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High-yield fucoidanase ocean alternating pseudomonas and application thereof

A technology of Pseudomonas alternata and fucoidan, applied in the direction of bacteria, hydrolase, and microbial-based methods, can solve the problems of low activity and unrealized commercial production, and achieve short fermentation cycle and high enzyme activity Effect

Active Publication Date: 2017-05-10
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the low activity of this type of enzyme, the commercial production of this enzyme has not been realized, so screening high-activity enzyme-producing microbial strains is still an important issue

Method used

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  • High-yield fucoidanase ocean alternating pseudomonas and application thereof
  • High-yield fucoidanase ocean alternating pseudomonas and application thereof
  • High-yield fucoidanase ocean alternating pseudomonas and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: Screening and identification of microorganisms

[0017] 1. Screening of microorganisms

[0018] Live scallops, sea urchins and abalones are dissected under sterile conditions, and their digestive glands are separated, and sterile saline is added to the extracted marine biological digestive glands and rotten kelp, and the digestive glands of scallops, sea urchins and abalones are obtained after extraction And the tissue suspension of rotten kelp. Take 1-2 mL of the tissue suspension and inoculate it into fucoidan sulfate liquid selection medium A, and culture in a shaking incubator at 25°C and 150 rpm. After 24 hours, the fucoidan-producing enzyme activity of each strain was measured, and 100-200 μL of the bacterial solution with enzyme activity was selected and spread on the fucoidan sulfate solid selective medium B. Place the culture plate in a 25°C incubator for cultivation. Pick colonies with good growth status from the plate and inoculate them in fu...

Embodiment 2

[0030] Embodiment 2: the fermentative production enzyme of microorganism

[0031] 1. Fermentation culture

[0032] The highly active bacterial strain obtained by screening is fermented and cultured, and the growth curve of the microorganism in the fucoidan sulfate medium is obtained.

[0033] According to the inoculum amount of 1-3%, insert the seed solution into the fucoidan sulfate liquid fermentation medium, cultivate it in a shaker at 25-37°C and 150rpm, collect the fermentation solution at different times, and use the uninoculated culture medium As a blank control, the absorbance value at 600 nm of the bacterial solution was measured, and the growth curve of the bacteria was drawn ( figure 1 ). As a result, it was found that the bacteria reached the plateau stage of growth in about 18-24 hours in the fucoidan sulfate medium, and at this time the OD 600 = about 5.3. This is the best time to collect bacteria in the future.

[0034] 2. Extraction and activity determinat...

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PUM

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Abstract

The invention provides a high-yield fucoidanase degrading enzyme strain. The preservation number of the strain is CGMCC No.13477. The microbiological strain is used for producing fucoidanase, the fucoidanase produced by the screened strain can degrade fucoidan, and oligomerization fucoidanase with potential biological activity is generated. The microorganism has the advantages that the fermentation period is short, the enzyme activity is high, and most activity is intracellular activity, collection and preservation of samples are promoted, and a foundation is laid for subsequent industrial application.

Description

technical field [0001] The invention belongs to the technical field of marine microorganism screening, and in particular relates to a marine alternate pseudomonas with high fucoidanase production and application thereof. Background technique [0002] Fucoidan is a water-soluble polysaccharide contained in the cell wall of brown algae, mainly composed of L-fucose and different sulfated L-fucose. In addition, it also contains a certain amount of D-galactose, D-mannose, D-glucuronic acid, a small amount of D-xylose and L-arabinose, which is a high molecular weight acidic acid with very complex chemical composition and structure. Polysaccharide compounds. Studies have found that this type of polysaccharide has various biological activities such as anti-oxidation, lowering blood fat, anti-coagulation, anti-virus, anti-tumor, enhancing body immunity, etc. important relationship. Therefore, in order to study the structure-activity relationship of this type of polysaccharide in d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/24C12R1/38
CPCC12N1/20C12N9/24C12N1/205C12R2001/38
Inventor 刘伟治于广利张柯柯律倩倩赵小亮
Owner OCEAN UNIV OF CHINA
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