Strain and method for producing Welan gum

A technology of welan gum and bacterial strains, which is applied in the field of industrial microorganisms, can solve the problems of different production strains and fermentation processes, and achieve the effects of inhibiting degradation, simple process, and efficient production

Active Publication Date: 2015-08-19
CHINA UNIV OF PETROLEUM (EAST CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, welan gum is mainly produced through microbial fermentation. The American kelco company has become the largest producer and supplier of welan gum in the world. Domestic research on welan gum is still in its infancy. Nanjing University of Technology, Nanchang University, Jiangnan The University and Hebei Xinhe Chemical Co., Ltd. are engaged in research in this area, but the production strains and fermentation processes are different, and there are no other related reports

Method used

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  • Strain and method for producing Welan gum
  • Strain and method for producing Welan gum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Sphingomonas provided by the invention Sphingomonas Morphological and physiological and biochemical identification of sp.WG strain

[0031] Sphingomonas WG provided by the invention ( Sphingomonas sp.WG) was isolated from the sea mud of Jiaozhou Bay, Qingdao, and the pure culture was obtained after serial dilution and plate plating. The strain morphology and physiological and biochemical reactions were identified by Gram staining, morphological observation, and Bojian Gram-negative bacteria identification system.

[0032] Sphingomonas WG ( Sphingomonas sp.WG) cultured on LB plates at 30°C for 24 hours to grow a single colony, the single colony was yellow, round, convex in the center, viscous, opaque, with neat edges and smooth surface (see attached figure 1 ); the strain is Gram-staining negative, the thallus is short rod-shaped, has flagella, and has no spores; its growth temperature is 20-40°C, growth pH is 4-9, and NaCl tolerance is 3%; oxidase, Arg...

Embodiment 2

[0033] Example 2 Sphingomonas WG provided by the invention ( Sphingomonas sp.WG) cloning and sequencing of the 16S rRNA gene

[0034] Sphingomonas WG provided by the invention ( Sphingomonassp.WG) 16S rRNA gene sequence characteristics: the WG strain was inoculated in LB medium, cultured at 30°C and 180 rpm for 16-18 h, and its genomic DNA was extracted using the Bacterial Genomic DNA Extraction Kit (Tiangen Biochemical Technology Co., Ltd.). The 16S rRNA gene fragment was obtained by PCR amplification using the upstream primer 5'-AGAGTTTGATCMTGGCTCAG-3' and the downstream primer 5'-TACGGYTACCTTGTTACGACTT-3'. The PCR conditions were as follows: 94°C for 5 min; 30 cycles of 94°C for 50 s, 56°C for 45 s, and 72°C for 90 s; 72°C for 10 min. After the 16S rRNA gene fragment obtained by PCR amplification was purified, it was connected to the pMD18-T vector with the quick connection kit, and the connection product was transformed into E. coli DH5α was screened on LB plates co...

Embodiment 3

[0035] Example 3 Preparation of seed solution

[0036] Solid medium: yeast extract 10 g / L, peptone 20 g / L, glucose 20 g / L, agar 20 g / L, pH7.5.

[0037] Liquid medium: yeast extract 10 g / L, peptone 20 g / L, glucose 20 g / L, pH 7.5.

[0038] The strain of Sphingomonas WG was streaked on YPD solid medium and cultured at a constant temperature of 30°C for 4 days to be activated. When yellow colonies grew, it was stored at 2-4°C for a short period of time until use. The activated single colony was transferred into a 250 ml Erlenmeyer flask filled with YPD liquid medium, and the volume of the Erlenmeyer flask was 50 ml. It was incubated at 30°C and the shaker speed was 230 rpm for 18 h to obtain the first-grade seed liquid. The primary seed liquid was inoculated into a 500 ml Erlenmeyer flask containing 100 ml liquid YPD medium at a 1.5% inoculation amount, and incubated at 30°C for 15 h at a shaker speed of 230 rpm to obtain a secondary seed liquid.

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Abstract

The invention relates to a Welan gum producing marine strain, which belongs to Sphingomonas sp.WG, and has a preservation number of CCTCC No: M2013161. According to the present invention, the strain can be cultured in a liquid culture medium so as to obtain the Welan gum, and the production efficiency is high; the requirements of the strain of the present invention on the carbon source and the nitrogen source are low; the fermentation process comprises: (1) seed liquid culture and (2) fermentation production of Welan gum; the extraction and purification steps comprise: (1) fermentation broth pretreatment, (2) crude product precipitation, (3) impurity removing, and (4) drying so as to obtain the Welan gum product; with application of the marine strain to produce the Welan gum in the fermentation manner, the yield can be up to 33 g / L, and the extraction and purification yield can be up to 95%; and with the strain and the method of the present invention, the low-cost, high-efficiency and high-quality Welan gum production can be achieved.

Description

technical field [0001] The present invention relates to a marine bacterial strain Sphingomonas ( Sphingomonas sp. WG) and a process for high-yield fermentation and extraction and purification of welan gum using the strain, belonging to the technical field of industrial microorganisms. Background technique [0002] Welan gum (Welan gum) is a heteropolysaccharide synthesized by microorganisms. It is a high molecular polymer. Its skeleton structure consists of D-glucose, D-glucuronic acid, D-glucose and L-rhamnose. unit composition. The side chain is composed of single-chain L-mannose or single-chain L-rhamnose. [0003] Welan gum has good thermal stability, suspending, emulsifying and thickening properties, and can be used as a thickener, suspending agent, emulsifying agent, stabilizer, lubricant, film-forming agent and adhesive in industrial and agricultural applications. It has broad application prospects in various aspects, especially in industries such as food, concrete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/04C08B37/00C12R1/01
CPCC08B37/00C12N1/20C12P19/04C12N1/205C12R2001/01
Inventor 朱虎李慧冯志梅周万龙孙亚杰吕建仁
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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