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Vol 48(2014) N 6 p. 852-857; DOI 10.1134/S0026893314060132 Full Text

R. Panahi1, E. Vasheghani-Farahani1*, S. A. Shojaosadati1, B. Bambai2

Auto-inducible expression system based on the SigB-dependent ohrB promoter in Bacillus subtilis

1Faculty of Chemical Engineering, Tarbiat Modares University, P.O. Box 14115-114, Tehran, Iran
2National Institute for Genetic Engineering and Biotechnology, Tehran, Iran

*evf@modares.ac.ir
Received - 2014-04-06; Accepted - 2014-05-23

A reliable production of heterologous proteins is important in the field of industrial biotechnology. This can be achieved by applying auto-inducible gene expression systems. Development of a Bacillus subtilis expression plasmid harboring SigB-dependent ohrB promoter was reported. The expression system was subjected to high cell density cultivation to produce xylanase as a stable model protein. The recombinant strain was cultured in a synthetic medium containing glucose as the carbon source. The exponential fed-batch feeding strategy was applied to prevent substrate inhibition. A sharp increase of xylanase activity (about 6-fold) at the end of fermentation was observed as a result of sigma factor B (SigB) protein activation, supporting auto-inducibility of the expression system. For the control strain a specific induction of the xylanase activity was not observed. The recombinant strain showed a 5-fold increase in xylanase activity in comparison with the control strain. In addition, the constructed system displayed the catabolite repression resistance ability. This SigB-dependent expression system can be considered as a biotechnological tool and an alternative to the high costing conventional inducers, e.g. isopropyl-β-galactopyranoside.

auto-inducible expression vector, Bacillus subtilis, fermentation, ohrB promoter, sigma factor B



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