Cytotoxicity related to flux through each of the isoprenoid pathways has been attributed to buildup of prenyl diphosphate isoprenoid precursors, including dimethyllallyl diphosphate (DMAPP). Isoprene is enzymatically produced in some organisms from DMAPP; we hypothesize that it is produced as a safety valve metabolite to rid the cells of toxic prenyl diphosphates.  To explore cytotoxicity and isoprene production related to the endogenous methylerythritol phosphate (MEP) pathway in Bacillus subtilis, we are growing cells with permeable prenyl alcohols and engineering a nudF gene knockout. The prenyl alcohols cause dose-dependent MEP-related cytotoxicity when fed to cells, while the nudF enzyme product consumes DMAPP to produce prenyl alcohol. We measure uptake and production of these alcohols using a novel GS-MS metthod, and we also monitor isoprene levels and viability in treated cells. These approaches allow us to investigate the role of isoprene in relieving MEP pathway toxicity in B. subtilis.

Exploring the role of prenyl alcohols in isoprene biosynthesis and isoprenoid pathway-related cytotoxicity in Bacillus subtilis

243rd National Meeting (2012)

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