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High-density fermentation method and product storage method of organophosphorus hydrolase recombinant strains

A high-density fermentation and hydrolase technology, applied in the field of environmental microorganisms and agriculture, to achieve the effect of maintaining enzyme activity and prolonging shelf life

Inactive Publication Date: 2010-07-21
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

Although chemical reactions can also transform the structure of organic pollutants, these transformations are usually subtle and the products are often structurally and toxically similar to the parent compound

Method used

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  • High-density fermentation method and product storage method of organophosphorus hydrolase recombinant strains
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  • High-density fermentation method and product storage method of organophosphorus hydrolase recombinant strains

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Embodiment Construction

[0022] (1) Optimization of fermentation medium and fermentation conditions

[0023]The bacterial strain used is Eshcherichia coli BL21 (DE3) pETMb (publicly known, Guoping Fu, Zhongli Cui, Tingting Huang and Shunpeng Li, Expression, purification, and charac-terization of a novel methyl parathion hydrolase. Protein Expression and Purification, 2004, 36: 170-176). The expression rate of organophosphate hydrolase gene mpd (encoding MPH protein) in Escherichia coli is much higher than the rate of post-translational processing. Due to the hydrophobicity caused by the signal peptide structure, most of the expression products are precipitated before being processed. The expression strain is not suitable for the preparation of organophosphate hydrolase engineering strains. Eshcherichia coli BL21(DE3)pETMb is obtained by removing the mpd signal peptide coding region through genetic engineering and constructing the expression plasmid pET29a(+) to obtain the expression plasmid pETMb, tr...

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Abstract

The invention relates to a high-density fermentation method and a product storage method of organophosphorus hydrolase recombinant strains, which belong to the technical field of environmental biotechnology. The invention optimizes the high-density culture conditions of organophosphorus hydrolase MPH recombinant strains Eshcherichia coliBL2l (DE3) pETMb, so that the optical density of fermentation fluid OD after 12 hours of fermentation reaches 29, and the enzyme activity reaches 18.5IU / ml. In addition, appropriate protective solution is added in the MPH production, so as to prolong the storage life of the enzyme, and the enzyme can also be mixed with a bio-surfactant, and simplify the use procedure of degrading enzyme. The invention can serve as an optimized formula and be applied to the production of fruits and vegetables detergent with enzyme, so that the invention plays a greater role in the pesticide degradation on the surface of fruits and vegetables. Through the storage scheme of the invention, the storage life of the enzyme can be prolonged to above 90 days under the storage conditions of liquid.

Description

technical field [0001] The invention relates to a high-density fermentation method for producing bacterial strains of organophosphorus hydrolase MPH and liquid preservation conditions of organophosphorus hydrolase MPH, belonging to the fields of applied environmental microorganisms and agriculture. Background technique [0002] Pesticides are the material guarantee of agricultural production. Their use greatly reduces the loss of crops and the occurrence of insect-borne diseases, and improves the economic benefits of agricultural production. With the continuous growth of population, the demand for food will become more and more tense, and the use of pesticides will be inevitable in a long period of time. Since 1990, my country's pesticide production ranks second in the world. Since the 1980s, the sales of organophosphorus pesticides have always ranked first among all kinds of pesticides. Organophosphorus pesticides are irreversible inhibitors of acetylcholinesterase, which...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/16C12N9/96C12R1/19
Inventor 崔中利曹慧林恒顾海莎李顺鹏
Owner NANJING AGRICULTURAL UNIVERSITY
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