CN109536556A - A kind of preparation method of homogeneity green alga active peptide - Google Patents
A kind of preparation method of homogeneity green alga active peptide Download PDFInfo
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- CN109536556A CN109536556A CN201811629177.2A CN201811629177A CN109536556A CN 109536556 A CN109536556 A CN 109536556A CN 201811629177 A CN201811629177 A CN 201811629177A CN 109536556 A CN109536556 A CN 109536556A
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- green alga
- active peptide
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- 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
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/10—Process efficiency
Abstract
The invention discloses a kind of preparation methods of homogeneity green alga active peptide, belong to natural active matter extractive technique field.The present invention prepares green alga active peptide the following steps are included: (1) use green alga enteromorpha powder for raw material, controls key process parameters such as temperature, solid-to-liquid ratio, temperature, pH, passes through counterflow extraction technique extraction green alga albumen；(2) said extracted liquid is carried out digesting to obtain enzymolysis liquid using compound protease；(3) by enzymolysis liquid obtained above using ultrafiltration membrane purifying concentration, high temperature enzyme deactivation obtains algal active peptide liquid；(4) green alga active peptide powder is obtained by spray drying.A kind of preparation method of green alga active peptide of the invention, extracted by establishing multi-stage countercurrent-digest barrier separation integral process, recovery rate is high, can continuous production, bioactive peptide molecule amount it is uniform controllable, residue, filtrate after extracting albumen can be used as raw material for producing alga fertilizer.
Description
Technical field
The invention belongs to natural active matter extractive technique fields, and in particular to a kind of preparation of homogeneity green alga active peptide
Method.
Background technique
Multistage countercurrent solid-liquid extractive technique is a kind of solid-phase material and solvent direction of relative movement is opposite, continuous and quantitative is added
Solid-phase material and solvent and the continuous separation technology for exporting residue and extracting solution.In the extraction process of effective component, solid
It is in opposite direction flowing with liquid, the solvent containing effective component is contacted before outflow system with fresh raw material, and is passed through multiple
The raw material of extraction is contacted before outflow system with fresh solvent, so that it is complete to extract effective component.With traditional extraction side
For method compared to having the significant advantage that (1) effective component leaching rate is fast, product yield is high；(2) can continuous work, be suitable for big
Large-scale production；(3) solvent usage is saved, Steep cencentration is high, while reducing post-treatment operations；It (4) can effectively save energy
Source, yield are big；(5) Extracting temperature needed for is low, and the extraction for the material for being both suitble to thermal stability good is also applied for heat-sensitive substance
It extracts.To sum up, it is feasible for multi-stage countercurrent extraction process being prepared active peptide.
Since two thousand seven, Huang Bohai Offshore in China's breaks out Enteromorpha green tide year after year, to sea transport, sea-farming and tourism
Sightseeing causes to seriously affect.Green alga Enteromorpha be rich in multiple nutritional components, as polysaccharide, albumen, unsaturated fatty acid, vitamin and
Minerals etc..Marine algae extract is relatively confined to algal polysaccharides, seaweed glue class product, economic benefit and resource utilization at present
It is low.The research history of algal active peptide is shorter, and most of research should concentrate on Soyprotein peptide, fish protein peptide etc., is mainly used for
It reduces cholesterol, blood pressure lowering and improves the immunity of the human body.Algal active peptide because of its special bioactivity, increasingly by
The concern of scientist, China are less for algal active peptide Research foundation, and industrialization product is less.Patent 201610488495.6
A kind of method that oligopeptide production marine alga oligopeptide powder is extracted from seaweed is disclosed, specific steps include cleaning seaweed, shining
It is dry, it crushes, obtains seawood meal；Add water, defibrination；Seaweed slurries are heated, pH to 3.3~4.0 is adjusted, pectase, stirring is added
Enzymatic hydrolysis；NaOH is added, adjusts pH to 7.0~9.0, compound protease, stirring enzymatic hydrolysis is added；Heat up enzyme deactivation, is separated by solid-liquid separation, and collects
Liquid phase；Clear liquid is handled using membrane filtering method, collects permeate；It will transmit through liquid and carry out electrodialysis, absorbent charcoal fine purification, de- charcoal, steam
Hair is concentrated, is dry that marine alga oligopeptide powder, the molecular weight of gained small peptide are less than 2000Da mostly.The comprehensive analysis prior art,
In order to improve the recovery rate of seaweed polypeptide, Sargassum protein is mostly degraded to small peptide using enzymatic hydrolysis or complex enzyme hydrolysis technique, it is mostly low
In 2000Da, and the standby shortage accuracy control technology of small peptide beam system preferable for antioxidant activity, the homogeneity of product is poor,
Therefore, the key technology technology of high anti-oxidation active alga small peptide (such as molecular weight 3000-4000Da) is established, green alga small peptide is improved
Homogeneity become the industry field urgent need to resolve the technical issues of.
Summary of the invention
In view of the deficiencies of the prior art, it is an object of that present invention to provide a kind of preparation method of homogeneity green alga active peptide,
Extract-digest barrier separation integral process by the multi-stage countercurrent of foundation and produce green alga active peptide, recovery rate is high, can continuous production,
Bioactive peptide molecule amount is uniform controllable, and oxidation resistance is strong；Residue, filtrate after extracting albumen can be used as raw material for producing seaweed
Fertilizer.
To achieve the above object, the present invention uses following technological means:
A kind of preparation method of homogeneity green alga active peptide of the present invention, specifically includes the following steps:
(1) using chlorella powder as raw material, using counterflow extraction technique, solid-to-liquid ratio 1:5-1:10, temperature 40-60 are maintained
DEG C, pH9-11 extracts green alga albumen；
(2) said extracted liquid is used into protease hydrolyzed 3-6h, obtains green alga protein enzymatic hydrolyzate；
(3) ultrafiltration membrane for being 3KDa and 4KDa by molecular weight by above-mentioned enzymolysis liquid obtains concentrate and high temperature enzyme deactivation；
(4) enzymolysis liquid is spray-dried, it is green to get arriving to divide Han Liang≤10% by the Shui of active peptide after control spray drying
Algae active peptide powder, wherein the inlet air temperature being spray-dried is 180-200 DEG C, flow velocity 2-5ml/min.
Extraction solvent is sodium hydroxide weak caustic solution in above-mentioned steps (1), and adds 1% cellulase, 0.5% pectin
Enzyme.
Preferably, said extracted green alga albumen uses five stage countercurrent extractive technique of pot group type, maintains solid-to-liquid ratio 1:8, temperature
55 DEG C, pH10.
Protease described in above-mentioned steps (2) is compound protease, and additive amount is the 3% of green alga mass ratio；Compound egg
The group of white enzyme becomes papain and subtilopeptidase A, mass ratio 2:1.
Preferably, above-mentioned steps (2) digest 4h using compound protease, and molecular weight tends towards stability, using Ultra filtration membrane
Afterwards, polypeptide stable homogeneous can be improved.
Preferably, green alga protein enzymatic hydrolyzate is heated to 90 DEG C of heat preservation 15min enzyme deactivation sterilizings in the step (3).
The present patent application protects the green alga active peptide being prepared according to above-mentioned preparation method.Small peptide content be 90% with
On, molecular weight 3-4KDa.
Compared with prior art, the present invention it is advantageous that:
1) barrier separation integral process is extracted-digests using multi-stage countercurrent, product yield height is up to 90% or more, with polysaccharide point
From rate up to 95% or more, can continuous work, molecular weight is uniform controllable, and uniformity is high；
2) unlike the prior art, extraction process uses pot group type counterflow extraction technique, adds in Extraction solvent
Enter a certain amount of cellulase and pectase, effective component leaching rate is fast, and product yield is high, can save solvent usage, can connect
Continuous work, is suitble to large-scale production；
3) it is prepared using papain and subtilopeptidase A combined-enzyme method, reaction condition is mild, and enzymolysis process is easy
In control, residue, filtrate after extracting albumen can be used as raw material and be used to produce alga fertilizer, environmentally friendly；
4) present invention extracts-digests barrier separation integral process technology production green alga active peptide by establishing multi-stage countercurrent, gram
It has taken in the prior art for the technology drawback for improving Chlorella polypeptide recovery rate and polypeptide of thoroughly degrading, the application had both realized green
The high extraction of algae polypeptide, and obtained the Chlorella polypeptide that molecular weight homogeneity is strong, functional activity is good.
Specific embodiment
The technical scheme of the present invention will be explained in further detail With reference to embodiment.
The preparation of 1 Enteromorpha active peptide of embodiment
A kind of preparation method of Enteromorpha active peptide, specifically includes the following steps:
(1) it uses enteromorpha powder for raw material, using 5 stage countercurrent extractive technique of pot group type, maintains solid-to-liquid ratio 1:8,55 DEG C of temperature,
PH10 extracts green alga albumen；Extraction solvent is sodium hydroxide weak caustic solution, and adds 1% cellulase, 0.5% pectase；
(2) above-mentioned protein liquid is used into papain and subtilopeptidase A complex enzyme hydrolysis 4h, obtains enteromorpha protein enzymatic hydrolysis liquid；On
3% that complex enzyme additive amount is green alga mass ratio is stated, wherein papain and subtilopeptidase A, mass ratio 2:1；
(3) ultrafiltration membrane for being 3KDa and 4KDa by molecular weight by said extracted liquid, obtains concentrate, is heated to 90 DEG C of heat preservations
15min enzyme deactivation；
(4) enzymolysis liquid is spray-dried, the Shui of active peptide divides Han Liang≤10% to get waterside is arrived after control spray drying
Tongue fur active peptide powder, wherein the inlet air temperature being spray-dried is 200 DEG C, flow velocity 3ml/min；
Through detecting, in Enteromorpha active peptide obtained by the present embodiment, recovery rate 92.4%, enzymatic hydrolyzation 97.2% is small
Peptide content is up to 95% or more, molecular weight 3.25KDa.
The preparation method of 2 sea lettuce active peptide of embodiment
A kind of preparation method of sea lettuce active peptide, specifically includes the following steps:
(1) it uses ulva lactuca powder for raw material, using 5 stage countercurrent extractive technique of pot group type, maintains solid-to-liquid ratio 1:8,55 DEG C of temperature,
PH10 extracts green alga albumen；Extraction solvent is sodium hydroxide weak caustic solution, and adds 1% cellulase, 0.5% pectase；
(2) above-mentioned protein liquid is used into papain and subtilopeptidase A complex enzyme hydrolysis 4h, obtains sea lettuce albumen
Enzymolysis liquid；Above-mentioned complex enzyme additive amount is the 3% of green alga mass ratio, wherein papain and subtilopeptidase A, quality
Than for 2:1；
(3) ultrafiltration membrane for being 3KDa and 4KDa by molecular weight by above-mentioned enzymolysis liquid, obtains concentrate, is heated to 90 DEG C of heat preservations
15min enzyme deactivation；
(4) enzymolysis liquid is spray-dried, the Shui of active peptide divides Han Liang≤10% to get stone is arrived after control spray drying
Water shield active peptide powder, wherein the inlet air temperature being spray-dried is 200 DEG C, flow velocity 3ml/min；
Through detecting, in sea lettuce active peptide obtained by the present embodiment, recovery rate 93.7%, enzymatic hydrolyzation 97.8% is small
Peptide content is up to 95% or more, molecular weight 3.41KDa.
The test of 3 active peptide oxidation resistance of embodiment
Examination is compared using green alga active peptide preparation-obtained in embodiment 1,2 as antioxidant and ascorbic acid
It tests, measures it and remove DPPH, ABTS, hydroxyl radicals, test result is shown in Table 1:
The evaluation of 1 green alga active peptide antioxidant activity of table
As seen from the above table, embodiment 3 is used for using embodiment 1,2 preparation-obtained green alga active peptides as antioxidant
Experiment, shows that green alga active peptide of the present invention can effectively remove DPPH, ABTS, hydroxy radical, has apparent antioxidant activity.
Influence of 4 Different Preparation of embodiment to green alga activity peptides extraction rate
Carry out influence of the Different Preparation to green alga activity peptides extraction rate and antioxidant activity.Wherein control group without
Compound protein enzymatic hydrolysis directly carries out film filtering, spray drying, remaining technique is the same as embodiment 1；Comparative examples 1 use Papain
The single enzymatic hydrolysis of enzyme, remaining process conditions is the same as embodiment 1；Comparative examples 2 use the single enzymatic hydrolysis of subtilopeptidase A, remaining work
Skill condition is the same as embodiment 1；Experimental result is as shown in table 2:
Influence of 2 Different Preparation of table to green alga activity peptides extraction rate
| Group | Embodiment 1 | Control group | Comparative examples 1 | Comparative examples 2 |
| Recovery rate (%) | 92.4 | 28.3 | 68.3 | 42.7 |
The complex enzyme hydrolysis that the application uses as shown in Table 2 has been obviously improved the recovery rate of Chlorella polypeptide, illustrates Papain
Enzyme and subtilopeptidase A have significant synergistic function.In addition, test result shows during the invention
The recovery rate for matching comparison active peptide and antioxidant activity of papain and subtilopeptidase A all have significant impact
(p<0.01)。
The above examples are only used to illustrate the technical scheme of the present invention, rather than is limited；Although referring to aforementioned reality
It applies example to be described in detail to by invention, but for those of ordinary skill in the art, it still can be to aforementioned reality
Technical solution documented by example is applied to modify or equivalent replacement of some of the technical features；And to these modifications
Or replacement, the spirit and scope for claimed technical solution of the invention that it does not separate the essence of the corresponding technical solution.
Claims (7)
1. a kind of preparation method of homogeneity green alga active peptide, it is characterised in that specifically includes the following steps:
(1) using chlorella powder as raw material, using counterflow extraction technique, solid-to-liquid ratio 1:5-1:10 is maintained, 40-60 DEG C of temperature,
PH9-11 extracts green alga albumen；
(2) said extracted liquid is used into protease hydrolyzed 3-6h, obtains green alga protein enzymatic hydrolyzate；
(3) ultrafiltration membrane for being 3KDa and 4KDa by molecular weight by above-mentioned enzymolysis liquid obtains concentrate and high temperature enzyme deactivation；
(4) enzymolysis liquid is spray-dried, the Shui of active peptide divides Han Liang≤10% to get living to green alga after control spray drying
Property peptide powder, wherein the inlet air temperature being spray-dried be 180-200 DEG C, flow velocity 2-5ml/min.
2. a kind of preparation method of homogeneity green alga active peptide according to claim 1, it is characterised in that the step (1)
The solvent that middle multi-stage countercurrent extracts is sodium hydroxide weak caustic solution, and adds 1% cellulase, 0.5% pectase.
3. a kind of preparation method of homogeneity green alga active peptide according to claim 1, it is characterised in that the multistage is inverse
Stream extract green alga albumen use five stage countercurrent extractive technique of pot group type, maintain solid-to-liquid ratio 1:8,55 DEG C of temperature, pH10.
4. a kind of preparation method of homogeneity green alga active peptide according to claim 1, it is characterised in that the step (2)
Described in protease be compound protease, additive amount be green alga mass ratio 3%；The group of compound protease becomes Papain
Enzyme and subtilopeptidase A, mass ratio 2:1.
5. a kind of preparation method of homogeneity green alga active peptide according to claim 1, it is characterised in that the step (2)
4h is digested using compound protease.
6. a kind of preparation method of homogeneity green alga active peptide according to claim 1, it is characterised in that the step (3)
It is middle that green alga protein enzymatic hydrolyzate is heated to 90 DEG C of heat preservation 15min enzyme deactivation sterilizings.
7. the green alga active peptide being prepared by preparation method described in claim 1-6, small peptide content is 90% or more, molecular weight
3-4KDa。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110106218A (en) * | 2019-04-24 | 2019-08-09 | 北海生巴达生物科技有限公司 | A kind of algae small-molecular peptides extracting method |
| CN114395607A (en) * | 2022-03-28 | 2022-04-26 | 卡力(烟台)农业发展有限公司 | Seaweed oligopeptide powder and preparation method thereof |
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2018
- 2018-12-28 CN CN201811629177.2A patent/CN109536556A/en not_active Withdrawn
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110106218A (en) * | 2019-04-24 | 2019-08-09 | 北海生巴达生物科技有限公司 | A kind of algae small-molecular peptides extracting method |
| CN114395607A (en) * | 2022-03-28 | 2022-04-26 | 卡力(烟台)农业发展有限公司 | Seaweed oligopeptide powder and preparation method thereof |
| CN114395607B (en) * | 2022-03-28 | 2022-07-12 | 卡力(烟台)农业发展有限公司 | Seaweed oligopeptide powder and preparation method thereof |
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Application publication date: 20190329 |