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Biochemical characterization a digestive trypsin in the midgut of large cabbage white butterfly, Pieris brassicae L. (Lepidoptera: Pieridae)

Published online by Cambridge University Press:  07 November 2017

A. Sharifloo ,
A. Zibaee ,
J. Jalali Sendi  and
K. Talebi Jahroumi
A. Sharifloo
Affiliation:
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht 416351314, Iran
A. Zibaee*
Affiliation:
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht 416351314, Iran
J. Jalali Sendi
Affiliation:
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht 416351314, Iran
K. Talebi Jahroumi
Affiliation:
Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
*
*Author for correspondence Phone:+98-013-33690485 Fax:+98-013-33690281 E-mail: arash.zibaee@guilan.ac.ir, arash.zibaee@gmx.com
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Abstract

A comprehensive study on digestive trypsin was undertaken in the larval midgut of Pieris brassicae L. Results of enzymatic compartmentalization showed a significantly higher activity of crude trypsin in the anterior larval midgut rather than posterior-midgut. Using Diethylaminoethyl cellulose fast flow column chromatography a purified trypsin was obtained by specific activity of 21 U mg−1 protein, recovery of 22%, purification fold of 28-fold and molecular weight of 25 kDa. This purified enzyme showed the highest activity at pH 8 and the corresponding temperature of 40°C. However, the specific inhibitors used including 4-(2-Aminoethyl) benzenesulfonyl fluroride hydrochloride, N-p-Tosyl-L-lysine methyl ester hydrochloride and Soybean Trypsin Inhibitor significantly lowered the activity of the purified enzyme in vitro. Moreover, the activity of trypsin and likewise the nutritional indices were significantly altered in the larval midgut feeding upon the leaves treated by 1 mM concentration of each inhibitor in comparison with control. Determination of enzymatic characteristics of insect trypsins is crucial in paving the path for controlling pests by potential natural compounds via transgenic plants.

Keywords

Digestive trypsincharacterizationenzymatic inhibitionPieris brassicae
Type
Research Papers
Information
Bulletin of Entomological Research , Volume 108 , Issue 4 , August 2018 , pp. 501 - 509
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Copyright
Copyright © Cambridge University Press 2017 

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