Molecular Nutrition

Expression of Zearalenone Degrading Enzyme in Bacillus subilis and Its Effects on Reproductive Performance of Sows

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  • 1. College of Veterinary Medicine, Gansu Agriculture University, Lanzhou 730070, China;
    2. Henan Academy of Agricultural Sciences, Key Laboratory of Animal Immunology of the Ministry of Agriculture, Zhengzhou 450002, China;
    3. College of Life Science and Technology, Gansu Agriculture University, Lanzhou 730070, China

Received date: 2017-04-11

  Online published: 2017-10-31

Abstract

The purpose of this study was to establish an expressing system of zearalenone (ZEN) degrading enzyme gene (ZEN-jjm) in Bacillus subtilis, and to determine the biodegradation activity and effect of its expression product on reproductive performance of sows. The ZEN-jjm gene was cloned and the recombinant expression plasmid was constructed by ligation cloned ZEN-jjm and pHT01 vector digested with EcoRⅠand NotⅠ enzymes, and then transformed into Bacillus subilis. Expression of ZEN-jjm protein in Bacillus subilis was analyzed by dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). And the degrading activity of ZEN-jjm protein was determined by high performance liquid chromatography (HPLC). The effects of ZEN degrading enzyme on reproductive performance of sows were evaluated. The results of double enzyme digestion and DNA sequencing demonstrated that ZEN-jjm was inserted into pHT01. The SDS-PAGE showed that one Bacillus subilis strain with high-level expression was obtained, and the size of the expressed protein was about 29 ku. The HPLC result illustrated that the expressed ZEN-jjm protein could effectively degrade ZEN. The evaluation results of reproductive performance of sows indicated that the expressed ZEN-jjm protein could significantly relieve the toxicity for sows caused by ZEN (P<0.05). In conclusion:1) the ZEN-jjm expression vector is successfully constructed and the ZEN-jjm protein is expressed in Bacillus subtilis in this study. 2) The expressed ZEN-jjm protein has the biological activity of degrading ZEN. 3) ZEN-jjm protein can significantly reduce the harm of ZEN on reproductive performance of sows when it is added in the feed.

Cite this article

WANG Xiangsheng, SUN Yaning, RUAN Chongmei, ZHANG Quanwei, ZHANG Yong, HU Xiaofei . Expression of Zearalenone Degrading Enzyme in Bacillus subilis and Its Effects on Reproductive Performance of Sows[J]. Chinese Journal of Animal Nutrition, 2017 , 29(11) : 4019 -4025 . DOI: 10.3969/j.issn.1006-267x.2017.11.023

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