CN103642775A - Chitinase preparation method - Google Patents
Chitinase preparation method Download PDFInfo
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- CN103642775A CN103642775A CN201310588354.8A CN201310588354A CN103642775A CN 103642775 A CN103642775 A CN 103642775A CN 201310588354 A CN201310588354 A CN 201310588354A CN 103642775 A CN103642775 A CN 103642775A
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- Prior art keywords
- chitinase
- enzyme
- preparation
- carrying
- fermentation
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- 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
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
- C12N9/2442—Chitinase (3.2.1.14)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01014—Chitinase (3.2.1.14)
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- 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)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention discloses a chitinase preparation method, which specifically comprises: carrying out complete culture on target bacteria, wherein low temperature culture is performed from the beginning stage of the growth period until the final stage of the growth period is achieved, and then the obtained target bacterial is taken out and placed in a centrifuge tube; adding a fermentation enzyme and cellulose with a mass fraction of 1-2%; filtering with five layers of gauzes, and taking the precipitate; adding a sodium alginate solution with a mass fraction of 5%, and completely stirring and mixing to obtain a glass bead-like ball; carrying out hot bath for 15 min in water with a temperature of 85 DEG C, and adding a potassium cyanide solution; and carrying out high speed centrifugation, carrying out oscillation, and taking out, wherein the fermentation period is 35 h, and the target bacterial is streptomycete. The chitinase preparation method has the following beneficial effects that: the high yield required enzyme can be obtained at various stages, the enzyme fermentation activity is high, the enzyme can be used in actual production, and characteristics of small particle and homology are provided.
Description
Technical field
The present invention relates to the preparation method of biochemical substances, specifically, refer to a kind of preparation method of chitinase.
Background technology
Chitin has another name called chitosan, is the chitin in the crust in Crustacean (as shrimp, crab), insect and other invertebrates shells, and a kind of natural macromolecule amylose body making through deacetylated (extraction), is the foodstuff fibre of animality.Chitin is de-
secondthe degree of acidylate is higher, and the physiological effect of performance is also stronger, and still conspicuous chitin deacetylation can reach more than 90%.International Medical nutritive food association will
thiskind of material called after except sugar, protein, fat and, VITAMIN and mineral substance students from five types of universities order the sixth-largest vital principle after key element, so are more and more subject to extensive concern, it is different from general natural nutriment completely, it is exactly chitin.The hydrolysis of catalysis chitin generates the enzyme of N-acetyl-glucosamine reaction.EC?3．2．1．14。Be that Ka Le and Huffman (P.Karrer, A.Hofmann, 1929) are found from the gastric juice (mid intestinal gland juice) of snail (Helix pomatia), character is not quite clear.In addition, in the exuvial gland juice of insect, also can find, there is digestion and absorb old cuticular effect.In the moulting fluid of insect, the pericarp of almond (almond) and mustiness bacterium, also find to some extent.
Summary of the invention
For overcoming above-mentioned technical problem, we have proposed following technical scheme:
A preparation method for chitinase, concrete steps are as follows:
First object bacterium is cultivated completely, required to start to carry out low temperature cultivation from its early growth period, then to it, grow latter stage, take out and be positioned in centrifuge tube;
Add fermenting enzyme, and massfraction reaches percent 1 to percent 2 Mierocrystalline cellulose;
Five layers of gauze filter, and get its throw out;
Adding massfraction is 5 percent sodium alginate soln, is fully uniformly mixed, and obtains glass pearl ball;
In the water of 85 degrees Celsius, carry out heating bath 15 minutes, then add potassium cyanide solution;
High speed centrifugation, vibration, takes out.
In the present invention, described fermentation period is 35 hours.
In the present invention, described object bacterium is streptomycete.
The invention has the beneficial effects as follows: can all obtain in each period the required enzyme of high yield, flushing of enzymic fermentation, can in actual production activity, apply, and particle is smaller, has homology.
Embodiment
A preparation method for chitinase, concrete steps are as follows:
First object bacterium is cultivated completely, required to start to carry out low temperature cultivation from its early growth period, then to it, grow latter stage, take out and be positioned in centrifuge tube;
Add fermenting enzyme, and massfraction reaches percent 1 Mierocrystalline cellulose;
Five layers of gauze filter, and get its throw out;
Adding massfraction is 5 percent sodium alginate soln, is fully uniformly mixed, and obtains glass pearl ball;
In the water of 85 degrees Celsius, carry out heating bath 15 minutes, then add potassium cyanide solution;
High speed centrifugation, vibration, takes out.
Described fermentation period is 35 hours.
Described object bacterium is streptomycete.
The above; it is only preferably embodiment of the present invention; but protection scope of the present invention is not limited to this; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses; according to technical scheme of the present invention and inventive concept thereof, be equal to replacement or changed, within all should being encompassed in protection scope of the present invention.
Claims (3)
1. a preparation method for chitinase, is characterized in that, concrete steps are as follows:
First object bacterium is cultivated completely, required to start to carry out low temperature cultivation from its early growth period, then to it, grow latter stage, take out and be positioned in centrifuge tube;
Add fermenting enzyme, and massfraction reaches percent 1 to percent 2 Mierocrystalline cellulose;
Five layers of gauze filter, and get its throw out;
Adding massfraction is 5 percent sodium alginate soln, is fully uniformly mixed, and obtains glass pearl ball;
In the water of 85 degrees Celsius, carry out heating bath 15 minutes, then add potassium cyanide solution;
High speed centrifugation, vibration, takes out.
2. the preparation method of a kind of chitinase as claimed in claim 1, is characterized in that, described fermentation period is 35 hours.
3. the preparation method of a kind of chitinase as claimed in claim 1, is characterized in that, described object bacterium is streptomycete.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310588354.8A CN103642775A (en) | 2013-11-21 | 2013-11-21 | Chitinase preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310588354.8A CN103642775A (en) | 2013-11-21 | 2013-11-21 | Chitinase preparation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103642775A true CN103642775A (en) | 2014-03-19 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310588354.8A Pending CN103642775A (en) | 2013-11-21 | 2013-11-21 | Chitinase preparation method |
Country Status (1)
| Country | Link |
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| CN (1) | CN103642775A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109337843A (en) * | 2018-11-19 | 2019-02-15 | 常熟理工学院 | One plant of production chitinase bacterial strain and application |
-
2013
- 2013-11-21 CN CN201310588354.8A patent/CN103642775A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109337843A (en) * | 2018-11-19 | 2019-02-15 | 常熟理工学院 | One plant of production chitinase bacterial strain and application |
| CN109337843B (en) * | 2018-11-19 | 2020-12-15 | 常熟理工学院 | Bacterial strain for producing chitinase and application thereof |
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| PB01 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140319 |