Lipase producing strain tolerant to organic solvent as well as genes and applications of lipase

An organic solvent-resistant and organic solvent-resistant technology is applied in the fields of catalytic splitting of chiral compounds and organic solvent-resistant lipase production bacteria, which can solve the problems of low organic solvent tolerance and low enzyme yield, and achieve strong tolerance, The effect of high specific activity and high yield

Inactive Publication Date: 2011-09-07
NANJING UNIV OF TECH
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Problems solved by technology

Although there have been articles disclosing the organic solvent-resistant lipase produced by some extremophiles, the organic solvent tolerance is low and the enzyme production is low

Method used

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  • Lipase producing strain tolerant to organic solvent as well as genes and applications of lipase
  • Lipase producing strain tolerant to organic solvent as well as genes and applications of lipase
  • Lipase producing strain tolerant to organic solvent as well as genes and applications of lipase

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Embodiment Construction

[0023] 1. Screening procedure for natural strains of lipase resistant to organic solvents.

[0024] The following method was used for the initial screening: Vegetable oil was used as the only carbon source, and organic solvents such as cyclohexane, toluene, and DMSO at different concentrations were used as the screening pressure to obtain extremophiles resistant to organic solvents from oily soil samples. Olive oil rhodamine B plate medium was used, and the specific formula was: yeast extract 1 g / L, corn steep liquor 5 mL / L, K 2 HPO 4 1 g / L, MgSO 4 ·7H 2 O 0.5 g / L, olive oil 60 mL / L, Rhodamin B 0.024 g / L. The selected organic solvent-resistant microorganisms were inoculated on the olive oil rhodamine B plate, and the strains with high lipase production were initially screened according to the ratio of the colony to the size of the orange fluorescent circle. Six extremophile strains resistant to organic solvents with high lipase production were screened by this method.

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Abstract

The invention relates to a lipase producing strain tolerant to an organic solvent as well as genes of the lipase producing strain tolerant to the organic solvent and an application of the lipase producing strain tolerant to the organic solvent in catalyzing and splitting chiral compounds in an organic phase, belonging to the technical field of microbiology engineering and enzyme engineering. The classification name of the lipase producing strain is PseudomonasstutzeriLC2-8, the preservation registration number thereof is: CCTCC NO: M2020279. The lipase producing strain is a gram negative strain which can tolerate multiple organic solvents with a certain concentration. The lipase producing strain in the invention is split and cloned to the lipase coded genes tolerant to the organic solvent, which is produced by the lipase producing strain. The lipase tolerant to the organic solvent has the properties such as high yield, high specific activity, good tolerance to solvents, wide acting pH range and the like. The lipase also enjoys industrial application value such as catalyzing and splitting the chiral compounds in the organic phase and the like.

Description

technical field [0001] The invention belongs to the technical field of microbial engineering and enzyme engineering, and in particular relates to an organic solvent-resistant lipase-producing bacterium, its organic solvent-resistant lipase gene, and the organic solvent-resistant lipase's ability to catalyze and split chiral compounds in an organic phase application. Background technique [0002] Lipase (lipase, EC 3.1.1.3, triglyceride hydrolase) is a class of enzymes that can catalyze the hydrolysis of long-chain fatty acid glycerides into glycerol and long-chain fatty acids. It is an important class of hydrolytic enzymes, which are widely used in many industrial fields such as medicine, detergent, food and leather. [0003] Lipase can catalyze the hydrolysis of ester bonds in an aqueous environment, and can catalyze transesterification, esterification, alcoholysis, acidolysis and other reactions in an organic solvent environment. The substrates of lipase are often water-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/20C12N15/55C12P7/62C11D3/386C11D7/42C12R1/38
Inventor 何冰芳曹艳姚长进吴斌李文豪
Owner NANJING UNIV OF TECH
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