CN1206344C - Method for producing fucan sulfatase by means of bacteria - Google Patents
Method for producing fucan sulfatase by means of bacteria Download PDFInfo
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- CN1206344C CN1206344C CN 03138866 CN03138866A CN1206344C CN 1206344 C CN1206344 C CN 1206344C CN 03138866 CN03138866 CN 03138866 CN 03138866 A CN03138866 A CN 03138866A CN 1206344 C CN1206344 C CN 1206344C
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- enzyme
- bacterial strain
- producing
- fucoidan
- sulfatase
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- Enzymes And Modification Thereof (AREA)
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Abstract
The present invention relates to a method for producing fucosan sulfatase by bacteria. The method comprises the following steps: a culture medium containing an algae powder fermentation liquid is inoculated with a bacterial strain, and the bacterial strain is cultured at a certain temperature for a certain time; then an enzyme extract agent is put in the culture medium to make generated precipitates sink; separation is carried out; finally, drying by freezing is carried out. The method is characterized in that the bacterial strain used in the method is Vibrio fluvialis. The bacterial strain used in the method has the characteristics of wide range of nutritional requirements, easy culture and short generation time, particularly high enzyme producing activity; the enzyme activity of a fermentation liquid of the bacterial strain is 226 U/ml. The fucosan sulfatase produced by the method of the present invention has an interior contact type and an exterior contact type, and oligosaccharides, disaccharides and monosaccharides of the fucoidan can be obtained by degrading the fucoidan. The enzyme product has the advantages of high activity, high stability and low cost, and the industrialized production can be realized.
Description
Technical field
The present invention relates to a kind of method of producing the fucoidan enzyme with bacterium.
Background technology
Known to the inventor, the fucoidan enzyme has aspects such as the fucoidan oligosaccharide, disaccharide, monose of different molecular weight and demonstrates important role in preparation, it has decisive meaning to the development and the suitability for industrialized production of some marine drugs.At present, human Vibrio sp has been arranged, N-5 obtains circumscribed-type Fucose enzyme.Other has other microorganisms producing Fucose enzymes such as human Bacillus.But long with these bacterial classifications product enzyme times, the enzymic activity of output is low, cultivates complicated component, and difficulty is suitable for suitability for industrialized production.
Summary of the invention
The purpose of this invention is to provide a kind of method of producing the fucoidan enzyme with Vibrio flurialis, it can overcome the above-mentioned shortcoming of prior art.
A kind of method of producing the fucoidan enzyme with bacterium comprises bacterium is inserted in the seaweed powder fermented liquid, cultivates 18-24 hour under 18-28 ℃ culture temperature, drops into enzyme extracting agent (NH then
4)
2SO
4, the throw out of generation is sunk, separate again, last lyophilize is characterized in that used bacterial classification is Vibrio flurialis Vibrio fluvialis.
It is wide that the bacterial classification that the present invention uses has the nutritional requirement scope, cultivate easily and for the time short characteristics, particularly have high inulinase-producing activity, its fermentation broth enzyme vigor is 226U/ml.The fucoidan enzyme of producing with method of the present invention is inscribe and circumscribed two classes, and the degraded fucoidan can obtain fucoidan oligosaccharide, disaccharide, monose.The active height of this enzyme product, good stability, cost is low, can realize suitability for industrialized production.
Embodiment
The Vibrio flurialis that the present invention uses
Vibrio fluvialisBacterial strain is an arc, and end is given birth to single flagellum, G
-, bacterium colony circle, oyster white.The substratum of cultivating this bacterium employing is seawater bacteria substratum 2216E, and the component of substratum is yeast extract paste 1 gram, peptone 5 grams, and high ferric phosphate 0.01 gram, agar 15 grams, Chen Haishui 1000ml, its pH are 7.6-7.8.The used product enzyme substratum of the present invention is above-mentioned substratum, adds the seaweed powder of 0.8-1.5% (by weight, as follows) in addition again, cultivates and can obtain enzyme liquid in 20 hours.The depositary institution of above-mentioned Vibrio flurialis is Chinese typical culture collection center, is called for short CCTCC, and the address: Chinese Wuhan, Wuhan University, preservation date is on June 5th, 2000.Preserving number is M200015.
With Vibrio flurialis with the activation of above-mentioned 2216E substratum after, insert with 5% inoculum size and to be equipped with in the 500ml triangular flask that 100ml contains 1% algae powder substratum, behind 26 ℃ of shake-flask culture 18-24h, this nutrient solution is removed thalline with refrigerated centrifuge in centrifugal 10 minutes with 4000rpm, obtain fermenting enzyme liquid.This enzyme liquid is added (NH in 4 ℃ of refrigerators
4)
2SO
4Saturation ratio to 80%, 4 ℃ precipitate 24h down, the frozen centrifugation collecting precipitation, this precipitation is made crude zyme preparation after lyophilize.Described culture temperature is 18-28 ℃.
Claims (1)
1, a kind of method of producing the fucoidan enzyme with bacterium comprises bacterium is inserted in the seaweed powder fermented liquid, cultivates 18-24 hour under 18-28 ℃ culture temperature, drops into enzyme extracting agent (NH then
4)
2SO
4, the throw out of generation is sunk, separate again, last lyophilize is characterized in that used bacterial classification is Vibrio flurialis Vibrio fluvialis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03138866 CN1206344C (en) | 2003-07-28 | 2003-07-28 | Method for producing fucan sulfatase by means of bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03138866 CN1206344C (en) | 2003-07-28 | 2003-07-28 | Method for producing fucan sulfatase by means of bacteria |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1483814A CN1483814A (en) | 2004-03-24 |
| CN1206344C true CN1206344C (en) | 2005-06-15 |
Family
ID=34154934
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 03138866 Expired - Fee Related CN1206344C (en) | 2003-07-28 | 2003-07-28 | Method for producing fucan sulfatase by means of bacteria |
Country Status (1)
| Country | Link |
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| CN (1) | CN1206344C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107988109A (en) * | 2017-12-21 | 2018-05-04 | 青岛农业大学 | A kind of Flavobacterium mutant strain and its application |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103045512B (en) * | 2012-12-19 | 2014-07-16 | 青岛农业大学 | Flavobacterium and application thereof |
| CN103114063B (en) * | 2013-02-03 | 2014-03-19 | 中国海洋大学 | Strain for producing fucosan sulfatase and application thereof |
| IL312865A (en) | 2013-09-11 | 2024-07-01 | Eagle Biologics Inc | Liquid protein formulations containing viscosity-lowering agents |
| CN103834593B (en) * | 2014-03-05 | 2016-02-17 | 青岛农业大学 | A kind of secondary coccus and application thereof |
| CA2962768C (en) * | 2014-10-01 | 2023-10-10 | Alyssa M. Larson | Polysaccharide and nucleic acid formulations containing viscosity-lowering agents |
-
2003
- 2003-07-28 CN CN 03138866 patent/CN1206344C/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107988109A (en) * | 2017-12-21 | 2018-05-04 | 青岛农业大学 | A kind of Flavobacterium mutant strain and its application |
| CN107988109B (en) * | 2017-12-21 | 2019-12-10 | 青岛农业大学 | flavobacterium mutant strain and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1483814A (en) | 2004-03-24 |
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