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Beta-glucanase generated by Bacillus marinus and preparation method thereof

A technology of marine bacillus and glucanase, which is applied in the field of beta-glucanase produced by marine bacillus and its preparation, can solve the problems of waste of resources, environmental pollution, difficult production scale, complicated operation, etc. Application prospect, easy storage, high purity effect

Inactive Publication Date: 2016-10-12
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But acid hydrolysis, chemical synthesis and other processes are complex in operation, high in energy consumption, and low in yield, making it difficult to realize large-scale production; and using β-glucanase to hydrolyze wheat straw to produce functional oligosaccharides, the operation process is simple and can be greatly improved. Reduce production costs, improve the utilization rate of wheat straw, reduce the waste of biomass resources, and reduce the air pollution caused by straw burning, which has great social, economic and environmental value
Saccharomyces cerevisiae autolyzed residues containing a large amount of β-1,3-glucan are mostly used as industrial waste or sold as feed at a low price, resulting in a great waste of resources and environmental pollution

Method used

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  • Beta-glucanase generated by Bacillus marinus and preparation method thereof
  • Beta-glucanase generated by Bacillus marinus and preparation method thereof
  • Beta-glucanase generated by Bacillus marinus and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Morphological characteristics, physiological and biochemical characteristics and 16S rDNA sequence analysis of strain N6-2 of embodiment 1

[0039] (1) Morphological characteristics

[0040] The results of Gram staining showed that the strain N6-2 was a Gram-negative bacterium with obvious spores, and the shape of the bacteria was short rods and rods, and most of them were connected by two bacteria. The colonies are characterized by white or milky-yellow white, with smooth surface and protrusions. Scanning electron microscope pictures show that the strain N6-2 is in the shape of a short rod, the surface of the bacteria is smooth, and the two ends show protrusions. The middle is sunken, the connection is slender, and there is mucous membrane between the bacteria. The size of a single bacterium is width: 0.9-1.5 μm, length: 1-3.7 μm.

[0041] (2) Physiological and biochemical reaction characteristics

[0042] Because the bacterium is Gram-negative bacteria, it can use...

Embodiment 2

[0054] Example 2 Separation and purification of β-glucanase produced by marine bacillus N6-2

[0055] (1) Fermentation of bacterial strain:

[0056] Get marine bacillus N6-2 bacterial strain, inoculate in the test tube that contains seed culture medium, in the shaker of 28 ℃, 150rpm, culture 28h; The bacterial strain after above-mentioned culture is inoculated by 4% (v / v) inoculum amount in The fermentation medium was cultured on a shaker at 28°C and 150rpm for 80 hours; the fermentation liquid of the strain was taken, centrifuged, the bacteria and impurities in the fermentation liquid were removed, and the pH was adjusted to 7.0, which was the crude fermentation sample.

[0057] (2) Ultrafiltration interception:

[0058] Take the marine bacillus N6-2 fermentation broth, centrifuge at 8000rpm, remove the bacteria and some impurities. Use filter membranes with molecular weights of 50kDa, 30kDa, 10kDa, 5kDa, and 3kDa in turn to intercept the fermentation broth, divide the ferm...

Embodiment 3

[0066] Example 3 Detection of the purity of the β-glucanase produced by Bacillus marine bacillus N6-2

[0067] 1. Protein-PAK TM 60 hydrophobic protein analysis column to check the purity

[0068] Take the breakthrough peak C1 collected by the CM column and pass through Protein-PAK TM 60 Hydrophobic protein analysis column, the sample volume is 1μL, the mobile phase is water, and the electropherogram results show a single peak.

[0069] 2. SDS-PAGE electrophoresis to detect the purity of β-glucanase

[0070] Based on the existing SDS-PAGE technology, ultrafiltration membrane interception (No. 1 sample), ammonium sulfate precipitation 30%-65% (No. 2 sample), CM ion exchange peak C1 (No. 3, 4 samples), Q ion exchange breakthrough The peak Q1 (sample No. 5) was detected by 12% SDS-PAGE gel electrophoresis. The red box shows the β-glucanase produced by Bacillus marinum N6-2. See figure 2 .

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Abstract

The invention relates to a beta-glucanase generated by Bacillus marinus and a preparation method thereof, belonging to the fields of marine microbes and enzyme preparations. The beta-glucanase is generated by Bacillus marinus N6-2, and the molecular weight is 24kDa. The beta-glucanase has the advantages of higher enzyme activity, favorable heat stability and favorable pH stability, and has wide development potential and application prospects in the aspects of wine industry, industrial wastewater treatment, functional food development and feed application.

Description

technical field [0001] The invention belongs to the field of marine microorganisms and enzyme preparations, and in particular relates to a beta-glucanase produced by marine bacillus and a preparation method thereof. Background technique [0002] β-glucanase is a kind of hydrolase that can degrade β-glucan in grain and can be applied in beer production. The main component of the barley endosperm cell wall is β-glucan, which can degrade the β-1,3 and β-1,4 glycoside chains in the β-glucan molecule, making it degrade into small molecules and losing hydrophilicity , reduce the viscosity, thereby loosening the barley endosperm cell wall, increasing the utilization rate of barley, reducing the viscosity of wort, greatly shortening the filtration time of wort and beer, increasing the beer output and improving the quality of beer. There is a large amount of β-glucan in the feed of barley, wheat, rye, oats and other grains for livestock and poultry. Since there is no enzyme to diges...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12R1/07
CPCC12N9/244C12Y302/01006
Inventor 郑兰红康道乐杨康利梁方方朱美虹
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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