文献类型：期刊文献

英文题名：Purification and biochemical characterization of a novel transglutaminase from Mythimna separata larvae (Noctuidae, Lepidoptera)

作者：Zhang, Lei[1];Rao, Wenbing[1];Muhayimana, Solange[1];Zhang, Xianfei[1];Xu, Jiuyong[1];Xiao, Ciying[2];Huang, Qingchun[1]

机构：[1]East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab Chem Biol, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2018

卷号：265

起止页码：1

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20174504364755);WOS:【SCI-EXPANDED(收录号:WOS:000418596500001)】；

基金：We gratefully acknowledge financial support from Shanghai Foundation of Science and Technology (No. 15431906800).

语种：英文

外文关键词：Larval transglutaminase; Mythimna separata; Purification; Biochemical property

摘要：A novel transglutaminase (MsTGase) from Mythimna separata larvae was separated and purified; its biochemical property and enzymatic catalytic activities were investigated. MsTGase was obtained chromatographically by the precipitation of Sephadex G-100 gel and DEAE-Cellulose-52 ion-exchange column with 48-fold purification and a reproducible yield of approximately 12%. Molecular weight of the MsTGase was 63.5 KDa and its N-terminal amino acid sequence was GKIEEG-LVI. Michaelis constant of the MsTGase for the substrate N-CBZ-GlnGly was 12.83 mM with a V-max of 7.99 U/mL. Optimum conditions for MsTGase activity were at 42 degrees C and pH7.5. The enzyme didn't possess metal ion at its catalytic active site; its activity could be significantly inhibited by Mg2+, but activated by Ca2+. Chlorpyrifos and spinosad showed a strong potential to increase MsTGase activity, supporting the view that MsTGase was a novel target. Moreover, the formation of intermolecular crosslinks of casein and bovine serum albumin polymerized by MsTGase in the presence of DTT was observed. These findings pave the way for future studies on the physiological role of MsTGase and the potential impact of its regulation on MsTGase-associated pest management.