Development of a synthetic cumate-inducible gene expression system for Bacillus

A novel inducible gene expression system using p -isopropyl benzoate (cumate) as an inducer was developed for the industrial production hosts, Bacillus subtilis and Bacillus megaterium . Cumate is non-toxic to the host, inexpensive, and carbon source-independent inducer which provides an economical...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2019, Vol.103 (1), p.303-313
Main Authors: Seo, Seung-Oh, Schmidt-Dannert, Claudia
Format: Article
Language:English
Subjects:
Applied Genetics and Molecular Biotechnology
Bacillus
Biomedical and Life Sciences
Biotechnology
Carbon sources
E coli
Escherichia coli
Gene expression
Genetic aspects
Industrial production
Life Sciences
Metabolic engineering
Microbial Genetics and Genomics
Microbiological research
Microbiology
Organic chemistry
Proteins
Pseudomonas putida
Regulatory sequences
Stability
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Summary:A novel inducible gene expression system using p -isopropyl benzoate (cumate) as an inducer was developed for the industrial production hosts, Bacillus subtilis and Bacillus megaterium . Cumate is non-toxic to the host, inexpensive, and carbon source-independent inducer which provides an economical option for large-scale production of valuable proteins and chemicals from Bacillus strains. The synthetic cumate-inducible system was constructed by combining the strong constitutive Bacillus promoter P veg with regulatory elements of the Pseudomonas putida , CymR repressor, and its operator sequence CuO. The designed expression cassette containing a sf GFP reporter under the cumate-inducible promoter was assembled into a Bacillus - E. coli shuttle and gene expression investigated in the two Bacillus strains. Characterization of gene expression levels, expression kinetics, and dose-response to cumate inducer concentration confirmed high-level, but tightly controlled GFP reporter expression in tunable, cumate concentration-dependent manner. Unexpectedly, this expression system works equally well for Escherichia coli , resulting in a platform that can be used both in gram-positive and gram-negative expression host. Its tight regulation and controllable expression makes this system useful for metabolic engineering, synthetic biology studies as well industrial protein production.
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-018-9485-4