Characterization of the role of glycine lipids inBacteroides thetaiotaomicron

Author:

Lynch Alli,Tammireddy Seshu R.,Doherty Mary K.,Whitfield Phillip D.,Clarke David J.ORCID

Abstract

AbstractAcylated amino acids function as important components of the cellular membrane in some bacteria. Biosynthesis is initiated by the N-acylation of the amino acid and this is followed by subsequent O-acylation of the acylated molecule resulting in the production of the mature diacylated amino acid lipid. In this study we use both genetics and liquid chromatography-mass spectrometry (LC-MS) to characterize the biosynthesis and function of novel diacylated glycine lipid (GL) species inBacteroides thetaiotaomicron. We, and others, have previously reported the identification of a gene, namedglsBin this study, that encodes a N-acyltransferase activity responsible for the production of a monoacylated glycine called N-acyl-3-hydroxy-palmitoyl glycine (or commendamide). In all of theBacteroidalesgenomes so far sequenced theglsBgene is located immediately downstream from a gene, namedglsA, also predicted to encode a protein with acyltransferase activity. We use LC-MS to show that co-expression ofglsBandglsAresults in the production of GL inEscherichia coli. We constructed a deletion mutant of theglsBgene inB. thetaiotaomicronand we confirm thatglsBis required for the production of GL inB. thetaiotaomicron. Moreover, we show thatglsBis important for the ability ofB. thetaiotaomicronto adapt to stress and colonize the mammalian gut. Therefore, this report is the first to describe the genetic requirements for the biosynthesis of GL, a novel diacylated amino acids species that contributes to fitness in the human gut bacterium,B. thetaiotaomicron.

Publisher

Cold Spring Harbor Laboratory

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