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Bacillus amyloliquefaciens and preparation method of exopolysaccharides thereof

A technology of amylolytic spores and exopolysaccharides, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., and can solve problems such as the lack of varieties of microbial exopolysaccharides and the production that cannot meet market demand.

Active Publication Date: 2017-03-22
甘肃尚农生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the variety of microbial exopolysaccharides that has been put into industrial production in my country is not rich, and the output cannot meet the market demand of our country.

Method used

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  • Bacillus amyloliquefaciens and preparation method of exopolysaccharides thereof
  • Bacillus amyloliquefaciens and preparation method of exopolysaccharides thereof
  • Bacillus amyloliquefaciens and preparation method of exopolysaccharides thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Separation and Identification of Bacteria

[0039] Put the rhizosphere soil samples of Lily of Lanzhou into a Erlenmeyer flask filled with 25ml enrichment medium, and shake the flask at 30°C and 160r / min for 48h. Then, after diluting the enriched culture solution 10 times with sterile physiological saline, take 0.2 ml to coat the separation medium plate. Inverted cultivation at 28°C for 48 hours, colonies with a gelatinous transparent appearance on the plate were obtained, and bacteria capable of producing exopolysaccharides were obtained. Both the enrichment medium and the isolation medium are PDA plate medium. The 16S rDNA gene of the strain was sequenced according to conventional methods, and the sequencing results were compared with the sequence of Bacillus in Genebank, which showed the highest similarity, and the experimental strain was determined to be Bacillus amyloliquefaciens. It was identified and confirmed as Bacillus amyloliquefaciens by the Chi...

Embodiment 2

[0040] Example 2: Medium components and fermentation conditions of exopolysaccharide

[0041] LB medium was used as the seed medium; the composition of the fermentation sugar production medium was (g / L): sucrose 50.0. Yeast extract 5.0, peptone 10.0, (NH4) 2 SO 4 3.0, NaCl 3.0, MgSO 4 0.05, CaCl 2 0.05,K 2 HPO 4 1.0.

[0042] Fermentation conditions: liquid volume is 30mL culture medium per 100mL Erlenmeyer flask, inoculum size is 7.5% (v / v), initial pH value is 7.0, 32C, 160pm culture 12h.

Embodiment 3

[0043] Example 3: Crude and refined exopolysaccharide

[0044] (1) Strain activation: the preserved strains were inoculated into beef extract peptone slant medium, cultured at a constant temperature of 32°C for 24 hours, and stored in a refrigerator at 4°C for later use.

[0045] (2) Seed culture medium preparation and inoculation culture: the seed liquid culture medium adopts LB culture medium, and the filling amount is filled with 30mL culture medium for every 100mL triangular flask. The beef extract peptone slant strain in step (1) was inoculated on the seed culture medium, and cultured for 12 hours at a culture temperature of 30° C. and a rotation speed of 200 rpm to obtain a seed culture solution.

[0046] (3) Preparation and fermentation culture of fermentation sugar production medium: the composition of the fermentation sugar production medium is as above, and the pH value is 6.0. The seed solution fermented in step (2) was inoculated on the fermentation medium with an...

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Abstract

The invention provides bacillus amyloliquefaciens EZ99. The bacillus amyloliquefaciens is gram positive bacteria, exists in plant rhizosphere soil, has no pathogenicity for people and plants, is collected from an orchard county, in the west of a Qilihe district in the Lanzhou city of the Gansu province, and has been identified, registered and preserved in the China common microorganism strain preservation center, and the preservation number is CGMCC No.13267. Component analysis is performed on bacterial colonies formed by the bacillus amyloliquefaciens on a PMA and MS culture medium, and it is found that the bacillus amyloliquefaciens has the characteristic of synthesizing and secreting exopolysaccharides. Optimal culture medium formulas and fermentation optimal conditions based on fermentation production of the exopolysaccharides of the strains are built, a fermentation solution is subjected to centrifuging, alcohol precipitation, dialysis and chromatographic separation, four purification polysaccharide components are obtained, and four-polysaccharide-component monosaccharide composition and monomer molecular weight are determined. The exopolysaccharides obtained through the strain and the method have the various applications of medicines, cosmetics, food additives, biological film raw materials and the like.

Description

technical field [0001] The invention relates to the field of microbial fermentation products, in particular to a preparation method of Bacillus amyloliquefaciens and its exopolysaccharide. Background technique [0002] Exopolysaccharide is a kind of natural macromolecular substance synthesized by microorganisms and secreted into the environment. It is composed of multiple monosaccharide molecules dehydrated and polymerized, connected by various forms of glycosidic bonds, and can form straight or branched long chains. structure. The role of microbial exopolysaccharide itself is to create a suitable living environment for microorganisms. It participates in the formation of the extracellular matrix of microorganisms, which is the medium for microbial cells to attach to the environmental interface (such as sand, rock or plant surface), forming a microbial film (biofilm), so that it can obtain a relatively stable living environment. In addition, exopolysaccharides are also a wa...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/04C12R1/07
CPCC12P19/04C12N1/205C12R2001/07
Inventor 王若愚卢翔陈高峰
Owner 甘肃尚农生物科技有限公司
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