CN117265042A - Extraction process of land-based chitin - Google Patents
Extraction process of land-based chitin Download PDFInfo
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- CN117265042A CN117265042A CN202311066997.6A CN202311066997A CN117265042A CN 117265042 A CN117265042 A CN 117265042A CN 202311066997 A CN202311066997 A CN 202311066997A CN 117265042 A CN117265042 A CN 117265042A
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- 229920002101 Chitin Polymers 0.000 title claims abstract description 35
- 238000000605 extraction Methods 0.000 title claims abstract description 10
- 238000002360 preparation method Methods 0.000 claims abstract description 25
- 238000006243 chemical reaction Methods 0.000 claims abstract description 22
- 108090000790 Enzymes Proteins 0.000 claims abstract description 17
- 102000004190 Enzymes Human genes 0.000 claims abstract description 17
- 229940088598 enzyme Drugs 0.000 claims abstract description 17
- 239000012535 impurity Substances 0.000 claims abstract description 11
- 108010001682 Dextranase Proteins 0.000 claims abstract description 8
- 108010022172 Chitinases Proteins 0.000 claims abstract description 4
- 102000012286 Chitinases Human genes 0.000 claims abstract description 4
- 108010015776 Glucose oxidase Proteins 0.000 claims abstract description 4
- 239000004366 Glucose oxidase Substances 0.000 claims abstract description 4
- 238000004140 cleaning Methods 0.000 claims abstract description 4
- 230000000593 degrading effect Effects 0.000 claims abstract description 4
- 239000003344 environmental pollutant Substances 0.000 claims abstract description 4
- 229940116332 glucose oxidase Drugs 0.000 claims abstract description 4
- 235000019420 glucose oxidase Nutrition 0.000 claims abstract description 4
- 238000000227 grinding Methods 0.000 claims abstract description 4
- 231100000719 pollutant Toxicity 0.000 claims abstract description 4
- 241000238631 Hexapoda Species 0.000 claims abstract description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 48
- 235000006408 oxalic acid Nutrition 0.000 claims description 16
- 239000000243 solution Substances 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 230000008859 change Effects 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 238000011010 flushing procedure Methods 0.000 claims description 6
- 230000035484 reaction time Effects 0.000 claims description 6
- 238000001291 vacuum drying Methods 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 4
- 238000000746 purification Methods 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 238000010306 acid treatment Methods 0.000 claims description 3
- 239000007853 buffer solution Substances 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- 239000007795 chemical reaction product Substances 0.000 claims description 3
- 230000018044 dehydration Effects 0.000 claims description 3
- 238000006297 dehydration reaction Methods 0.000 claims description 3
- 230000002255 enzymatic effect Effects 0.000 claims description 3
- 238000006911 enzymatic reaction Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 239000000413 hydrolysate Substances 0.000 claims description 3
- 238000004255 ion exchange chromatography Methods 0.000 claims description 3
- 238000001556 precipitation Methods 0.000 claims description 3
- 239000000047 product Substances 0.000 claims description 3
- 238000002791 soaking Methods 0.000 claims description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 238000000108 ultra-filtration Methods 0.000 claims description 3
- 238000009423 ventilation Methods 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 2
- 229920001661 Chitosan Polymers 0.000 description 3
- 241000238557 Decapoda Species 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 241000238424 Crustacea Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 241000237536 Mytilus edulis Species 0.000 description 1
- 241000382353 Pupa Species 0.000 description 1
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000010794 food waste Substances 0.000 description 1
- 229960002442 glucosamine Drugs 0.000 description 1
- MSWZFWKMSRAUBD-IVMDWMLBSA-N glucosamine group Chemical group OC1[C@H](N)[C@@H](O)[C@H](O)[C@H](O1)CO MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000020638 mussel Nutrition 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Classifications
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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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
- C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
-
- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Materials Engineering (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses an extraction process of land-based chitin, which comprises the following steps: step one, sample preparation: collecting a fly maggot sample from a culture environment of fly maggots, wherein the fly maggots are larva stages of insects, cleaning the collected fly maggot sample, removing impurities and pollutants, and grinding or crushing the fly maggot sample to increase the surface area of the sample; step two, enzyme preparation selection: selecting an enzyme preparation suitable for degrading chitin, wherein one or more of chitinase, dextranase, beta-1, 3-dextranase, beta-N-acetamido dextranase and glucose oxidase are selected as the enzyme preparation; step three, enzymolysis reaction: placing the crushed maggot sample and the selected enzyme preparation in the same reaction container, carrying out mixed reaction under the proper temperature, pH value and time conditions, determining the enzymolysis time according to the experimental requirements and sample characteristics, and controlling the enzymolysis time to be 5-10 h. The invention can effectively extract the chitin, meets the use requirement of the chitin and ensures the use effect.
Description
Technical Field
The invention relates to the technical field of chitin preparation, in particular to an extraction process of land-based chitin.
Background
Chitin is a major component present in crustacean shells and shell organisms and is a complex polysaccharide polymer. Chitin consists of two main polysaccharides: chitosan and chitin acid. Chitosan is a main component of chitin, and its structure is composed of glucosamine units, is a linear polymer, and is formed by connecting beta-1, 4-glucosamine bonds. Chitin acid is formed by partial removal of acetyl groups on chitosan.
Chitin, which is widely present in nature in the exoskeletons of crustaceans (e.g., shrimps, crabs, lobsters, etc.) and shells of shell-like organisms (e.g., mussels, etc.), is an important component of the protection and support of body structures by these organisms, and has a certain mechanical strength and biodegradability, and thus has a wide range of applications in many fields, such as agriculture, foods, medicine, environment, etc.
The fly maggots are fly larvae, the growth and propagation are extremely fast, a lot of equipment is not needed for artificial cultivation, the indoor and outdoor cultivation and urban rural cultivation can be realized, the high-technology sterile fly maggots can be conditionally propagated for comprehensive development, and the chitin can be extracted from the surfaces and the pupa shells of the fly maggots, so that the extraction process of the land-based chitin is provided.
Disclosure of Invention
The invention aims to provide an extraction process of land-based chitin, which aims to solve the problems in the background technology.
In order to achieve the above purpose, the present invention provides the following technical solutions: the method comprises the following steps:
step one, sample preparation: collecting a fly maggot sample from a culture environment of fly maggots, wherein the fly maggots are larva stages of insects, cleaning the collected fly maggot sample, removing impurities and pollutants, and grinding or crushing the fly maggot sample to increase the surface area of the sample;
step two, enzyme preparation selection: selecting an enzyme preparation suitable for degrading chitin, wherein one or more of chitinase, dextranase, beta-1, 3-dextranase, beta-N-acetamido dextranase and glucose oxidase are selected as the enzyme preparation;
step three, enzymolysis reaction: placing the crushed maggot sample and the selected enzyme preparation in the same reaction container, carrying out mixed reaction under the proper temperature, pH value and time conditions, determining enzymolysis time according to experimental requirements and sample characteristics, and controlling the enzymolysis time to be 5-10 h;
step four, separation and purification: separating the enzymatic hydrolysate from the residue by filtration, centrifugation, precipitation, etc., and further purifying by ultrafiltration, ion exchange chromatography, etc., to obtain pure chitin extract;
step five, drying and preserving: drying the chitin extract by low temperature or vacuum drying, and storing in a dry and sealed container to maintain stability and quality.
Preferably, the specific steps of sample preparation in the step one are as follows:
(1) Removing solid impurities: firstly, placing a fly maggot sample on a filter screen or a screen, and flushing the sample with flowing water;
(2) Surface disinfection: immersing the cleaned maggot sample in a container containing 70% ethanol solution, and gently stirring or shaking for a period of time;
(3) Oxalic acid treatment: preparing 10% oxalic acid solution, soaking a maggot sample in the oxalic acid solution, and keeping for 6-10 minutes;
(4) And (5) flushing again: thoroughly washing the fly maggot sample in the oxalic acid solution by using flowing clear water to remove residual oxalic acid and impurities;
(5) And (3) placing the treated maggot sample in a ventilation place or using a centrifuge to carry out centrifugal dehydration so as to remove redundant moisture.
Preferably, the temperature control in the third step can be performed by placing the reaction vessel in water at a desired temperature using a water bath or a thermostat or a device with a temperature sensor and a control function, and the control of the pH value can be performed by adding a suitable buffer solution to the reaction system to resist the change of the pH value during the enzyme reaction and combining the device with the pH sensor and the control function.
Preferably, in the third step, different reaction time points can be tried, and the reaction product at each time point is detected and analyzed, so that the optimal enzymolysis reaction time can be determined by observing the trend of the product generation and the change of the reaction rate.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the invention, the enzyme preparation is used for carrying out enzymolysis reaction on the treated fly maggot sample, and then the purified chitin extract is obtained through separation and purification, so that the chitin can be effectively extracted, and the use requirement of the chitin is met;
2. the invention also dries the chitin by low temperature or vacuum drying, and stores the extracted chitin in a dry and sealed container to keep the stability and quality and ensure the use effect.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The invention provides a technical scheme that: the method comprises the following steps:
step one, sample preparation: collecting a maggot sample from a maggot culture environment, cleaning the collected maggot sample, removing impurities and pollutants, and grinding or crushing the maggot sample to increase the surface area of the sample;
step two, enzyme preparation selection: selecting an enzyme preparation suitable for degrading chitin, wherein one or more of chitinase, dextranase, beta-1, 3-dextranase, beta-N-acetamido dextranase and glucose oxidase are selected as the enzyme preparation;
step three, enzymolysis reaction: placing the crushed maggot sample and the selected enzyme preparation in the same reaction container, carrying out mixed reaction under the proper temperature, pH value and time conditions, determining enzymolysis time according to experimental requirements and sample characteristics, and controlling the enzymolysis time to be 5-10 h;
step four, separation and purification: separating the enzymatic hydrolysate from the residue by filtration, centrifugation, precipitation, etc., and further purifying by ultrafiltration, ion exchange chromatography, etc., to obtain pure chitin extract;
step five, drying and preserving: drying the chitin extract by low temperature or vacuum drying, and storing in a dry and sealed container to maintain stability and quality.
The sample preparation in the first step comprises the following specific steps:
(1) Removing solid impurities: firstly, placing a fly maggot sample on a filter screen or a screen, and flushing the sample with flowing water, so that most solid impurities such as food residues, dirt and the like can be removed;
(2) Surface disinfection: immersing the cleaned maggot sample in a container containing 70% ethanol solution, and gently stirring or shaking for a period of time, which is effective in eliminating bacteria and pathogens on the surface; this can effectively eliminate bacteria and pathogens on the surface;
(3) Oxalic acid treatment: preparing 10% oxalic acid solution, soaking a maggot sample in the oxalic acid solution, and keeping for 6-10 minutes; oxalic acid can help to remove organic matters on the fly maggot exoskeleton and help to kill microorganisms which may exist;
(4) And (5) flushing again: thoroughly washing the fly maggot sample in the oxalic acid solution with flowing clear water to remove residual oxalic acid and impurities, and repeatedly washing for several times until the solution becomes clear and has no color;
(5) And (3) placing the treated maggot sample in a ventilation place or using a centrifuge for centrifugal dehydration to remove redundant moisture, and preferably using a low-temperature or vacuum drying method to completely dry the sample.
The temperature control in the third step can be realized by placing the reaction vessel in water with the required temperature or equipment with temperature sensor and control function, such as a constant temperature incubator, a thermal cycler and the like, controlling the temperature to 30-50 ℃, adding proper buffer solution into the reaction system to resist the change of the pH value in the enzyme reaction process, and combining the equipment with pH sensor and control function, such as a pH meter and a pH regulator, controlling the pH value to be 6.0-8.0.
In the third step, different reaction time points can be tried, and the reaction products at each time point can be detected and analyzed, so that the optimal enzymolysis reaction time can be determined by observing the trend of the product generation and the change of the reaction rate.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (4)
1. The extraction process of the land-based chitin is characterized by comprising the following steps of:
step one, sample preparation: collecting a fly maggot sample from a culture environment of fly maggots, wherein the fly maggots are larva stages of insects, cleaning the collected fly maggot sample, removing impurities and pollutants, and grinding or crushing the fly maggot sample to increase the surface area of the sample;
step two, enzyme preparation selection: selecting an enzyme preparation suitable for degrading chitin, wherein one or more of chitinase, dextranase, beta-1, 3-dextranase, beta-N-acetamido dextranase and glucose oxidase are selected as the enzyme preparation;
step three, enzymolysis reaction: placing the crushed maggot sample and the selected enzyme preparation in the same reaction container, carrying out mixed reaction under the proper temperature, pH value and time conditions, determining enzymolysis time according to experimental requirements and sample characteristics, and controlling the enzymolysis time to be 5-10 h;
step four, separation and purification: separating the enzymatic hydrolysate from the residue by filtration, centrifugation, precipitation, etc., and further purifying by ultrafiltration, ion exchange chromatography, etc., to obtain pure chitin extract;
step five, drying and preserving: drying the chitin extract by low temperature or vacuum drying, and storing in a dry and sealed container to maintain stability and quality.
2. The extraction process of land-based chitin according to claim 1, wherein: the sample preparation in the first step comprises the following specific steps:
(1) Removing solid impurities: firstly, placing a fly maggot sample on a filter screen or a screen, and flushing the sample with flowing water;
(2) Surface disinfection: immersing the cleaned maggot sample in a container containing 70% ethanol solution, and gently stirring or shaking for a period of time;
(3) Oxalic acid treatment: preparing 10% oxalic acid solution, soaking a maggot sample in the oxalic acid solution, and keeping for 6-10 minutes;
(4) And (5) flushing again: thoroughly washing the fly maggot sample in the oxalic acid solution by using flowing clear water to remove residual oxalic acid and impurities;
(5) And (3) placing the treated maggot sample in a ventilation place or using a centrifuge to carry out centrifugal dehydration so as to remove redundant moisture.
3. The extraction process of land-based chitin according to claim 1, wherein: the temperature control in the third step can be realized by using a water bath or a thermostat to place the reaction vessel in water with the required temperature or equipment with a temperature sensor and a control function, and the control of the PH value can be realized by adding proper buffer solution into a reaction system to resist the change of the PH value in the enzyme reaction process and combining the equipment with the pH sensor and the control function.
4. The extraction process of land-based chitin according to claim 1, wherein: in the third step, different reaction time points can be tried, and the reaction products at each time point can be detected and analyzed, so that the optimal enzymolysis reaction time can be determined by observing the trend of the product generation and the change of the reaction rate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311066997.6A CN117265042A (en) | 2023-08-23 | 2023-08-23 | Extraction process of land-based chitin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311066997.6A CN117265042A (en) | 2023-08-23 | 2023-08-23 | Extraction process of land-based chitin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117265042A true CN117265042A (en) | 2023-12-22 |
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ID=89220447
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311066997.6A Pending CN117265042A (en) | 2023-08-23 | 2023-08-23 | Extraction process of land-based chitin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117265042A (en) |
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2023
- 2023-08-23 CN CN202311066997.6A patent/CN117265042A/en active Pending
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