Identification of four secondary acyltransferases for lipid A biosynthesis in Vibrio parahaemolyticus.

Abstract

In this study, four genes encoding secondary acyltransferases of lipid A in Vibrio parahaemolyticus ATCC33846 were identified. When the four genes were overexpressed in Escherichia coli MLK1067 that which produces the penta-acylated lipid A lacking the secondary acylation at the C3' position, a C12:0 secondary acyl chain was added at the C3' position of lipid A only in E. coli overexpressing VP_RS01045, but not VP_RS00880, VP_RS08405, or VP_RS12170. When the four genes were overexpressed in E. coli MKV15b that produces lipid IVA , a C12:0 secondary acyl chain was again added at the C3' position in E. coli overexpressing VP_RS01045, but a C14:0 secondary acyl chain was added at the C2' position of lipid A in E. coli overexpressing VP_RS00880, VP_RS08405, or VP_RS12170. The results indicate that four acyltransferases of lipid A are encoded by VP_RS01045, VP_RS00880, VP_RS08405, or VP_RS12170 in V. parahaemolyticus. The acyltransferase encoded by VP_RS01045 adds a C12:0 secondary acyl chain at the C3' position of lipid A, whereas the acyltransferase encoded by VP_RS00880, VP_RS08405, or VP_RS12170 adds a C14:0 secondary acyl chain at the C2' position of lipid A. This work contributes to understanding the biosynthetic pathway of lipid A in V. parahaemolyticus.