Bacterial cellulose producing strain, screening method and preparation of bacterial cellulose

A bacterial cellulose and screening method technology, applied in the field of microorganisms, can solve problems such as difficulty in determining the limitations of adjacent species, and achieve the effects of meeting the needs of industrial purification of bacterial cellulose, fast growth, and wide utilization of carbon sources

Active Publication Date: 2021-06-15
EAST CHINA NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the current identification of BC-producing strains is mainly through the combination of 16S rDNA and physiological and biochemical methods. This method has the limitation that it is difficult to determine the adjacent species. Exploring a new method for accurately classifying BC-producing strains will be of great help to the subsequent strains. The transformation or research of the

Method used

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  • Bacterial cellulose producing strain, screening method and preparation of bacterial cellulose
  • Bacterial cellulose producing strain, screening method and preparation of bacterial cellulose
  • Bacterial cellulose producing strain, screening method and preparation of bacterial cellulose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1: Screening of cellulose-producing strains

[0079] Use sterile surgical tweezers to pick out the pulp of decayed tissues from natural rotten grapes, kiwi, pears, dates, oranges, blackberry, oranges, plums, apples, strawberries, and watermelons, and add them to a sterile medium containing a primary enrichment medium. In glass test tubes, cultured at 30°C, if a gel-like film grows on the surface of the culture medium and the film is not easily broken after shaking, it is positive. Clamp the film, wash it with sterile normal saline for 3 times, then shake it in a sterile glass test tube filled with sterile normal saline for 10 minutes, apply gradient dilution on HS plate culture medium, and incubate it upside down at 30°C for 3 days. Pick a single colony, insert it into a small test tube filled with HS liquid medium, and culture it statically at 30°C, and a gel-like film grows on the liquid surface. Clamp the film, wash it three times with sterile saline, stick ...

Embodiment 2

[0080] Embodiment 2: Morphological observation of cellulose-producing strains

[0081] Plate colony morphology: static culture at 30°C, cultured on HS plate medium for 3 days, the colony characteristics of 171129Z2-3 colonies of Oliformis 171129Z2-3 are: milky yellow, round, smooth surface, convex, neat edges, 1mm in diameter, easy to pick take (see figure 1 ). The colonies formed cellulose blocks on the surface of the culture medium with the extension of culture time.

[0082] Bacteria morphology: Observing the Bacillus omae 171129Z2-3 through an optical microscope, the bacterium is a Gram-negative bacterium. 0.46 μm (see figure 2 ), which is not motile.

Embodiment 3

[0083] Example 3: Physiological and biochemical identification of cellulose-producing strains

[0084] Pick a single colony cultured on HS plate medium at 30°C for 3 days, insert it into HS liquid medium, culture it with shaking at 30°C for 20 hours, and draw out the bacterial liquid for physiological and biochemical identification. The detailed physiological and biochemical characteristics are shown in Table 1. Bacillus 171129Z2-3 can utilize a wide range of carbon sources.

[0085] Table 1 Physiological and biochemical identification results of Bacillus omae 171129Z2-3

[0086]

[0087] Table 2 Physiological and biochemical comparison between 171129Z2-3 and Komagataeibacter species sequence: 1, 171129Z2-3; 2, K. sacchari; 3, K. entanii; 4, K. europaeus; 5, K. intermedius 6, K. obediens; 7, K. hansenii; 8, K. xylinus; 9, K. swingsii; 10, K. rhaeticus; :+, positive; -, negative; NR, not reported;

[0088]

[0089]

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Abstract

The invention discloses a bacterial cellulose producing strain and an identification method thereof. The strain is named as Komagataeibacter sp.171129Z2-3, the strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.17276. The strain is gram-negative and short-rod-shaped bacteria, has the characteristics of wide selection range of a carbon source and a nitrogen source, high growth speed, strong thallus activity, strong cellulose production capacity and the like, can produce a large amount of bacterial cellulose during static culture and shake culture, and can achieve the yields of 5.12 g / L and 2.571 g / L after 4 days of fermentation respectively. Through physiological and biochemical characteristics, 16SrDNA, dnaK, groEL and rpoB gene sequence analysis and genome average nucleotide index (ANI) analysis, the strain is determined to be foal bacillus. The strain has a good implementation prospect in the aspect of industrial production of the bacterial cellulose.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to the isolation and identification of a bacterial cellulose producing strain and the dynamic oscillation fermentation culture of bacterial cellulose. Background technique [0002] Bacterial Cellulose (BC) is fermented by microorganisms to form cellulose. It is a parallel straight-chain molecule formed by the polymerization of glucose through β-1,4-glucosidic bonds. It has no branch structure. The hydrogen bonds between molecules and molecules form a biological macromolecule with a three-dimensional network structure. It belongs to the primary microbial A kind of metabolite, which mainly plays the role of maintaining the shape of bacteria and protecting the bacteria from external damage. Compared with plant cellulose, BC does not contain lignin and semi-lignin, which makes it have a high degree of chemical purity, crystallinity (80-90%), water absorption capacity and d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12P19/04C08B15/00C12R1/01
CPCC12N1/20C12N1/02C12P19/04C08B15/00Y02E50/10
Inventor 易正芳高红亮陆婷芬黄婕蒋德明刘明耀
Owner EAST CHINA NORMAL UNIV
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