Functional expression of the Spodoptera exigua chitinase to examine the virtually screened inhibitor candidates
文献类型： 外文期刊
作者： Zhang, L. 1 ; Guan, Z. 1 ; Pan, Z. 1 ; Ge, H. 3 ; Zhou, D. 4 ; Xu, J. 6 ; Zhang, W. 1 ;
作者机构： 1.Sun Yat Sen Univ, State Key Lab Biocontrol, Guangzhou 510275, Peoples R China
2.Sun Yat Sen Univ, Sch Life Sci, Guangzhou 510275, Peoples R China
3.Qingdao Univ, Med Coll, 38 Dengzhou Rd, Qingdao 266021, Peoples R China
4.Guangdong Acad Agr Sci, Rice Res Inst, Guangdong Prov Key Lab New Technol Rice Breeding, Guangzhou 510640, Peoples R China
5.South China Agr Univ, Coll Life Sci, Guangzhou 510642, Peoples R China
6.Sun Yat Sen Univ, Res Ctr Drug Discovery, Sch Pharmaceut Sci, Guangzhou 510006, Peoples R China
关键词： Spodoptera exigua; chitinase; baculovirus expression system; homology modeling; inhibitor screening
期刊名称：BULLETIN OF ENTOMOLOGICAL RESEARCH （ 影响因子：1.75； 五年影响因子：2.007 ）
ISSN： 0007-4853
年卷期： 2019 年 109 卷 6 期
页码：
收录情况： SCI
摘要： Chitinase is responsible for insect chitin hydrolyzation, which is a key process in insect molting and pupation. However, little is known about the chitinase of Spodoptera exigua (SeChi). In this study, based on the Se Chi gene (ADI24346) identified in our laboratory, we constructed the recombinant baculovirus P-Chi for the expression of recombinant Se Chi (rSeChi) in Hi5 cells. The rSeChi was purified by chelate affinity chromatography, and the purified protein showed activity comparable with that of a commercial SgChi, suggesting that we harvested active Se Chi for the first time. The purified protein was subsequently tested for enzymatic properties and revealed to exhibit its highest activity at pH 8 and 40 C. Using homology modeling and molecular docking techniques, the three-dimensional model of Se Chi was constructed and screened for inhibitors. In two rounds of screening, twenty compounds were selected. With the purified rSeChi, we tested each of the twenty compounds for inhibitor activity against rSeChi, and seven compounds showed obvious activity. This study provided new information for the chitinase of beet armyworm and for chitinase inhibitor development.
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