EXPERIMENTAL METHOD AND ANIMAL

Screening and Effect Evaluation of Daphnetin as an Inhibitor of Ruminal Microbial Urease

  • HE Yue ,
  • ZHAO Shengguo ,
  • ZHANG Xiaoyin ,
  • ZHENG Nan ,
  • WANG Jiaqi
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  • 1. State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
    2. Laboratory of Quality and Safety Risk Assessment for Dairy Products of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received date: 2021-01-21

  Online published: 2021-09-18

Abstract

This study was designed to obtain an urease inhibitor that belonged to plants, and then to explore the basic characteristics of its inhibition to ruminal microbial urease. Firstly, selected plant-sourced compound library was screened via molecular docking with ruminal microbial urease as target, combining ruminal microbial urease inhibition assay to gain the compound with the best inhibition. And then, The half inhibitory concentration (IC50), enzyme kinetic characteristics and acting sites of this compound to ruminal microbial urease were test. Finally, using anaerobic culture test of rumen microorganism in vitro to evaluate its influence to urea. The results showed as follows:1) daphnetin was the compound with the best inhibition under the same concentration. 2) The IC50 of daphnetin to ruminal microbial urease was (93.43±3.87) μmol/L. What's worse, daphnetin was mixed urease inhibitor to ruminal microbial urease. 3) With the interact of thiol reagents, the inhibitory capacity of daphnetin to ruminal microbial urease had decreased significantly (P<0.05), and molecular docking results indicated that daphnetin could interact with amino acid Ala-166, His-245, Lys-216, Ala-362 and Cys-318. 4) Daphnetin could decreased decomposition of urea of anaerobic culture system for rumen microorganisms in vitro markedly after addition of 2, 4 and 6 h (P<0.05). In conclusion, daphnetin inhibits ruminal microbial urease through interacting with some amino acids belong to active site of urease, and reduces urea decomposition.

Cite this article

HE Yue , ZHAO Shengguo , ZHANG Xiaoyin , ZHENG Nan , WANG Jiaqi . Screening and Effect Evaluation of Daphnetin as an Inhibitor of Ruminal Microbial Urease[J]. Chinese Journal of Animal Nutrition, 2021 , 33(9) : 5256 -5265 . DOI: 10.3969/j.issn.1006-267x.2021.09.045

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