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Strain for generating phospholipase D with high and stable yield by utilizing physical and chemical mutation

A chemical mutagenesis, phospholipase technology, applied in biochemical equipment and methods, electric/wave energy treatment enzymes, microorganism-based methods, etc., can solve the problems of low enzyme activity and high price of PLD, and achieve simple and efficient operation. high effect

Inactive Publication Date: 2010-11-03
DALIAN UNIV OF TECH
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Problems solved by technology

[0007] The purpose of the present invention is to use physical and chemical mutagenesis to produce a stable and high-yield PLD strain, solve the problems of low PLD enzyme activity and high price in the prior art, and lay the foundation for further large-scale production and application of PLD

Method used

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Embodiment Construction

[0027] The fermented seed medium used in the present invention consists of: glucose 10.0g / L, yeast extract powder 20.0g / L, peptone 5.0g / L, K 2 HPO 4 2.0g / L, MgSO 4 ·7H 2 O 0.5g / L, pH7.0; culture conditions: 28°C constant temperature shaking culture for 24h.

[0028] The composition of the fermentation medium used is: peptone 20.0g / L, soluble starch 25.0g / L, MgSO4 ·7H 2 O0.5g / L, CaCO 3 1.0g / L, Tween 80 20.0g / L; the culture conditions are: the fermentation period is 7d, the fermentation temperature is 28°C, the initial pH value of the fermentation broth is 6.0, the inoculum size is 3%, and the liquid volume of the fermentation medium is 10mL / 100mL Erlenmeyer flask, the shaker speed is 200rpm.

[0029] The method for measuring the PLD enzyme activity is as follows: take 0.5 g of lecithin, add 1 mL of ether to make a 500 mg / mL solution, add 10 mL of water, shake in an ice bath until an emulsion is formed. Take 100 μL into a test tube, add 100 μL of 0.1mol / L sodium citrate...

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Abstract

The invention relates to a strain for generating phospholipase D with high and stable yield by utilizing physical and chemical mutation, belonging to the technical field of microbial mutation. The strain is characterized in that the phospholipase D is prepared by fermenting streptomyces chromofuscus of high yield phospholipase D, which is obtained by taking the streptomyces chromofuscus as an original strain and sequentially carrying out composite mutation of ultraviolet rays and lithium chloride and composite mutation of atmosphere cold plasmas and the lithium chloride on a monospore bacterial suspension thereof. The invention has the advantages that the streptomyces chromofuscus of the high yield phospholipase D is obtained through mutation screening, thereby providing a high yield strain with higher application value to industrial production of the phospholipase D, laying a good foundation for fermentation culture and application of the phospholipase D, and also providing a good and feasible method for microbial mutation breeding. The invention adopts a mutation and screening method, has the advantages of simple operation, high efficiency and the like, and has a practical value on screening the strain with an industrial application value.

Description

technical field [0001] The invention belongs to the technical field of microbial mutagenesis, and relates to the use of physical and chemical mutagenesis to produce a stable and high-yield phospholipase D bacterial strain. Background technique [0002] Natural phospholipids are good natural surfactants, but their application range is very limited due to the structure and composition. In order to improve the performance of phospholipids, people research and develop new phospholipid products through phospholipid modification technology. Modified phospholipids with special physiological effects have high commercial value and broad market prospects, which also makes the modification of phospholipids a hot research topic at home and abroad. People try to modify phospholipids by various methods. [0003] Due to the similarity of phospholipid structure, it is difficult to prepare high-purity single phospholipids by general physical and chemical modification methods. For example, t...

Claims

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Application Information

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IPC IPC(8): C12N15/01C12N13/00C12R1/465
Inventor 杨天奎赵紫薇牟英
Owner DALIAN UNIV OF TECH
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