Sphingomonas sp. TP-5 and method and application of same for producing welan gum

A technology of sphingomonas and welan gum, which is applied in the fields of biotechnology and biomaterials, can solve the problems of different fermentation production processes and post-extraction methods, and achieve high gel strength, good stability, and ideal growth Thickness effect

Inactive Publication Date: 2010-01-06
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, Kelco has become the only producer and supplier of Welan Gum in the world. In China, Nanjing University of Technology, Nanchang University and Hebei Xinhe Biochemical Co., Ltd. have been engaged in research in this area, but the strains used, the fermentation production process and the The post-extraction methods are all different, and there is no relevant report on the production of Welan gum

Method used

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  • Sphingomonas sp. TP-5 and method and application of same for producing welan gum
  • Sphingomonas sp. TP-5 and method and application of same for producing welan gum
  • Sphingomonas sp. TP-5 and method and application of same for producing welan gum

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Experimental program
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Effect test

Embodiment 1

[0035] Screening and breeding of the sphingomonas TP-5 (Sphingomonas sp. TP-5) strain provided by the invention.

[0036] Collect wastewater samples from a molasses factory in Jinnan District, Tianjin, take 10mL in 90mL glucose inorganic salt medium (medium composition: KH 2 PO 4 3.48, Na 2 HPO 4 12H 2 O 1.5, NH 4 8O 4 2. MgSO 4 ·7H 2 O 0.70, yeast extract 0.05; glucose 5%; distilled water, 1000ml; pH7.2; 105°C sterilization for 30min), placed in a 200r / min low-temperature shaker at 30°C for enrichment culture for 5 days, the diameter of the streaked screen was larger and Obvious viscous colony; further select the colony and transfer it to fresh 100mL glucose inorganic salt medium for subculture and domestication, and the culture conditions are the same as above; after 5 to 7 cycles of enrichment, TP-5 is finally obtained; draw repeatedly on the LB solid plate Line, pick a single colony and save it on a slant; take a ring of TP-5 colony and put it into a test tube c...

Embodiment 2

[0038] The morphological characteristics and physiological and biochemical characteristics of the Sphingomonas TP-5 (Sphingomonas sp.TP-5) bacterial strain provided by the present invention.

[0039] According to the experimental method of "Bergey's Manual of Systematic Bacteriology" (Vol. VIII), the Gram staining, cell size and shape, presence or absence of flagella and spores, growth temperature, growth pH range, and NaCl tolerance were detected. Catalase, oxidase, M.R. experiment, V-P experiment, indole production, nitrate reduction, starch hydrolysis, ammonia production experiment, urease, hydrogen sulfide production, gelatin hydrolysis, glucose fermentation, litmus milk test, decomposition of escin experiment.

[0040] Sphingomonas sp.TP-5 (Sphingomonas sp.TP-5) can grow light yellow, round colonies when cultured on LB plate at 30°C for two days, the colony size is 1-2 mm in diameter, the edges are neat, and the surface is smooth ( See figure 1 ). Colonies will grow lar...

Embodiment 3

[0042] PCR amplification and sequence determination of the 16S rRNA gene of the sphingomonas TP-5 (Sphingomonas sp. TP-5) strain provided by the present invention.

[0043] The 16S rRNA gene sequence characteristics of the sphingomonas TP-5 (Sphingomonas sp.TP-5) bacterial strain provided by the present invention: the TP-5 bacterial strain is inoculated in LB medium, 30 ℃ shaker culture (180rpm) 24 hours , collected by centrifugation, resuspended, added lysozyme and SDS to break the wall, extracted genomic DNA by phenol-chloroform method, and used upstream primer (5'-GAGAGTTTGATCCTGGCTCAG-3') and downstream primer (5'-AAGGAGGTGATCCAGCCGCA-3' ), use this pair of primers to amplify its 16S rDNA gene by PCR, and send the amplified primers to Dalian Bao Biological Company for sequencing. The PCR conditions are: 94°C, 5min; 94°C, 45s, 56.2°C, 45s, 72°C, 90s, 30 cycles; 72°C, 9min, 4°C storage. The length of the 16S rDNA gene sequence is 1610p, and the sequence similarity with the ...

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Abstract

The invention relates to Sphingomonas sp. TP-5 and a method and an application of the same for producing welan gum. A strain of the invention is prepared by separating and acclimating sugar-contained wastewater in a molasses plant, has a systematic name of Sphingomonas sp. and a preservation number of CGMCCNo.3097, not only can grow in nutrient media of beef broth, LB and nutrient agar, but also can grow in a sugar-contained inorganic-salt culture medium. The TP-5 strain can be fermented in the sterile culture medium containing sugar, inorganic salt and water at a temperature condition of 28-37 DEG C; ymotic fluid is extracted to obtain the welan gum achieving a temperature-resistant limit value of 150 DEG C; and potassium chloride in different concentrations is added so that a welan gum solution remarkably improves the viscosity, enhances the temperature stability, does not vary the viscosity with temperature and presents stronger temperature tolerance, thereby being applied to seawater drilling mud and a high salinity reservoir viscous water flooding system.

Description

【Technical field】 [0001] The invention belongs to the technical field of biotechnology and biomaterials, in particular, it relates to a strain of Sphingomonas TP-5 (Sphingomonas sp. application. 【Background technique】 [0002] Many microorganisms can produce a certain amount of various polysaccharide polymers under different external conditions during their growth and metabolism. This kind of polysaccharide polymer is safe and non-toxic, and has unique physical and chemical properties. Compared with plant and animal polysaccharide polymers, the production of microbial polysaccharide polymers is less affected by factors such as geographical environment, climate, and natural disasters. The yield and quality All are stable. Welan gum is easily soluble in water, and its aqueous solution has unique rheological properties and good viscosity increasing, acid and alkali resistance. The temperature tolerance of its aqueous solution can be as high as 150°C, which is higher than the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/04C09K8/035C09K8/588C12R1/01
Inventor 马挺李国强李丹萍梁凤来刘如林
Owner NANKAI UNIV
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