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Functional expression of the Spodoptera exigua chitinase to examine the virtually screened inhibitor candidates

Published online by Cambridge University Press:  22 May 2019

L. Zhang ,
Z. Guan Open the ORCID record for Z. Guan [Opens in a new window] ,
Z. Pan ,
H. Ge ,
D. Zhou ,
J. Xu  and
W. Zhang
L. Zhang
Affiliation:
State Key Laboratory of Biocontrol and School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
Z. Guan
Affiliation:
State Key Laboratory of Biocontrol and School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
Z. Pan
Affiliation:
State Key Laboratory of Biocontrol and School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
H. Ge
Affiliation:
Medical College, Qingdao University, No. 38 Dengzhou Road, Qingdao 266021, China
D. Zhou
Affiliation:
Guangdong Provincial Key Laboratory of New Technology in Rice Breeding, Rice Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China College of Life Sciences, South China Agricultural University, Guangzhou 510642, China
J. Xu
Affiliation:
Research Center for Drug Discovery, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China
W. Zhang*
Affiliation:
State Key Laboratory of Biocontrol and School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
*
*Author for correspondence Phone: + 86 20 39332963 Fax: +86 20 39943515 E-mail: lsszwq@mail.sysu.edu.cn
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Abstract

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 SeChi gene (ADI24346) identified in our laboratory, we constructed the recombinant baculovirus P-Chi for the expression of recombinant SeChi (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 SeChi 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 SeChi 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.

Keywords

Spodoptera exiguachitinasebaculovirus expression systemhomology modelinginhibitor screening
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 109 , Issue 6 , December 2019 , pp. 741 - 751
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Copyright
Copyright © Cambridge University Press 2019 

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Footnotes

†

The first two authors contributed equally to this work

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