Synergism of regulatory elements in σB- and σA-dependent promoters enhances recombinant protein expression in Bacillus subtilis

Strong promoter is an essential factor for production of recombinant protein in various expression systems including Bacillus subtilis. In this study, we described a strategy to improve the expression efficiency using synthetic double promoter. Assembly of the conserved elements from sigma(B)- and sigma(A)-dependent promoters constitutively improved the yield of recombinant protein approximately 2-3-fold in both exponential and stationary growth phase. The synergistic effect in the double promoter was observed only when sigma(B)- promoter was located upstream to sigma(A)-promoter but independent to its orientation. A conserved element in either -10 or -35 box of sigma(B)-promoter is sufficient to promote the synergism. Hence, this simple strategy of promoter engineering could be an effective way to generate a pool of strong constitutive promoters applicable for heterologous protein expression in B. subtilis in the future. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.