Heterologous expression and enhanced production of β-1,4-glucanase of Bacillus halodurans C-125 in Escherichia coli

Recombinant DNA technology enables us to produce proteins with desired properties and insubstantial amount for industrial applications. Endo-1, 4-β-glucanases (Egl) is one of the major enzyme involved in degradation of cellulose, an important component of plant cell wall. The present study was aimed...

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Published in:	Electronic Journal of Biotechnology Vol. 34; pp. 29 - 36
Main Authors:	Zeeshan, Nadia, Naz, Saher, Naz, Shumaila, Afroz, Amber, Zahur, Muzna, Zia, Safia
Format:	Journal Article
Language:	English
Published:	Elsevier España, S.L.U 01-07-2018 Elsevier
Subjects:	4-β-glucanases Bacillus halodurans Cellulases Cellulose hydrolysis Degradation of cellulose Endo-1 Expression analysis Heterologous expression In silico protein characterization IPTG pET expression system Plant cell wall Expression analysis Endo-1 Plant cell wall Degradation of cellulose Cellulases In silico protein characterization pET expression system IPTG 4-β-glucanases Cellulose hydrolysis Bacillus halodurans Heterologous expression
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Summary:	Recombinant DNA technology enables us to produce proteins with desired properties and insubstantial amount for industrial applications. Endo-1, 4-β-glucanases (Egl) is one of the major enzyme involved in degradation of cellulose, an important component of plant cell wall. The present study was aimed at enhancing the production of endo-1, 4-β-glucanases (Egl) of Bacillus halodurans in Escherichia coli. A putative Egl gene of Bacillus Halodurans was expressed in E. coli by cloning in pET 22b (+). On induction with isopropyl-b-d-1-thiogalactopyranoside, the enzyme expression reached upto ~20% of the cell protein producing 29.2 mg/liter culture. An increase in cell density to 12 in auto-inducing LB medium (absorbance at 600 nm) enhanced β-glucanase production up to 5.4 fold. The molecular mass of the enzyme was determined to be 39 KDa, which is nearly the same as the calculated value. Protein sequence was analyzed by CDD, Pfam, I TASSER, COACH, PROCHECK Servers and putative amino acids involved in the formation of catalytic, substrate and metal binding domains were identified. Phylogenetic analysis of the β-glucanases of B. halodurans was performed and position of Egl among other members of the genus Bacillus producing endo-glucanases was determined. Temperature and pH optima of the enzyme were found to be 60°C and 8.0, respectively, under the assay conditions. Production of endo-1, 4 β-glucanase enzymes from B. halodurans increased several folds when cloned in pET vector and expressed in E. coli. To our knowledge, this is the first report of high-level expression and characterization of an endo-1, 4 β-glucanases from B. halodurans. How to cite: Zeeshan N, Naz S, Naz S et al. Heterologous expression and enhanced production of β-1,4-glucanase of Bacillus halodurans C-125 in E. coli. Electron J Biotechnol 2018;34. https://doi.org/10.1016/j.ejbt.2018.05.001.
ISSN:	0717-3458 0717-3458
DOI:	10.1016/j.ejbt.2018.05.001