Pesticide degradation enzyme and preparation method thereof

A pesticide-degrading enzyme and a technology for degrading enzymes are applied in the field of pesticide-degrading enzyme agent and its preparation, which can solve problems such as threats to food safety, deterioration of agricultural production environment, etc., and achieve the effects of ensuring food safety, no toxic side effects, and high enzyme activity.

Active Publication Date: 2013-03-27
辽宁中科生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Pesticide residues lead to the deterioration of the agricultural production en

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Inoculate the genetically engineered strains preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee, preservation number: CGMCC No.1529, in LB liquid medium containing ampicillin, culture temperature 37 ° C, dissolved oxygen 20%, culture time 10 Hours. LB liquid medium is calculated by weight percentage, tryptone 1%, yeast powder 0.5%, sodium chloride 1%, the balance is distilled water, sterilized at 121 ℃ for 20 minutes; LB liquid medium containing ampicillin is LB Add 50 μg of ampicillin to the liquid medium.

[0018] The genetically engineered strain is preserved in the General Microbiology Center of China Committee for the Collection of Microbial Strains, with a preservation number of CGMCC No. 1529. Also refer to the relevant records in the application number 200510086957.3.

[0019] Then the culture solution is inoculated in the fermentation medium containing ampicillin by the inoculum amount of 1% by volume,...

Embodiment 2

[0023] The genetically engineered bacteria were inoculated in LB liquid medium containing ampicillin, the culture temperature was 37°C, the dissolved oxygen was 20%, and the culture time was 10 hours. LB liquid medium is calculated by weight percentage, tryptone 1%, yeast powder 0.5%, sodium chloride 1%, the balance is distilled water, sterilized at 121 ℃ for 20 minutes; LB liquid medium containing ampicillin is LB Add 50% ampicillin to the liquid medium.

[0024] Then the culture solution is inoculated on the fermentation medium containing ampicillin by the inoculation amount of 1% by volume, at 37 DEG C, the dissolved oxygen amount is 25%, and after cultivating for 3 hours, adding final concentration in the fermentation culture medium is 25mmol / L After lactose, the culture temperature was lowered to 25°C, and after 22 hours of induction expression, the fermentation broth was centrifuged at 5000rpm for 10 minutes to collect the bacterial cells, and 0.1mol / L pH 7.0 phosphate b...

Embodiment 3

[0028] The genetically engineered bacteria were inoculated in LB liquid culture medium containing ampicillin, the culture temperature was 37°C, the dissolved oxygen was 20%, and the culture time was 10 hours. LB liquid medium is calculated by weight percentage, tryptone 1%, yeast powder 0.5%, sodium chloride 1%, the balance is distilled water, sterilized at 121 ℃ for 20 minutes; LB liquid medium containing ampicillin is LB Add 50% ampicillin to the liquid medium.

[0029] Then the culture solution is inoculated on the fermentation medium containing ampicillin by the inoculation amount of 2% by volume, at 37 DEG C, the dissolved oxygen amount is 30%, and after cultivating for 3 hours, adding final concentration in the fermentation culture medium is 25mmol / L After lactose, the culture temperature was lowered to 22°C, and after 20 hours of induction expression, the fermentation broth was centrifuged at 5000rpm for 10 minutes to collect the bacterial cells, and 0.1mol / L pH 7.0 pho...

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Abstract

The invention relates to the technical field of an enzyme preparation technology, and particularly relates to a pesticide degradation enzyme and a preparation method thereof. The pesticide degradation enzyme is the supernate obtained by the broken thalli after fermentation culture of a genetic engineering strain which is collected in the China General Microbiological Culture Collection Center and the collection number of which is CGMCC No.1529. Through the adoption of the invention, the prepared pesticide degradation enzyme can degrade the organophosphorus pesticide residues and remove pesticide residues on the surfaces of fruits and vegetables to guarantee food safety.

Description

technical field [0001] The invention relates to the technical field of enzyme preparation technology, in particular to a pesticide degrading enzyme and a preparation method thereof. Background technique [0002] Pesticide residues lead to the deterioration of the agricultural production environment and threaten food safety, which has always been a concern at home and abroad. The biodegradation of pesticide residues is currently a research hotspot at home and abroad. Using genetic engineering to construct pesticide detoxifying enzyme genetically engineered bacteria, fermenting and expressing pesticide degrading enzymes is an effective technical means to solve the problem of pesticide residues. It is safe, efficient and unique. Secondary pollution and other characteristics. [0003] The preparation process of pesticide-degrading enzymes is directly related to the level of enzyme activity. The process of preparing pesticide-degrading enzymes using genetically engineered bacter...

Claims

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Application Information

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IPC IPC(8): C12N9/02A62D3/02A62D101/04A62D101/26
Inventor 谢建飞石元亮卢宗云张惠文李鹏程邢云鹏李敏
Owner 辽宁中科生物工程股份有限公司
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