Increasing the stability of sacB transcript improves levansucrase production in Bacillus subtilis

Abstract

Aims: To develop a strategy to increase the stability of transcripts of structural genes expressed under the control of sacR, the leader region of Bacillus subtilis levansucrase gene. Methods and Results: Insertion of Shine Dalgarno like sequences in the 5¢-untranslated sacR region controlling the expression of sacB. Depending on the number of stabilizing sequences inserted and the position of these sequences with respect to the translation start codon, it was observed that the mRNA stability and the final protein production could be increased or decreased. Conclusions: This mRNA stabilization can be used to increase exocellular protein production in the degU32 (Hy) mutant. Significance and Impact of the Study: This approach can be applied to the expression of heterologous genes of biotechnological interest.

Secreted production of Renilla luciferase in Bacillus subtilis

“…Recently, a study has reported a way to improve stability of the endogenous sacB transcript by insertion of Shine-Dalgarno like sequence in the 5 0 -untranslated region. 30 This approach resulted in an enhanced production of extracellular levansucrase in B. subtilis. Along with the present result, it suggests that a strategy by stabilizing the gene transcript might be equally useful for enhanced production of either endogenous or heterologous proteins.…”