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Acyl coenzyme A thioesterase, application of gene thereof in degradation of zearalenone and preparation for degrading zearalenone

A technology of zearalenone and thioesterase, applied in application, genetic engineering, plant gene improvement, etc., can solve the problems of unclear degradation mechanism and related functional genes

Active Publication Date: 2019-06-21
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the degradation mechanism and related functional genes of this strain are still unclear.

Method used

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  • Acyl coenzyme A thioesterase, application of gene thereof in degradation of zearalenone and preparation for degrading zearalenone
  • Acyl coenzyme A thioesterase, application of gene thereof in degradation of zearalenone and preparation for degrading zearalenone
  • Acyl coenzyme A thioesterase, application of gene thereof in degradation of zearalenone and preparation for degrading zearalenone

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Embodiment 1

[0049] In the application of the recombinant expression vector containing the acyl-CoA thioesterase gene in this embodiment in the degradation of zearalenone, the acyl-CoA thioesterase gene is inserted into the recombinant expression vector, and the acyl-CoA thioesterase gene It has the nucleotide sequence shown in SEQ ID NO.1, SEQ ID NO.3 or SEQ ID NO.5.

[0050] Example 1 of the Application of Microbes Containing Acyl-CoA Thioesterase Genes in the Degradation of Zearalenone

[0051] In the application of the microorganism containing the acyl-CoA thioesterase gene in this embodiment to degrade zearalenone, the microorganism contains a recombinant expression vector, and the acyl-CoA thioesterase gene is inserted into the recombinant expression vector, and the acyl-CoA thioesterase gene is inserted into the recombinant expression vector. A thioesterase gene has a nucleotide sequence as shown in SEQ ID NO.1, SEQ ID NO.3 or SEQ ID NO.5. The microorganism may be a recombinant bac...

experiment example

[0059] 1. Research process

[0060] 1.1 Materials used in the experiment

[0061] 1.1.1 Strains: Bacillus amyloliquefaciens H6 is a strain that was screened out early and has zearalenone degradation properties (as shown in CN106047749A). Expression strain BL21 (DE3) and prokaryotic expression vector pet-30a are commercial strains and vectors.

[0062] 1.1.2 Main reagents: Zebralenone Standard (Pribola), methanol, nutrient broth, qiaquick-pcr kit (Takara), rneasy mini kit (Takara), dntpstakara, RNase H (Takara), DNA polymerase I (Takara), dissolving Buffer, IPTG and Trition X-100 (Biotechnology BioEngineering (Shanghai) Co., Ltd.), Plasmid Extraction Kit and DNA Extraction Kit (Tiangen Biochemical Te) Technology (Beijing) Co., Ltd., T4-DNA Ligase, TAQ -DNA polymerase (Bao Bioengineering (Dalian) Co., Ltd.).

[0063] 1.2 Experimental methods and results

[0064] 1.2.1 Sample preparation: Select a single colony and inoculate into 50mL nutrient solution (250 mL Erlenmeyer flas...

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Abstract

The invention relates to application of acyl coenzyme A thioesterase and gene thereof in degradation of zearalenone, and a preparation for degrading the zearalenone, and belongs to the technical fieldof biodegradation of mycotoxins. According to microbial transcriptome sequencing analysis, it discovers that the acyl- coenzyme A thioesterase gene of the YBGC / FADM family in a bacillus amyloliquefaciens H6 strain is a key gene for degrading the zearalenone, the acyl coenzyme A thioesterase gene is expressed in escherichia coli, the protein is found to be soluble, and recombinant protein is obtained after purification. A zearalenone ELISA detection kit detects that the recombinant protein has the biological activity of efficiently degrading ZEN. Therefore, the acyl coenzyme A thioesterase andthe coding gene thereof can be used for degrading the zearalenone, and the acyl coenzyme A thioesterase can be used for preparing an enzyme preparation or a complex enzyme preparation for degrading the zearalenone.

Description

technical field [0001] The invention relates to the application of acyl-CoA thioesterase and its gene in degrading zearalenone and a preparation for degrading zearalenone, belonging to the technical field of mycotoxin biodegradation. Background technique [0002] According to the statistics of the Food and Agriculture Organization of the United Nations, about 25% of the world's crops are polluted by mycotoxins every year, and about 2% of the crops lose their nutritional and economic value due to severe pollution, causing hundreds of billions of dollars in economic losses. Excessive mycotoxins have become an important factor that endangers the quality and safety of agricultural products, causing huge economic losses to the grain, edible oil and feed industries, and seriously threatening the health and safety of humans and livestock. Zearalenone (ZEN) is an estrogen-like mycotoxin produced by many Fusarium species. Before and after harvesting corn, wheat and other grains, the...

Claims

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Application Information

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IPC IPC(8): A23L5/20C12N9/16C12N15/55
Inventor 王彦彬孙向丽许来鹏康相涛田亚东闫峰宾李转见
Owner HENAN AGRICULTURAL UNIVERSITY
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