Efficient Protein Expression and Biosynthetic Gene Cluster Regulation in Bacillus subtilis Driven by a T7-BOOST System.
Yaokang WuYang LiYuting ZhangYanfeng LiuJianghua LiGuocheng DuXueqin LvLong LiuPublished in: ACS synthetic biology (2023)
Bacillus subtilis is a generally recognized as safe microorganism that is widely used for protein expression and chemical production, but has a limited number of genetic regulatory components compared with the Gram-negative model microorganism Escherichia coli . In this study, a two-module plug-and-play T7- b ased o ptimized o utput s trategy for t ranscription (T7-BOOST) systems with low leakage expression and a wide dynamic range was constructed based on the inducible promoters P hy-spank and P xylA . The first T7 RNA polymerase-driven module was seamlessly integrated into the genome based on the CRISPR/Cpf1 system, while the second expression control module was introduced into low, medium, and high copy plasmids for characterization. As a proof of concept, the T7-BOOST systems were successfully employed for whole-cell catalysis production of γ-aminobutyric acid (109.8 g/L with a 98.0% conversion rate), expression of human α S1 casein and human lactoferrin, and regulation of exogenous lycopene biosynthetic gene cluster and endogenous riboflavin biosynthetic gene cluster. Overall, the T7-BOOST system serves as a stringent, controllable, and effective tool for regulating gene expression in B. subtilis .
Keyphrases
- bacillus subtilis
- genome wide
- poor prognosis
- escherichia coli
- gene expression
- gram negative
- copy number
- endothelial cells
- dna methylation
- multidrug resistant
- genome editing
- crispr cas
- binding protein
- induced pluripotent stem cells
- genome wide identification
- stem cells
- pluripotent stem cells
- pseudomonas aeruginosa
- cell therapy
- staphylococcus aureus
- mesenchymal stem cells
- bone marrow
- biofilm formation
- recombinant human
- candida albicans