Bacillus subtilis strain and applications thereof

A technology of Bacillus subtilis and strains, applied in the direction of bacteria, biochemical equipment and methods, microorganisms, etc., can solve the problem of not being able to use glycerin, and achieve the effect of cheap raw materials, high output, and short fermentation cycle

Active Publication Date: 2014-06-18
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In view of the defect that existing Bacillus subtilis strains cannot use glycerol, especially crude glycerol, a by-product of biodiesel, to produce lipopeptide biosurfactin surfactin, therefore, the present invention separates and purifies a strain that metabolizes lipopeptide from

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  • Bacillus subtilis strain and applications thereof
  • Bacillus subtilis strain and applications thereof
  • Bacillus subtilis strain and applications thereof

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0034] Example 1

[0035] This example illustrates the obtaining of Bacillus subtilis G-34 ( Bacillus subtilis G-34) method.

[0036] It includes the following steps:

[0037] 1) Obtain biological surface active strains in petroleum-contaminated soil by enriching the medium.

[0038] Take samples of petroleum-contaminated soil from Shengli Oilfield, and inoculate the processed soil samples into enrichment medium to obtain enrichment culture solution. Taking advantage of the hemolytic properties of the active agent of the biological surface, the enriched culture solution is inoculated on the blood plate by streaking and coating separation method, and cultured at 37°C for 24-48h to obtain a single colony with a large hemolytic circle and a transparent.

[0039] The enrichment medium formula is: glucose 20 g / L, ammonium sulfate 1 g / L, sodium nitrate 2 g / L, magnesium sulfate 0.3 g / L, potassium dihydrogen phosphate 1 g / L, and disodium hydrogen phosphate 4 g / L, yeast powder 2 g / L.

[0040...

Example Embodiment

[0050] Example 2

[0051] This example illustrates the treatment of Bacillus subtilis strains Bacillus subtilis The result of preliminary identification of the type of surfactant produced by G-34.

[0052] To Bacillus subtilis Strain G-34 is a strain, and glycerol is used as a carbon source for shake flask fermentation. Bacillus subtilis The biosurfactant produced by G-34 was purified and analyzed by FT-IR.

[0053] The analysis results are attached figure 1 . In the IR spectrum, 3303cm-1 is the NH stretching vibration caused by the hydrogen bonds between molecular chains, and 1656cm-1 and 1550cm-1 are the thalidomide bands I and II. The hydrophilic group of the above-mentioned characteristic absorption surface surfactant molecule is a peptide chain. The two absorptions of the spectrum 2872-2960cm-1 and 1243-1402cm-1 are the CH stretching vibration table of the fatty acid family. The hydrophobic part of the surfactant molecule is a fat half molecule, indicating that the surfac...

Example Embodiment

[0054] Example 3

[0055] This example illustrates Bacillus subtilis Bacillus subtilis G-34 uses pure glycerol to ferment to produce lipopeptide biosurfactants.

[0056] (1) Preparation of seed culture medium: yeast extract 5 g / L, peptone 10 g / L, and NaCl 10 g / L. Sterilize at 121 ℃ for 20 min, and cool for later use.

[0057] (2) Preparation of fermentation medium: glycerol 20 g / L, NH 4 NO 3 2 g / L, KH 2 PO 4 3 g / L, Na 2 HPO 4 10 g / L, MgSO 4 ·7H 2 O 0.2 g / L, FeSO 4 ·7H 2 O 0.02 g / L.

[0058] The medium was adjusted to pH 7.0 with phosphoric acid (1mol / L), sterilized at 121°C for 20 minutes, and cooled for later use.

[0059] (3) Pick a ring Bacillus subtilis The G-34 strain was placed in the medium obtained in step (1); placed at 37° C. and cultured at 200 rpm for 12 hours to obtain seed liquid.

[0060] (4) Take 1 ml of the Bacillus subtilis G-34 seed solution obtained in step (3) and inoculate 50 ml of the fermentation medium obtained in step (2). Incubate at 37°C and 200 rpm.

...

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Abstract

The invention discloses bacillus subtilis. The bacillus subtilis which is classified and named as G-34(Bacillus subtilis G-34) is collected in China Center for Type Culture Collection on January 6, 2014, and the collection register number is CCTCC NO:M2014003. A bacillus subtilis strain is obtained via mutation breeding on Bacillus subtilis strain separated from polluted soil of Shengli Oil Field; a culture medium with glycerin as the only carbon source can be rapidly utilized for preparing a lipopeptide biosurfactant surfactin at a high yield. With crude glycerin as a substrate, the pure surfactin can be obtained through the steps of bacillus subtilis fermenting-transforming, centrifugal degerming, acid precipitating, freeze-drying, and methanol extracting.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to Bacillus subtilis G-34 ( Bacillus subtilis G-34), especially relates to the method of using the bacterial strain to transform glycerol and ferment to produce lipopeptide biosurfactant. Background technique [0002] Lipopeptides are a class of compounds composed of hydrophilic peptide chains and lipophilic aliphatic chains. It contains a peptide chain composed of 7-10 amino acids and a β-hydroxy fatty acid chain or a β-amino fatty acid chain, wherein the hydroxyl or amine group on the fatty acid chain combines with the carboxyl group on the amino acid of the peptide chain to form a lactone bond or amide bond, The peptide chains are closed to form a cyclic lipopeptide. Because lipopeptide has a special chemical composition and amphiphilic molecular structure, it has broad application prospects in the fields of medicine, food, cosmetics and microbial oil recovery, and has become a rese...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/00C12R1/125
Inventor 李霜刘强黄和江凌
Owner NANJING UNIV OF TECH
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