Isoprenoids are produced via the mevalonic acid (MVA) and methylerythritol phosphate (MEP) pathways and are among the most valuable secondary metabolites. Isoprenoids with medicinal value have proven extremely difficult to synthesize and microbial production has been complicated due to cell death as a result of the buildup of toxic precursors in engineered cells. Bacillus subtilis exhibits a greater tolerance towards these toxic precursors than E. coli, possibly due to the presence of the yhfR gene, whose enzymatic product is thought to be responsible the detoxification of pathway intermediates. We replaced the yhfR gene with a spectinomycin resistance cassette, confirmed by qPCR and sequencing, and have analyzed methylbutanol and isoprene production in engineered and wild-type cells. Our research aims to determine whether the enzyme coded for by yhfR plays a role in the conversion of toxic precursors to volatile compounds that can be released into the environment.

Deletion of the yhfR gene in Bacillus subtilis and its effects on isoprene and methylbutanol production

245th National Meeting (2013)

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