Int. J. Mol. Sci. 2011, 12(3), 1767-1786; doi:10.3390/ijms12031767
Article

Importance of the Long-Chain Fatty Acid Beta-Hydroxylating Cytochrome P450 Enzyme YbdT for Lipopeptide Biosynthesis in Bacillus subtilis Strain OKB105

1 Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK 74078, USA 2 Department of Botany and Microbiology, University of Oklahoma, Norman, OK 74078, USA
* Author to whom correspondence should be addressed.
Received: 9 December 2010; in revised form: 8 February 2011 / Accepted: 3 March 2011 / Published: 8 March 2011
(This article belongs to the Special Issue Biosurfactants)
PDF Full-text Download PDF Full-Text [451 KB, uploaded 8 March 2011 09:36 CET]
Abstract: Bacillus species produce extracellular, surface-active lipopeptides such as surfactin that have wide applications in industry and medicine. The steps involved in the synthesis of 3-hydroxyacyl-coenzyme A (CoA) substrates needed for surfactin biosynthesis are not understood. Cell-free extracts of Bacillus subtilis strain OKB105 synthesized lipopeptide biosurfactants in presence of L-amino acids, myristic acid, coenzyme A, ATP, and H2O2, which suggested that 3-hydroxylation occurs prior to CoA ligation of the long chain fatty acids (LCFAs). We hypothesized that YbdT, a cytochrome P450 enzyme known to beta-hydroxylate LCFAs, functions to form 3-hydroxy fatty acids for lipopeptide biosynthesis. An in-frame mutation of ybdT was constructed and the resulting mutant strain (NHY1) produced predominantly non-hydroxylated lipopeptide with diminished biosurfactant and beta-hemolytic activities. Mass spectrometry showed that 95.6% of the fatty acids in the NHY1 biosurfactant were non-hydroxylated compared to only ~61% in the OKB105 biosurfactant. Cell-free extracts of the NHY1 synthesized surfactin containing 3-hydroxymyristic acid from 3-hydroxymyristoyl-CoA at a specific activity similar to that of the wild type (17 ± 2 versus 17.4 ± 6 ng biosurfactant min−1·ng·protein−1, respectively). These results showed that the mutation did not affect any function needed to synthesize surfactin once the 3-hydroxyacyl-CoA substrate was formed and that YbdT functions to supply 3-hydroxy fatty acid for surfactin biosynthesis. The fact that YbdT is a peroxidase could explain why biosurfactant production is rarely observed in anaerobically grown Bacillus species. Manipulation of LCFA specificity of YbdT could provide a new route to produce biosurfactants with activities tailored to specific functions.
Keywords: cytochrome P450; YbdT; long chain fatty acids; beta hydroxylation; Bacillus subtilis; surfactin

Article Statistics

Load and display the download statistics.

Citations to this Article

Cite This Article

MDPI and ACS Style

Youssef, N.H.; Wofford, N.; McInerney, M.J. Importance of the Long-Chain Fatty Acid Beta-Hydroxylating Cytochrome P450 Enzyme YbdT for Lipopeptide Biosynthesis in Bacillus subtilis Strain OKB105. Int. J. Mol. Sci. 2011, 12, 1767-1786.

AMA Style

Youssef NH, Wofford N, McInerney MJ. Importance of the Long-Chain Fatty Acid Beta-Hydroxylating Cytochrome P450 Enzyme YbdT for Lipopeptide Biosynthesis in Bacillus subtilis Strain OKB105. International Journal of Molecular Sciences. 2011; 12(3):1767-1786.

Chicago/Turabian Style

Youssef, Noha H.; Wofford, Neil; McInerney, Michael J. 2011. "Importance of the Long-Chain Fatty Acid Beta-Hydroxylating Cytochrome P450 Enzyme YbdT for Lipopeptide Biosynthesis in Bacillus subtilis Strain OKB105." Int. J. Mol. Sci. 12, no. 3: 1767-1786.

Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert