cDNA cloning, heterologous expression and characterization of a cell wall invertase from copper tolerant population of Elsholtzia haichowensis
Abstract
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◦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 Km and Vmax values of FCWI for hydrolyzing sucrose were 0.282 mM and 1.576 μkat/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
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Artikel in diesem Heft
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Artikel in diesem Heft
- Down into the Earth: microbial diversity of the deepest cave of the world
- Factors influencing synergistic antimicrobial activity of thymol and nisin against Shigella spp. in sugarcane juice
- Effects of low-temperature hardening on the biochemical response of winter oilseed rape seedlings inoculated with the spores of Leptosphaeria maculans
- Mitochondrial structures during seed germination and early seedling development in Arabidopsis thaliana
- Transcriptome analysis for identification of indigo biosynthesis pathway genes in Polygonum tinctorium
- Different components of plant diversity suggest the protection of a large area for the conservation of a riparian ecosystem
- Anatomical adaptations of the desert species Stipa lagascae against drought stress
- Effects of ammonium ion on cell growth and biosynthesis of shikonin derivatives in callus tissues of Arnebia euchroma
- cDNA cloning, heterologous expression and characterization of a cell wall invertase from copper tolerant population of Elsholtzia haichowensis
- Construction of cDNA library from Prunus campanulata leaves and preliminary expressed sequence tag (EST) analysis during cold stress
- Phylogenetic utility of the geometric model of the body form in leeches (Clitellata: Hirudinida)
- Substrate choice by the alien snail Ferrissia fragilis (Gastropoda: Planorbidae) in an industrial area: A case study in a forest pond (Southern Poland)
- History of two critically endangered grassland snails (Pulmonata: Helicellinae) in the Czech Republic with first molecular data on extinct populations
- Monteustium marezensis gen. n., sp. n. and the first record of Italustiun eframi (Acari: Prostigmata: Erythraeidae: Balaustiinae) from Montenegro
- Lithobius (Ezembius) laevidentata sp. n., a new species (Chilopoda: Lithobiomorpha: Lithobiidae) from the Northwest region of China
- Aphids in jeopardy? Aphid communities on xerothermic habitats
- Habitat and weather requirements of diurnal raptors wintering in river valleys