标題：cDNA cloning, heterologous expression and characterization of a cell wall invertase from copper tolerant population of Elsholtzia haichowensis作者：Liu, Chen; Xu, Zhongrui; Cai, Shenwen; Zhang, Luan; Xiong, Zhiting

來源出版物：BIOLOGIA 卷：70 期：8 頁：1063-1069 DOI：10.1515/biolog-2015-0120 出版年：AUG 2015

摘要：The main objective of the present study was to clone, heterologously express and characterize a novel cell wall invertase (FCWI) from a Cu tolerant population of Elsholtzia haichowensis. The full-length FCWI cDNA contained an open reading frame (ORF) of 1671 bp which encoded a 556-amino-acid protein. The theoretical molecular mass and pI of the deduced protein were 62.5 kDa and 9.29, respectively. Phylogenetic analysis showed that FCWI had a closer evolutionary relationship to cell wall invertase of dicot. FCWI was expressed in methylotrophic yeast Pichia pastoris and purified to near homogeneity. Recombinant FCWI enzyme had pH optima of 4.0 and temperature optima of 50 degrees C. Activity analyses in the presence of various metal cations indicated that FCWI was completely inhibited by Hg2+ (0%), while retained 77.4% activity when exposure to Cu2+. The K-m and V-max values of FCWI for hydrolyzing sucrose were 0.282 mM and 1.576 mu kat/mg, respectively. This is the first report that the heterologous expression and characterization of a cell wall invertase from a Cu tolerant population of E. haichowensis. These results helped to understanding the characteristics of FCWI and its physiological role in the resistance mechanisms of Cu tolerant plants.

入藏号：WOS:000361772300009

文獻類型：Article

語種：English

作者關鍵詞：cell wall invertase, copper, enzyme kinetics, Elsholtzia haichowensis, heterologous expression, Pichia pastoris

擴展關鍵詞：PLANT INVERTASES; EXTRACELLULAR INVERTASE; KUMMEROWIA-STIPULACEA; ROOT-GROWTH; STRESS; ROLES; IDENTIFICATION; METABOLISM; ISOZYMES; PROTEIN

通訊作者地址：Liu, Chen; Wuhan Univ, Sch Resource & Environm Sci, Wuhan 430079, Hubei, Peoples R China.

電子郵件地址：chenliu@whu.edu.cn

地址：

[Liu, Chen; Xu, Zhongrui; Xiong, Zhiting] Wuhan Univ, Sch Resource & Environm Sci, Wuhan 430079, Hubei, Peoples R China.

[Cai, Shenwen] Zunyi Normal Coll, Coll Resource & Environm, Zunyi 563002, Guizhou, Peoples R China.

[Zhang, Luan] Fujian Agr & Forestry Univ, Sch Resource & Environm Sci, Fuzhou 350002, Fujian, Peoples R China.

研究方向：Life Sciences & Biomedicine - Other Topics

ISSN：0006-3088

eISSN：1336-9563

影響因子（2014）：0.827