A kind of endo-algin lyase, its encoding gene and application

A technology of endo-algin cutting and cutting algin, which is applied in the field of bioengineering, can solve the problems of industrial application limitations, interference with alginate oligosaccharide correlation analysis, poor acid-base and metal ion tolerance, etc.
CN113046378BActive Publication Date: 2022-08-05THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES
Publication Date
2022-08-05

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Abstract

The invention discloses an endo-algin lyase, its encoding gene and application. The present invention takes the genomic DNA of Vibrio sp.MCCC 1A13243 as a template, designs and synthesizes primers, amplifies the endo-alginate lyase gene by PCR, and clones the gene into the pET-22b expression vector. Induced expression in BL21 and purified by Ni-Sepharose affinity chromatography to obtain a higher-purity recombinant endo-algin lyase protein. The optimum temperature of the enzyme is 35℃, the optimum pH is 8.5, and the stability is good in the range of 0~65℃ and pH 4.5~10.5; Co 2+ , Cu 2+ , Mn 2+ and Fe 3+ It can obviously promote the activity of the enzyme; the enzyme has the strongest degradation effect on sodium alginate, and can also degrade polyguluronic acid and polymannuronic acid, and the main product of degrading these three substrates is disaccharide. The enzyme has advantages in the production of fucoidan oligosaccharides, especially fucoidobiose.
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Description

technical field

[0001] The invention relates to obtaining an endo-alginate lyase by means of genetic engineering and a preparation method thereof, and belongs to the technical field of bioengineering. Background technique

[0002] Algin is a kind of water-soluble acidic polysaccharide mainly extracted from the cell wall of brown algae. It is randomly arranged by two monomers, β-D-mannuronic acid (M) and α-L-guluronic acid (G). composed of polymers, the two monomers are linked by 1,4-glycosidic bonds to form three different forms of polysaccharide fragments: poly-β-D-mannuronic acid (Polymannuronic acid, PolyM) fragment, polyα-L -Guluronic acid (Polyguluronic acid, PolyG) fragment and both hybrid fragments (PolyMG). Because of its physiological activities such as anti-tumor and lowering blood pressure, algin is widely used in food, medical treatment, agriculture, energy and so on.

[0003] At present, biological enzymatic method is an important method for degrading algin. C...

Claims

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