Flavobacter johniiii, a strain producing alginate lyase

A technology of Flavobacterium johnsoni and Flavobacterium johniiii, applied in the biological field, can solve the problems of high price, difficult separation, low enzyme activity, etc., and achieve the effects of shortening the production cycle, reducing production costs, and simple nutritional requirements

Active Publication Date: 2020-02-11
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing alginate lyase generally has problems such as low enzyme activity, poor enzyme stability, tight combination of enzyme and product, and difficult separation. These shortcomings limit the popularization and application of alginate lyase.
Sigma company provides a kind of alginate lyase preparation product A1603, but the price is expensive, which greatly limits the wide application of alginate lyase and enzyme system in food, medicine, industry and agriculture, etc.

Method used

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  • Flavobacter johniiii, a strain producing alginate lyase
  • Flavobacter johniiii, a strain producing alginate lyase
  • Flavobacter johniiii, a strain producing alginate lyase

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Example 1: Isolation and identification of Flavobacterium johnsoniae (Flavobacterium johnsoniae) WX-11 and preservation of bacterial strains

[0021] (1) Isolation and screening of Flavobacterium johnsoniae WX-11

[0022] Dried kelp was purchased in the supermarket, cut into blocks, wrapped in gauze, and placed in conical flasks filled with river water, soil or tap water from the Jiangnan University campus. After incubation for 2 weeks, samples were taken. Shake the obtained sample in physiological saline with magnetic beads for 2 hours, let it stand overnight, inoculate it into a liquid medium with sodium alginate as the only carbon source, and culture it at 30°C and 220r / min until the blocky kelp is degraded and then transferred , five consecutive passages. The culture medium of continuous acclimatization for 5 generations was diluted with normal saline for 10 -7 、10 -8 、10 -9 times, spread on the solid medium with sodium alginate as the only carbon source on the ...

Embodiment 2

[0029] Embodiment 2: Preparation of alginate lyase by Flavobacterium johnsoniae WX-11

[0030] The method for producing alginate lyase by Flavobacter johnii WX-11 described in embodiment 1, the steps are as follows:

[0031] (1) Strain activation: inoculate Flavobacterium johnii WX-11 in a solid medium, and culture it for 24 hours at 25-30°C to obtain an activated strain;

[0032](2) Seed solution preparation: take the activated strain obtained in step (1), inoculate it in a 250mL Erlenmeyer flask containing 50mL of seed medium, and cultivate it for 8 ~12h, the seed liquid is obtained;

[0033] (3) Fermentation liquid preparation: get the seed liquid that step (2) makes, inoculate in the fermentation medium by volume ratio 1~5%, at 25~30 ℃, cultivate under the condition of 150~220r / min 18~ After 20 hours, the cell fermentation broth was obtained.

[0034] The formulations of seed medium and fermentation medium are: sodium alginate 1-5g / L, peptone 1-5g / L, yeast powder 1-5g / L...

Embodiment 3

[0039] Example 3: Analysis of degradation products of alginate lyase produced by Flavobacterium johnsoniae (Flavobacterium johnsoniae) WX-11

[0040] Take 1 g of sodium alginate in 100 mL of 50 mM (pH 7.0) phosphate buffer, add the crude alginate lyase solution produced by Flavobacter johnii WX-11, enzymolyze it in a water bath at 40 °C for 24 hours, and pass thin-layer chromatography Analysis of degradation products. The results show( figure 2 ), the product after enzymatic hydrolysis of sodium alginate is mainly 2-6 sugars, no monosaccharides are detected, it is speculated that it is an endo-type alginate lyase.

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Abstract

The invention discloses a strain of Flavobacter johniii producing alginate lyase, which belongs to the field of biotechnology. The present invention isolates Flavobacterium johnsoniae WX-11 from soil and river water samples, that is, Flavobacterium johnsoniae CGMCC No.14444. The strain has simple nutritional requirements and short fermentation time, and is used in the production of alginate lyase The production cycle can be significantly shortened and the production cost can be reduced. The invention also discloses the use of the strain to prepare alginate lyase and reduce the viscosity of high-viscosity raw materials containing alginate. The alginate lyase produced by the Flavobacterium johniiii WX-11 provided by the present invention can degrade sodium alginate to generate biologically active fucoidan oligosaccharides, which can be widely used in the fields of agriculture, food, feed addition, medicine and seaweed processing, etc. .

Description

technical field [0001] The invention relates to a strain of Flavobacter johniii producing alginate lyase, which belongs to the field of biotechnology. Background technique [0002] my country is rich in marine resources and is the country with the largest scale of marine algae cultivation, processing and utilization in the world. Seaweed production accounts for about 60% of the world's total output, exceeding 13 million tons. This rich algae contains a large amount of polysaccharides such as alginate and sugar alcohols, making seaweed processing and its high-value utilization an important field of comprehensive utilization of marine resources in my country. [0003] Studies have found that alginate degradation products, alginate oligosaccharides, have various biological activities, including growth-promoting effects, enhancing immunity, anti-tumor activity, neuroprotective effects, etc. Therefore, the development of functional oligosaccharides in alginate degradation product...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/88C12P19/12C12P19/00C12R1/20
CPCC12N1/20C12N9/88C12P19/00C12P19/12C12Y402/02C12N1/205C12R2001/20
Inventor 李恒史劲松郝瑶许正宏龚劲松李会季珂杨敏
Owner JIANGNAN UNIV
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