A facile and robust T7-promoter-based high-expression of heterologous proteins in Bacillus subtilis
نویسندگان
چکیده
Abstract To mimic the Escherichia coli T7 protein expression system, we developed a facile promoter-based system in an industrial microorganism Bacillus subtilis . This has two parts: new B. strain SCK22 and plasmid pHT7. construct SCK22, RNA polymerase gene was inserted into chromosome, several genes, such as major protease spore generation-related gene, fermentation foam were knocked out to facilitate good high-density cell fermentation. The of target can be subcloned pHT7, where under tight control promoter with ribosome binding site (RBS) sequence (i.e., AAGGAGG). A few recombinant proteins green fluorescent protein, ?-glucan phosphorylase, inositol monophosphatase, phosphoglucomutase, 4-?-glucanotransferase) expressed approximately 25–40% levels relative cellular total estimated by SDS-PAGE using SCK22/pHT7-derived plasmid. fed-batch high-cell density conducted 5-L fermenter, producing up 4.78 g/L monophosphatase. advantageous features, as, wide applicability for proteins, high level, easy genetic operation, transformation efficiency, stability, suitability Graphical
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ژورنال
عنوان ژورنال: Bioresources and Bioprocessing
سال: 2022
ISSN: ['2197-4365']
DOI: https://doi.org/10.1186/s40643-022-00540-4