Sphingomonas paucimobilis strain and application thereof

A technology of less motile sheaths and unicellular bacteria, applied in the field of bioengineering, can solve problems such as non-commercial production, and achieve the effects of convenient operation and simple nutrients

Inactive Publication Date: 2016-04-20
GUANGDONG ZHONGKE TIANYUAN NEW ENERGY SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, only Kelco company in the United States commercially produces welan gum, and there is no repo

Method used

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  • Sphingomonas paucimobilis strain and application thereof
  • Sphingomonas paucimobilis strain and application thereof
  • Sphingomonas paucimobilis strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Isolation and screening of Sphingomonas paucimobilis (Sphingomonas paucimobilis) Y5

[0032] Separation: Take the activated sludge sample from the alcohol wastewater plant, make a 10-fold gradient dilution with sterile water, then spread it on the LB agar plate, and culture it at 30°C.

[0033] Primary screening: Pick large, viscous, smooth and moist single colonies for secondary screening.

[0034] Re-screening: The single colonies picked by the primary screening were inoculated in liquid medium (glucose 30g / L, peptone 3g / L, yeast powder 1g / L, K 2 HPO 4 ·7H 2 O2g / L, MgSO 4 ·7H 2 (20.2g / L), 30 DEG C, 200r / min shaker culture, finally screen out the bacterial strain Sphingomonas paucimobilis (Sphingomonas paucimobilis) Y5 that fermented liquid viscosity and crude sugar output are high.

Embodiment 2

[0035] Example 2 Identification of Sphingomonas paucimobilis (Sphingomonas paucimobilis) Y5

[0036] 1) Colony morphology and microscopic examination, the results are as follows figure 2 Shown:

[0037] On the nutrient agar (NA) plate, the colony shape is round, yellow, moist, protruding growth, and the diameter of the colony is 1-5 mm; the microscopic examination shows Gram-negative bacteria.

[0038] 2) Oxidase and contact enzyme test:

[0039] The oxidase test was positive and the contact enzyme test was negative.

[0040] 3) 16srDNA identification:

[0041]Use an inoculation loop to pick 2-5 colonies and transfer them to TE, incubate at 37°C for 1.5h, add NaCl and CTAB solution, mix well, incubate at 65°C for 10min, add phenol-chloroform-isoamyl alcohol solution for extraction, and centrifuge for 10min , transfer the supernatant to a new EP tube, extract twice with chloroform-isoamyl alcohol, transfer to a new EP tube, add isopropanol to precipitate DNA, let stand at ...

Embodiment 3

[0046] Example 3 Detection and analysis of microbial polysaccharides produced by fermentation of Sphingomonas paucimobilis Y5.

[0047] The polysaccharide polymer produced by the fermentation of the strain is soluble in water, but insoluble in organic solvents such as alcohol and ketone.

[0048] The glucuronic acid content measured in the polysaccharide sample by the phenol-sulfuric acid method and the sulfuric acid-carbazole method is 12%; the polysaccharide is composed of D-glucose, D-glucose Aldehydic acid, D-mannose and L-rhamnose composition; Measure mannose in the analysis sample by gas chromatography: glucose: rhamnose=1:3.0:4.5; Determinate glycosidic bond in the polysaccharide by periodic acid oxidation method The types are mainly 1→4 keys and 1→3 keys.

[0049] The above experimental results show that the polysaccharide polymer belongs to welan gum.

[0050] Send the above-mentioned Sphingomonas paucimobilis to the China Type Culture Collection Center for preserva...

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Abstract

The invention discloses a Sphingomonas paucimobilis strain and application thereof and belongs to the technical field of bioengineering. The Sphingomonas paucimobilis strain is named Sphingomonas paucimobilis strain Y5, is collected in CCTCC (China Center for Type Culture Collection), which is located in Wuhan University, Wuhan, China, and has the collection number of CCTCC NO: M 2015523. Welan gum can be prepared from the train by fermenting under appropriate conditions through carbon sources, nitrogen sources, inorganic salts and water . The strain disclosed by the invention has relatively high sugar conversion ratio and welan gum yield and has a certain application value. The welan gum produced through fermenting the strain is high in quality and is resistant to acids and bases, a 1% aqueous solution viscosity of the welan gum reaches 3500mPa.S, and the viscosity is fundamentally unchanged at the temperature of 150 DEG C. During the fermentation of the strain, the sugar conversion ratio is relatively high and reaches 55% to 60%, and the yield is relatively high and reaches 25-32g/L (dry weight).

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a strain of Sphingomonas paucimobilis and its application in fermenting and producing welan gum. Background technique [0002] Welan gum (welangum No. S-130) is a microbial heteropolysaccharide synthesized by certain Alcaligenes sp. or Sphingomonas sp. One of the third-generation microbial exopolysaccharides developed after raw gum and gellan gum, its molecular weight is as high as several million. Its structural skeleton is composed of D-glucose, D-glucuronic acid, D-glucose and L-rhamnose repeating units, and the side chain is composed of a single L-mannose or L-rhamnose. The structure is as follows: figure 1 As shown; Welan gum aqueous solution has pseudoplastic fluid characteristics, and its low-concentration aqueous solution has high viscosity and good stability. Xanthan gum is 20-30°C higher. The aqueous solution of welan gum is stable to acid and alka...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/04C12R1/01
Inventor 梁锏文李智斌张伟强文兵周宏才
Owner GUANGDONG ZHONGKE TIANYUAN NEW ENERGY SCI & TECH
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