title: Synergism of regulatory elements in σ(B)- and σ(A)-dependent promoters enhances recombinant protein expression in Bacillus subtilis.
abstract: 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 σ(B)- and σ(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 σ(B)-promoter was located upstream to σ(A)-promoter but independent to its orientation. A conserved element in either -10 or -35 box of σ(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.
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Phutthamontol, Nakhon Pathom 73170, Thailand.
authors.forename: Teva
authors.initials: T
authors.lastname: Phanaksri
authors.affiliation: Microbial Cell Factory Laboratory, Bioresource Technology Unit, National Center for Genetic Engineering and Biotechnology, Thailand Scinece Park, Pathumthani 12120, Thailand.
authors.forename: Plearnpis
authors.initials: P
authors.lastname: Luxananil
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Phutthamontol, Nakhon Pathom 73170, Thailand; Department of Biochemistry, Faculty of Science, Mahidol University, Phayathai, Bangkok 10400, Thailand.
authors.forename: Sakol
authors.initials: S
authors.lastname: Panyim
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Phutthamontol, Nakhon Pathom 73170, Thailand. Electronic address: witoon.tir@mahidol.ac.th.
authors.forename: Witoon
authors.initials: W
authors.lastname: Tirasophon
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics*; D001483:Base Sequence; D017124:Conserved Sequence / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics*; D012808:Sigma Factor / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D012808:Sigma Factor
keywords: Bacillus subtilis; Double promoter; Green fluorescent protein; Recombinant protein expression; Synergism; σ(A)-dependent promoter; σ(B)-dependent promoter