Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp. G1 in E. coli.

Journal of Industrial Microbiology & Biotechnology
Rui Min OngRosli Md Illias

Abstract

The cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) gene from Bacillus sp. G1 was successfully isolated and cloned into Escherichia coli. Analysis of the nucleotide sequence revealed the presence of an open reading frame of 2,109 bp and encoded a 674 amino acid protein. Purified CGTase exhibited a molecular weight of 75 kDa and had optimum activity at pH 6 and 60 degrees C. Heterologous recombinant protein expression in E. coli is commonly problematic causing intracellular localization and formation of inactive inclusion bodies. This paper shows that the majority of CGTase was secreted into the medium due to the signal peptide of Bacillus sp. G1 that also works well in E. coli, leading to easier purification steps. When reacted with starch, CGTase G1 produced 90% beta-cyclodextrin (CD) and 10% gamma-CD. This enzyme also preferred the economical tapioca starch as a substrate, based on kinetics studies. Therefore, CGTase G1 could potentially serve as an industrial enzyme for the production of beta-CD.

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Citations

Sep 19, 2009·Applied Microbiology and Biotechnology·Hans LeemhuisLubbert Dijkhuizen
Jan 18, 2013·Applied Microbiology and Biotechnology·Kian Mau GohKok-Gan Chan
Aug 18, 2012·Journal of Industrial Microbiology & Biotechnology·Hua LiuJian Chen
Sep 7, 2011·Applied Biochemistry and Biotechnology·Tsvetina KitayskaAlexandra Ivanova Tonkova
May 29, 2012·Applied Biochemistry and Biotechnology·Berhane Teklehaimanot TesfaiJing Wu
Jul 4, 2012·International Journal of Molecular Sciences·Poh Hong GohKian Mau Goh
Sep 6, 2012·International Journal of Molecular Sciences·Abdelnasser S S IbrahimGarabed Antranikian
Nov 5, 2014·Bioprocess and Biosystems Engineering·Ashraf F ElbazAhmed ElMekawy
Mar 30, 2012·Journal of Molecular Microbiology and Biotechnology·Mohd Anuar JonetRosli Md Illias
Jan 9, 2013·Journal of Molecular Microbiology and Biotechnology·Menaga SubramaniamAbdul Rahim Raha
Apr 15, 2018·Applied and Environmental Microbiology·Thomas BaumgartenJan-Willem de Gier
Feb 21, 2021·Journal of the Science of Food and Agriculture·Qing MengTao Zhang
Sep 28, 2012·Journal of Agricultural and Food Chemistry·Jingwen ZhouJian Chen
Aug 28, 2018·Protein Expression and Purification·Fabiane Cristina Dos Santos, Ione Parra Barbosa-Tessmann

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