Random knock-in expression system for high yield production of heterologous protein in Bacillus subtilis

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Random knock-in expression system for high yield production of heterologous protein in Bacillus subtilis
Da-Eun Jeong; Younju So; S Y Park; Seung-Hwan Park; Soo Keun Choi
Bibliographic Citation
Journal of Biotechnology, vol. 266, pp. 50-58
Publication Year
Chromosome-integrated recombinant protein expression in bacteria has advantages for the stable maintenance of genes without any use of antibiotics during large-scale fermentation. Even though different levels of gene expression were reported, depending upon their chromosomal position in bacterial species, only a limited number of integration sites have been used in B. subtilis. In this study, we randomly integrated the GFP and AprE expression cassettes into the B. subtilis genome to determine integration sites that can produce a high yield of heterologous protein expression. Our mariner transposon-based expression cassette integration system was able to find integration sites, which can produce up to 2.9-fold and 1.5-fold increased expression of intracellular GFP and extracellular AprE, respectively, compared to the common integration site amyE. By analyzing the location of integration sites, we observed an adjacent promoter effect, gene dosage effect, and gene knock-out effect all complexly contributing to the increased level of integrated gene expression. Besides obtaining a high yield of heterologous protein expression, our system can also provide a wide-range of expression to expand the systematic application for steady-state metabolic protein production. ⓒ 2017 Elsevier B.V.
Bacillus expression systemChromosomal integration siteMariner-based transposonRandom knock-in
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Division of Research on National Challenges > Infectious Disease Research Center > 1. Journal Articles
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