Peptide Technology Research AND Development Co., Ltd. |
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Application publication date: 20161012 |