Jie Yue1,2,3, Gang Fu2,3, Dawei Zhang4,5, Jianping Wen1. 1. Department of Biological Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China. 2. Tianjin Institutes of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People's Republic of China. 3. Key Laboratories of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People's Republic of China. 4. Tianjin Institutes of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People's Republic of China. zhang_dw@tib.cas.cn. 5. Key Laboratories of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People's Republic of China. zhang_dw@tib.cas.cn.
Abstract
OBJECTIVES: To improve heterologous proteins production, we constructed a maltose-inducible expression system in Bacillus subtilis. RESULTS: An expression system based on the promoter for maltose utilization constructed in B. subtilis. Successively, to improve the performance of the P malA -derived system, mutagenesis was employed by gradually shortening the length of P malA promoter and altering the spacing between the predicted MalR binding site and the -35 region. Furthermore, deletion of the maltose utilization genes (malL and yvdK) improved the P malA promoter activity. Finally, using this efficient maltose-inducible expression system, we enhanced the production of luciferase and D-aminoacylase, compared with the P hpaII system. CONCLUSIONS: A maltose-inducible expression system was constructed and evaluated. It could be used for high level expression of heterologous proteins production.
OBJECTIVES: To improve heterologous proteins production, we constructed a maltose-inducible expression system in Bacillus subtilis. RESULTS: An expression system based on the promoter for maltose utilization constructed in B. subtilis. Successively, to improve the performance of the P malA -derived system, mutagenesis was employed by gradually shortening the length of P malA promoter and altering the spacing between the predicted MalR binding site and the -35 region. Furthermore, deletion of the maltose utilization genes (malL and yvdK) improved the P malA promoter activity. Finally, using this efficient maltose-inducible expression system, we enhanced the production of luciferase and D-aminoacylase, compared with the P hpaII system. CONCLUSIONS: A maltose-inducible expression system was constructed and evaluated. It could be used for high level expression of heterologous proteins production.
Entities:
Keywords:
Bacillus subtilis; D-Aminoacylase; Expression vector; Gene deletion; Luciferase; P malA promoter