CN104774827A - Method for preparing alginate lyase from abalone internal organs - Google Patents
Method for preparing alginate lyase from abalone internal organs Download PDFInfo
- Publication number
- CN104774827A CN104774827A CN201510124968.XA CN201510124968A CN104774827A CN 104774827 A CN104774827 A CN 104774827A CN 201510124968 A CN201510124968 A CN 201510124968A CN 104774827 A CN104774827 A CN 104774827A
- Authority
- CN
- China
- Prior art keywords
- algin catenase
- preparation
- algin
- add
- obtains
- 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
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y402/00—Carbon-oxygen lyases (4.2)
- C12Y402/02—Carbon-oxygen lyases (4.2) acting on polysaccharides (4.2.2)
- C12Y402/02003—Poly(beta-D-mannuronate) lyase (4.2.2.3)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y402/00—Carbon-oxygen lyases (4.2)
- C12Y402/02—Carbon-oxygen lyases (4.2) acting on polysaccharides (4.2.2)
- C12Y402/02011—Poly(alpha-L-guluronate) lyase (4.2.2.11), i.e. alginase II
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention provides a method for preparing alginate lyase from abalone internal organs. The method comprises the following steps: performing crude enzyme extraction; processing the extract by active carbon, filtering to remove the active carbon to obtain a filtrate; subjecting the filtrate to anion exchange chromatography; carrying out a membrane treatment; and finally drying the condensate through spraying or freezing to obtain an alginate lyase product with a high purity and high activity. The active carbon is used to remove the color and odor, the ion exchange chromatography is used to remove the impurity proteins; and technologies such as membrane condensation, spray drying, freezing drying, and the like are adopted so as to prepare high-purity and high-activity alginate lyase from abalone internal organs. The provided method can be amplified and applied to the industrial production, the technology is simple, and the separation effect is good. At the same time, the production cost is saved, the technology can be amplified, and the application prospect is good.
Description
Technical field
The present invention relates to marine prods manufacture field, more specifically relate to a kind of preparation method of algin catenase.
Background technology
Algin catenase is that a class can hydrolysis of brown algae glue, forms the enzyme of algin oligosaccharide.Algin oligosaccharide, owing to having special chemical property and biological activity, becomes a kind of potential functional product in recent years and constantly receives publicity.And the focus that enzyme-algin catenase that algin changes algin oligosaccharide into also becomes research at present can be realized.The linear polysaccharide that algin is made up of α-L-guluronic acid and beta-D-mannuronic acid.Poly guluronic acid (PG), polymannuronic acid (PM) and irregular hybrid fragments PMG can be divided into by polymerized form.PG lyase, PM lyase and PMG lyase is divided into according to the fracture mode difference of algin catenase in algin.Algin catenase be mainly present in thalassiomycetes and with marine algae be food Organisms in.Theoretical and to be applied in research both at home and abroad very few for it.Study hotspot main is at present the functionally active determining algin catenase enzymolysis product algin oligosaccharide.Its major function activity mainly contains: the division promoting vegetable cell, and then the growth promoting plant; Promote human endothelial cells and Keratinocytic growth, there is water retention characteristic, develop as cosmetics; Immunity moderation function, strengthening immunity, the effects such as anti-bacteria and anti-virus.
According to China Fisheries yearbook statistic data, within 2012, China's abalone output is 9.1 ten thousand tons, comparatively within 2011, increases by 18.11%, and presents the trend increased year by year.Abalone take seaweeds as food, containing abundant polysaccharide hydrolysis enzyme in its digestive tube, as: cellulase, algin catenase, agarase etc.In the abalone course of processing, a large amount of processing fent-internal organ of generation, often by reject or be prepared into the low value products such as fish meal, not only bring serious environmental pollution, and cause the waste of resource.Therefore, with abalone processing byproduct internal organ for raw material efficiently prepares algin catenase, effectively can improve the added value of abalone processing, reduce the environmental pollution caused because of internal organ are directly discarded.
The separation and purification about algin catenase of bibliographical information so far mainly adopts the method that ammonium sulfate precipitation and multistep column chromatography combine, separation and purification process is loaded down with trivial details, complicated operation, length consuming time, yield are low, cost is high, are difficult to realize suitability for industrialized production.
Summary of the invention
The object of the present invention is to provide that a kind of technique is simple, good separating effect, the preparation method of the abalone internal organ algin catenase of suitability for industrialized production can be realized.
For achieving the above object, the invention provides a kind of with abalone internal organ for the method for algin catenase prepared by raw material, it is characterized in that comprising the steps:
1) thick enzyme extraction: shredded by abalone internal organ, add Tris-HCl damping fluid, tissue mashing, centrifugal, supernatant liquor is crude enzyme liquid;
2) activated carbon treatment: described crude enzyme liquid is crossed and filtered gac after activated carbon treatment, and gained is filtrate;
3) anion exchange chromatography: by gained filtrate through DEAE-Sepharose, DEAE-Sephacel or Q-Sepharose anionite-exchange resin, the resin after absorption, through stepwise elution desorb, obtains the algin catenase of higher degree;
4) film process 1: by the ultra-filtration membrane ultrafiltration removal of impurities having algin catenase active part leacheate 100-50 kDa molecular weight to retain, obtain highly purified algin catenase;
5) by ultra-filtration membrane ultrafiltration and concentration that outer for above-mentioned ultrafiltration liquid part 10-3 kDa molecular weight retains;
6) enzyme preparation: by spray-dried for described concentrated solution or lyophilize, obtains the algin catenase goods of high purity, high vigor;
Optional, in described concentrated solution, add trehalose or beta-cyclodextrin before described spraying dry.
In described step 1), add the Tris-HCl damping fluid of 4 times of volume 20 mmol/L pH 8.5; Described centrifugal be centrifugal 20 min of 12000 × g.
Described step 2) in, described gac add-on is that (W/V) 0.5-3 %(refers to gac (w) and accounts for crude enzyme liquid ratio (v)), processing mode is 4 DEG C of stir process 30-60 min.
In described step 3), described stepwise elution desorb be absorption after resin through 0.05,0.1,0.2,0.3 mol/L NaCl salt concn stepwise elution.
In described step 5), the add-on of described trehalose or beta-cyclodextrin is 10-30 bulking value %.
The present invention removes a large amount of pigments by charcoal absorption, reaches the effect of de-raw meat decolouring simultaneously; Utilize anion exchange chromatography can by algin catenase active adsorption, by changing salt concn stepwise elution mode, algin catenase is well separated with foreign protein (as: cytoskeletal protein, cellulase, the albumen that kethepsin equal size is higher).The present invention is after ion-exchange chromatography, adopt ultra-filtration membrane that algin catenase is carried out to ultrafiltration removal of impurities, concentrates, both achieve the purifying of algin catenase, again can by sample concentration to smaller size smaller, facilitate subsequent spray drying or freezing dry process, save the energy.In spray-drying process, in enzyme liquid, add the protective material such as trehalose or beta-cyclodextrin, protect the activity of enzyme.
In sum, the present invention adopts activated carbon decolorizing to take off raw meat, ion-exchange chromatography is except foreign protein, and the technology such as membrane filtration removal of impurities, spraying dry obtain the algin catenase that purity is high, vigor is good from abalone internal organ, and this technique can be applicable to suitability for industrialized production after amplifying.
The present invention, by charcoal absorption, eliminates chromoprotein, the polysaccharide of part; And then employing ion-exchange chromatography, operational phase type of elution removes a large amount of foreign proteins, simplifies purifying process, obtains highly purified algin catenase.Present invention, avoiding adopt in prior art ammonium sulfate precipitation, multistep column chromatography combine method, simplify technical process, saved production cost, can technology scale amplification be carried out, have a good application prospect.
Accompanying drawing explanation
Fig. 1 is the SDS-PAGE electrophorogram of purifying algin catenase of the present invention.
Fig. 2 is sequence and other species algin catenase sequence alignments figure of the acquisition of preparation-obtained peptide mass fingerprinting spectrum.
Fig. 3 is to the discomposing effect of algin and viscosity profile by the algin catenase of preparation.
Fig. 4 is to the decomposition viscosity profile of algin by the algin catenase of preparation.
Embodiment
Be described below in detail embodiments of the invention, the example of described embodiment is shown in the drawings, and wherein same or similar label represents same or similar element or has element that is identical or similar functions from start to finish.Be exemplary below by the embodiment be described with reference to the drawings, be intended to for explaining the present invention, and can not limitation of the present invention be interpreted as.Unreceipted concrete technology or condition person in embodiment, according to the technology described by the document in this area or condition or carry out according to product description.Agents useful for same or the unreceipted production firm person of instrument, being can by the conventional products of commercial acquisition.
embodiment 1: the preparation of algin catenase
1. thick enzyme extraction: get 150 g abalone internal organ, chopping, add 600 mL volume 20 mmol/L Tris-HCl (pH 8.5), tissue mashing, centrifugal 20 min of 12000 × g, get supernatant, obtain 650 mL crude enzyme liquids;
2. activated carbon treatment: add 3.25 g gacs in 650 mL crude enzyme liquids, whip attachment 30 min at 4 DEG C.Carry out suction filtration with filter paper after absorption, obtain 630 mL filtrates;
3. anion exchange chromatography: above-mentioned filtrate after charcoal absorption is splined on the DEAE-Sepharose anion-exchange column (2.5 × 10 cm) balanced with 20 mmol/L Tris-HCl (pH 8.5), flow velocity 1.0 mL/min.The absorbance of effluent liquid at 280 nm places is washed till to baseline with the abundant stream of 20 mmol/L Tris-HCl (pH 8.5) after completion of the sample.Then stepwise elution is carried out respectively with 20 mmol/L Tris-HCl (pH 8.5) damping fluid containing 0.05 mol/L, 0.1 mol/L, 0.2 mol/L, 0.3 mol/L NaCl, collect the algin catenase that 0.1 mol/L NaCl leacheate (5 mL/ pipe) obtains higher degree, totally 400 mL;
4. film process 1: contain the 50 kDa ultra-filtration membrane ultrafiltration of highly purified algin catenase elutriant by what obtain, obtains the outer liquid of 360 mL ultrafiltration;
5. film process 2: outer for above-mentioned ultrafiltration cup liquid 3 kDa ultra-filtration membranes are concentrated, obtains concentrated solution 50 mL;
6. enzyme preparation: 50 mL concentrated solutions are carried out lyophilize, obtains zymin.
embodiment 2: the preparation of algin catenase
1. thick enzyme extraction: get 600 g abalone internal organ, chopping, add 2.4 L 20 mmol/L Tris-HCl (pH 8.5), tissue mashing, centrifugal 20 min of 12000 × g, get supernatant, obtain 2.5 L crude enzyme liquids;
2. activated carbon treatment: add 75 g gacs in 2.5 L crude enzyme liquids, whip attachment 45 min at 4 DEG C.Carry out suction filtration with filter paper after absorption, obtain 2.4 L filtrates;
3. anion exchange chromatography: be splined on through above-mentioned filtrate after charcoal absorption the DEAE-Sephacel anion-exchange column (5 × 15 cm) balanced with 20 mmol/L Tris-HCl (pH 8.5), flow velocity 4.0 mL/min.The absorbance of effluent liquid at 280 nm places is washed till to baseline with the abundant stream of 20 mmol/L Tris-HCl (pH 8.5) after completion of the sample.Then stepwise elution is carried out respectively with 20 mmol/L Tris-HCl (pH 8.5) damping fluid containing 0.05 mol/L, 0.1 mol/L, 0.2 mol/L, 0.3 mol/L NaCl, collect the algin catenase that 0.1 mol/L NaCl leacheate (30 mL/ pipe) obtains higher degree, totally 2.2 L;
4. film process 1: contain the 50 kDa ultra-filtration membrane ultrafiltration of highly purified cellulase elutriant by what obtain, obtains the outer liquid of 2.0 L ultrafiltration;
5. film process 2: outer for above-mentioned ultrafiltration cup liquid 10 kDa ultra-filtration membranes are concentrated, obtains concentrated solution 300 mL;
6. enzyme preparation: add 30g beta-cyclodextrin and carry out spraying dry in concentrated solution, obtain zymin.
embodiment 3: the preparation of algin catenase
1. enzyme liquid extracts: get 3 kg abalone internal organ, chopping, and add 12 L 20 mmol/L Tris-HCl (pH 8.5), tissue mashing, centrifugal 20 min of 12000 × g, get supernatant, obtain 12.6 L crude enzyme liquids;
2. charcoal absorption: add 300 g gacs in 12.6 L crude enzyme liquids, whip attachment 60 min at 4 DEG C.Adsorb rear filter-cloth filtering, obtain 12.4 L filtrates;
3. Anionic column chromatography: be splined on through above-mentioned filtrate after charcoal absorption the Q-Sepharose anion-exchange column (5 × 30 cm) balanced with 20 mmol/L Tris-HCl (pH 8.5), flow velocity 20.0 mL/min.The absorbance of effluent liquid at 280 nm places is washed till to baseline with the abundant stream of 20 mmol/L Tris-HCl (pH 8.5) after completion of the sample.Then stepwise elution is carried out with 20 mmol/L Tris-HCl (pH 8.5) damping fluid containing 0.05 mol/L, 0.1 mol/L, 0.2 mol/L, 0.3 mol/L NaCl, collect 0.2 mol/L NaCl leacheate and obtain highly purified algin catenase, totally 8.4 L;
4. film process 1: by using containing highly purified algin catenase elutriant of obtaining: 100 kDa ultra-filtration membrane ultrafiltration, obtains the outer liquid of 8.2 L ultrafiltration;
5. film process 2: outer for above-mentioned ultrafiltration cup liquid 10 kDa ultra-filtration membranes are concentrated, obtains concentrated solution 1.6 L;
6. enzyme preparation: add 480 g trehaloses and carry out spraying dry in concentrated solution, obtain 465 g zymins.
embodiment 4: the proof test of algin catenase
In order to verify the molecular weight of albumen of purifying, after the zymin of embodiment 1-3 gained and 4 × SDS sample-loading buffer being mixed with 1:3 volume ratio, be then splined on SDS-PAGE glue.After electrophoresis terminates under constant current 10 mA, unload lower glass plate, take out gel, after coomassie brilliant blue staining decolouring, take pictures with gel imaging instrument.The results are shown in accompanying drawing 1, wherein swimming lane M is Protein Marker; Swimming lane 1 is non-reduced state algin catenase; Swimming lane 2 is reduced state algin catenase (additionally adding 1% (V/V) beta-mercaptoethanol).Algin catenase molecular weight under non-reduced state is 28 kDa, is about 30 kDa under reduced state.
In order to verify that the albumen of embodiment 1-3 gained purifying is algin catenase, cut glue after being dyeed by the protein electrophoresis of embodiment 1 gained purifying to reclaim, by film dosim extracting enzymatic fragment, Matrix-assisted laser desorption ionization analysis is carried out after desalination, obtain peptide fingerprinting spectrum, peptide fingerprinting modal data will be obtained with Mascot software and carry out peptide section sequence comparison in NCBInr database.Comparison result is shown in accompanying drawing 2, as can be seen from the figure this algin catenase and European Bao spiral shell
haliotis tuberculatawith awabi's spiral shell
haliotis giganteahomology is respectively 92%, and 100%.Show that the albumen prepared is a kind of algin catenase.
In order to verify that the albumen of purifying is algin catenase, content is become to be the protein solution of 0.01 mg/mL the albumen of embodiment 1 gained purifying 20 mmol/L Tris-HCl (pH 8.5) buffer, get 1% (W/V) algin (Shanghai Chemical Reagent Co., Ltd., Sinopharm Group) that 1 mL joins 50 mL, 20 mmol/L Tris-HCl (pH 8.5) solution preparation, DNS method is used to measure product reducing sugar content, A
540be worth higher, reducing sugar content is higher.The results are shown in accompanying drawing 3, as seen from the figure with the prolongation in reaction times, reducing sugar content constantly increases, and proving that the albumen prepared has the ability of decomposing algin, is a kind of algin catenase.Meanwhile, this enzyme can be degraded algin effectively, reduces its viscosity, the results are shown in accompanying drawing 4, and as can be seen from Figure 4, along with time lengthening, viscosity reduces.
Although illustrate and describe embodiments of the invention above, be understandable that, above-described embodiment is exemplary, can not be interpreted as limitation of the present invention, those of ordinary skill in the art can change above-described embodiment within the scope of the invention when not departing from principle of the present invention and aim, revising, replacing and modification.
Claims (5)
1. with abalone internal organ for the method for algin catenase prepared by raw material, it is characterized in that comprising the steps:
Thick enzyme extraction: shredded by abalone internal organ, add Tris-HCl damping fluid, tissue mashing, centrifugal, supernatant liquor is crude enzyme liquid;
Activated carbon treatment: described crude enzyme liquid is crossed and filtered gac after activated carbon treatment, and gained is filtrate;
Anion exchange chromatography: by gained filtrate through DEAE-Sepharose or DEAE-Sephacel or Q-Sepharose anionite-exchange resin, the resin after absorption, through stepwise elution desorb, obtains the algin catenase of higher degree;
Film process 1: outer liquid is got in the ultra-filtration membrane ultrafiltration having algin catenase active part leacheate 100-50 kDa molecular weight to retain, obtains the algin catenase of purifying;
Film process 2: the ultra-filtration membrane ultrafiltration and concentration retained by outer for above-mentioned ultrafiltration liquid part 10-3 kDa molecular weight, obtains concentrated solution;
Enzyme preparation: by spray-dried for described concentrated solution or lyophilize, obtains the algin catenase goods of high vigor;
Optional, in described concentrated solution, add trehalose or beta-cyclodextrin before described spraying dry.
2. the preparation method of algin catenase described in claim 1, is characterized in that, in described thick enzyme extraction, adds the Tris-HCl damping fluid of 4 times of volume 20 mmol/L pH 8.5; Described centrifugal be centrifugal 20 min of 12000 × g.
3. the preparation method of algin catenase described in claim 1, is characterized in that, in described activated carbon treatment, described gac add-on is 0.5-3 bulking value %, and processing mode is 4 DEG C of stir process 30-60 min.
4. the preparation method of algin catenase described in claim 1, is characterized in that, in described anion exchange chromatography, described stepwise elution desorb be absorption after resin through 0.05,0.1,0.2,0.3 mol/L NaCl salt concn stepwise elution.
5. the preparation method of algin catenase described in claim 1, is characterized in that, in described enzyme preparation, the add-on of described trehalose or beta-cyclodextrin is 10-30 bulking value %.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510124968.XA CN104774827A (en) | 2015-03-23 | 2015-03-23 | Method for preparing alginate lyase from abalone internal organs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510124968.XA CN104774827A (en) | 2015-03-23 | 2015-03-23 | Method for preparing alginate lyase from abalone internal organs |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104774827A true CN104774827A (en) | 2015-07-15 |
Family
ID=53616598
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510124968.XA Pending CN104774827A (en) | 2015-03-23 | 2015-03-23 | Method for preparing alginate lyase from abalone internal organs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104774827A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105249434A (en) * | 2015-11-10 | 2016-01-20 | 集美大学 | Method for preparing natural alginate oligosaccharide water-retaining agent by utilizing abalone visceral enzyme liquid |
| CN105693031A (en) * | 2016-03-31 | 2016-06-22 | 北京建筑大学 | Method and device for synthesizing alginates in sewage treatment process |
| CN105996043A (en) * | 2016-05-31 | 2016-10-12 | 福建省恒祥渔业有限公司 | Oligo-uronic acid and polypeptide compound agent and preparation method thereof |
| CN112586696A (en) * | 2020-11-23 | 2021-04-02 | 集美大学 | Enzyme preparation for hydrolyzing laver and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103342668A (en) * | 2013-07-08 | 2013-10-09 | 集美大学 | Simple method for extracting natural taurine from abalone viscera |
| CN103589703A (en) * | 2013-11-26 | 2014-02-19 | 集美大学 | Method for preparing cellulase by using abalone viscera as raw materials |
| CN103805541A (en) * | 2014-01-21 | 2014-05-21 | 浙江树人大学 | Photobacterium phosphoreum and application thereof |
-
2015
- 2015-03-23 CN CN201510124968.XA patent/CN104774827A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103342668A (en) * | 2013-07-08 | 2013-10-09 | 集美大学 | Simple method for extracting natural taurine from abalone viscera |
| CN103589703A (en) * | 2013-11-26 | 2014-02-19 | 集美大学 | Method for preparing cellulase by using abalone viscera as raw materials |
| CN103805541A (en) * | 2014-01-21 | 2014-05-21 | 浙江树人大学 | Photobacterium phosphoreum and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 康平 等: ""皱纹盘鲍内脏酶的酶学性质及褐藻胶裂解酶的分离纯化"", 《水产学报》 * |
| 陶志鹏等: "皱纹盘鲍褐藻胶裂解酶的分离纯化及性质分析", 《科技与产业对接——CIFST-中国食品科学技术学会第十届年会暨第七届中美食品业高层论坛论文摘要集》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105249434A (en) * | 2015-11-10 | 2016-01-20 | 集美大学 | Method for preparing natural alginate oligosaccharide water-retaining agent by utilizing abalone visceral enzyme liquid |
| CN105693031A (en) * | 2016-03-31 | 2016-06-22 | 北京建筑大学 | Method and device for synthesizing alginates in sewage treatment process |
| CN105693031B (en) * | 2016-03-31 | 2018-09-18 | 北京建筑大学 | The method and equipment of alginates are synthesized in a kind of sewage disposal process |
| CN105996043A (en) * | 2016-05-31 | 2016-10-12 | 福建省恒祥渔业有限公司 | Oligo-uronic acid and polypeptide compound agent and preparation method thereof |
| CN105996043B (en) * | 2016-05-31 | 2019-07-09 | 福建省恒祥渔业有限公司 | A kind of oligomeric uronic acid and polypeptide complexing agent and preparation method thereof |
| CN112586696A (en) * | 2020-11-23 | 2021-04-02 | 集美大学 | Enzyme preparation for hydrolyzing laver and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101508690B (en) | Novel methods for simultaneously extracting tea polyphenol, tea polysaccharide and caffeinum from tea | |
| CN110623860B (en) | Giant salamander active peptide and hyaluronic acid composition capable of effectively promoting fibroblast proliferation | |
| CN102010460B (en) | Tea seed polypeptide and preparation method thereof | |
| CN101603038B (en) | Preparation method of lysozyme | |
| CN109651480A (en) | A method of separation momordica glycoside V | |
| CN101869169B (en) | Method for preparing fish oligopeptide from gurry by combining fermentation and membrane technology | |
| CN104774827A (en) | Method for preparing alginate lyase from abalone internal organs | |
| CN101029077B (en) | Method for purifying gene-recombinant insulin precursor | |
| CN107312807A (en) | The enzymolysis preparation of the brown alga function oligosaccharides in one main laminaria source | |
| CN104031158A (en) | Technique for extracting sulfate polysaccharide from Laminaria digitata | |
| WO2022067774A1 (en) | Preparation method and application of sea cucumber polysaccharide | |
| CN103073652A (en) | Method for extracting polysaccharide of spirulina platensis | |
| CN107586821A (en) | A kind of extracting method and purposes of saline cistanche polypeptide | |
| CN107011457B (en) | A method of extracting preparation non-starch polysaccharide and small molecule nutrient molecule from sweet potato waste water | |
| CN101570565A (en) | Functional component prepared from earthworms for promoting growth of microorganisms and improving culture units and preparation method thereof | |
| CN107721909B (en) | Method and system for continuously extracting DNJ, flavone and polysaccharide from Moraceae plant | |
| CN101845089B (en) | Method for large-scale production of neurotoxin in cobra venin and reduction of neurotoxicity | |
| CN103589703A (en) | Method for preparing cellulase by using abalone viscera as raw materials | |
| CN105166834A (en) | Preparation method of Maillard reaction products of ulva fasciata polysaccharide and gelatin peptide | |
| CN104087640A (en) | Cornua cervi pantotrichum polypeptide serial extraction method beneficial to skin or mucosa absorption | |
| CN102220296B (en) | Method for extracting garlic superoxide dismutase from garlic sheet processing waste water | |
| CN104055702B (en) | A kind of skin antiallergic wetting agent | |
| CN104758925A (en) | Iron chelation application of Trichiutus haumela bone iron-chelated collagen peptide | |
| CN1680220A (en) | Agent of extracting polyphenol from rape seed cakes or husks and its preparation | |
| CN102649823B (en) | Method for preparing copper polysaccharide through radial flow coupling ultra-filtering separation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150715 |
|
| RJ01 | Rejection of invention patent application after publication |