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High-yield fermentation production process of Beta-agarase

A kind of agarase and process technology, applied in the direction of enzymes, enzymes, microorganisms, etc.

Inactive Publication Date: 2010-06-02
四川金稞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Purpose of the present invention: carry out ultraviolet mutagenesis breeding to Vibrio bacterium VB-3 with stronger agarase producing ability isolated from seawater, isolate and screen the agar with high enzyme activity, stable character and easy cultivation Agarase production strains, to solve the problem of key strains of fermentation production of agarase

Method used

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  • High-yield fermentation production process of Beta-agarase
  • High-yield fermentation production process of Beta-agarase
  • High-yield fermentation production process of Beta-agarase

Examples

Experimental program
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Embodiment 1

[0034] The screening of embodiment 1 agarase producing bacterial strain

[0035] The seawater collected from the ocean is dispersed in the culture medium dish that contains 1.5% agar, and the composition of culture medium is: peptone 5g, yeast extract 1g, ferric phosphate 0.1g, agar 15g, NaCl 20g, add water and be diluted to 1000ml. After culturing at 25°C, a transparent circle was formed around the agarase-producing colony, and the culture medium was concave in the shape of a kettle bottom. This feature was used as the basis for the initial screening of strains.

[0036] The strains screened were observed under a microscope, and the bacteria were curved and arc-shaped, with a single flagella; the experimental examination was Gram-negative bacteria, oxidase positive, catalase positive, 0 / 129 test positive, using glucose to produce acid. Gas production, according to the above characteristics, it was initially identified as Vibrio (Vibrio). The bacteria grow on the plate, can s...

Embodiment 2

[0038] Embodiment 2 Mutagenesis of agarase producing strain

[0039]First turn on the ultraviolet lamp (30W) in the inoculation room to preheat for 10 minutes, take 5ml of the prepared bacterial suspension and transfer it into a sterile petri dish with a diameter of 90mm, put a sterile magnetic stirring bar, and place it on a magnetic stirrer at a distance of 254nm from the wavelength. Open the lid of the dish at 30cm under the ultraviolet lamp and irradiate while stirring, and the doses are 15min respectively. After incubating the irradiated bacterial solution for 4 h, take 1 ml of the bacterial suspension with different irradiation time, and carry out 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 and 10 -7 Gradient dilution, take 10 -3 、10 -5 , and 10 -7 Spread 1ml of each concentration dilution on the separation plate, spread on the plate for primary screening, select colonies with fast growth and large transparent circles and transfer them to slant medium, culture at 2...

Embodiment 3

[0041] Seed culture and 5L fermenter cultivation of embodiment 3 mutant high-yield bacterial strains

[0042] 1. Seed culture

[0043] The composition of the seed medium: agar 2.0‰, yeast extract 0.3%, NH 4 Cl 1.0‰, K 2 HPO 4 0.1%, NaCl 1.0%, pH 8.0.

[0044] Temperature 25°C;

[0045] Shaker speed 150rpm;

[0046] Capacity 30ml / 250ml.

[0047] 2. Fermentation culture

[0048] The strains cultivated from the seeds were inoculated in a 5L fermenter for fermentation culture, the inoculation amount of the strains was 8%, and the amount of the fermentation medium was 2L.

[0049] The composition of the fermentation medium is: agar 2.0‰, yeast extract 0.1%, NH 4 Cl 1.0‰, K 2 HPO 4 0.12%, NaCl 1.0%, pH 8.0.

[0050] Fermentation temperature 25°C;

[0051] The initial pH value of fermentation is 8.0;

[0052] Fermentation stirring speed 150rpm;

[0053] The fermentation and cultivation time was 24h.

[0054] 3. Using a conventional method to separate and enrich the a...

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Abstract

The invention relates to a process condition for producing agarase by fermenting mutant breeding strains of vibrio which is a producing strain of Beta-agarase, and provides a fermentation culture process and an optional culture condition for breeding high-yield strain by mutagenesis. The invention determines the components of a slop culture medium, a seed culture medium, a fermentation culture medium and culture conditions, and the fermentation enzyme capability is more than 180U / ml; a conventional separation and purification method can be adopted to prepare the high-activity Beta-agarase. Therefore, the invention provides a production process of agarase, which has industrialized conversion prospects.

Description

technical field [0001] The invention relates to a process condition for fermenting and producing agarase by mutant Vibrio, mainly to determine the fermentation and culture conditions for high-yield agarase strains screened by an ultraviolet mutagenesis method and having industrialized production prospects. Background technique [0002] Agar (agar) is composed of agarose (agarose) and agaropectin (a-garopectin), and agarose is composed of (1→3)-O--βD-galactose and (1→4)-O-3,6 - Linear chain molecules composed of inner ether-α-L-galactose alternately; agar gel is composed of complex polysaccharide chains of galactose residues of different lengths, which contain various substituents such as sulfate groups, methyl groups, etc. , the exact details of its structure are still unclear. Natural agar mainly exists in the cell walls of some seaweeds. Agarase can hydrolyze agar polysaccharides to prepare agar oligosaccharides with a certain degree of polymerization and various physiolo...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12N15/01C12R1/63
Inventor 张春颖
Owner 四川金稞生物科技有限公司
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