Cloning and expression of the xynA1 gene encoding a xylanase of the GH10 group in Caulobacter crescentus

Applied Biochemistry and Biotechnology
Luciana GracianoRita de Cássia Garcia Simão

Abstract

Caulobacter crescentus (NA1000 strain) are aquatic bacteria that can live in environments of low nutritional quality and present numerous genes that encode enzymes involved in plant cell wall deconstruction, including five genes for β-xylosidases (xynB1-xynB5) and three genes for xylanases (xynA1-xynA3). The overall activity of xylanases in the presence of different agro-industrial residues was evaluated, and it was found that the residues from the processing of corn were the most efficient in inducing bacterial xylanases. The xynA1 gene (CCNA_02894) encoding a predicted xylanase of group 10 of glyco-hydrolases (GH10) that was efficiently overexpressed in Escherichia coli LMG194 using 0.02 % arabinose, after cloning into the vector pJet1.2blunt and subcloning into the expression vector pBAD/gIII, provided a fusion protein that contained carboxy-terminal His-tags, named XynA1. The characterization of pure XynA1 showed an enzymatic activity of 18.26 U mL(-1) and a specific activity of 2.22 U mg-(1) in the presence of xylan from beechwood as a substrate. XynA1 activity was inhibited by EDTA and metal ions such as Cu(2+) and Mg(2+). By contrast, β-mercaptoethanol, dithiothreitol (DTT), and Ca(2+) induced recombinant enzyme activity...Continue Reading

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Jul 19, 2012·World Journal of Microbiology & Biotechnology·Luciana GracianoRita de Cássia Garcia Simão
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Oct 12, 2012·Applied Biochemistry and Biotechnology·Juliana Moço CorrêaRita de Cássia Garcia Simão
Oct 22, 2013·Applied Biochemistry and Biotechnology·Juliana Moço CorrêaRita de Cássia Garcia Simão
Mar 1, 1993·World Journal of Microbiology & Biotechnology·S NakamuraK Horikoshi

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Citations

Nov 6, 2015·Applied Biochemistry and Biotechnology·Adilson BosettoRita de Cássia Garcia Simão
Mar 6, 2017·Microbial Cell Factories·Laia GifreElena Garcia-Fruitós
Jun 27, 2019·Applied Biochemistry and Biotechnology·Attia Hamid, Muhammad Nauman Aftab
Aug 31, 2021·Applied Microbiology and Biotechnology·Johan S HeroM Alejandra Martinez

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