PksS from Bacillus subtilis is a cytochrome P450 involved in bacillaene metabolism

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
Stephanie Anne Antolak (Creator)
Gregory M. Raner, Associate Professor and Graduate Director (Creator)
Jason J. Reddick, Associate Professor (Creator)
Institution
The University of North Carolina at Greensboro (UNCG )
Web Site: http://library.uncg.edu/

Abstract: As part of the pksX gene cluster of Bacillus subtilis strain 168, pksX has been preliminarily annotated as a cytochrome P450 homolog that hydroxylates the polyketide product of this cluster, which was recently shown to be involved in the biosynthesis of bacillaene and dihydrobacillaene. Here we report that there is a frame-shift error in the reported sequence for pksX, and that we have successfully cloned, overexpressed, and purified the protein encoded by the corrected sequence. By utilizing electronic absorption spectrophotometry, we have observed that the ferrous CO complex of pksX absorbs maximally near 450 nm, which confirms the annotation that this protein is a cytochrome P450. We have also established a cell-free system derived from crude cytosolic B. subtilis protein extracts which provides reductase activity essential to sustaining the putative catalytic cycle of pksX. Using LC–MS analysis we have collected data which suggests that the substrate for pksX is dihydrobacillaene.

Additional Information

Publication
Biochemical and Biophysical Research Communications 358:363-367 (2007).
Language: English
Date: 2007
Keywords
Polyketide biosynthesis, Bacillus subtilis, Bacillaene, Cytochrome P450