Cysteine biosynthesis in Lactobacillus casei: identification and characterization of a serine acetyltransferase

FEMS Microbiology Letters
Biljana BogicevicStefan Irmler

Abstract

In bacteria, cysteine can be synthesized from serine by two steps involving an L-serine O-acetyltransferase (SAT) and a cysteine synthase (CysK). While CysK is found in the publicly available annotated genome from Lactobacillus casei ATCC 334, a gene encoding SAT (cysE) is missing. In this study, we found that various strains of L. casei grew in a chemically defined medium containing sulfide as the sole sulfur source, indicating the presence of a serine O-acetyltransferase. The gene lying upstream of cysK is predicted to encode a homoserine trans-succinylase (metA). To study the function of this gene, it was cloned from L. casei FAM18110. The purified, recombinant protein did not acylate L-homoserine in vitro. Instead, it catalyzed the formation of O-acetyl serine from L-serine and acetyl-CoA. Furthermore, the plasmid expressing the L. casei gene complemented an Escherichia coli cysE mutant strain but not an E. coli metA mutant. This clearly demonstrated that the gene annotated as metA in fact encodes the SAT function and should be annotated as cysE.

References

Apr 15, 2004·Current Opinion in Biotechnology·Roland J SiezenBas Teusink
Sep 30, 2004·Journal of Dairy Science·H M FarrellH E Swaisgood
Dec 8, 2004·Archives of Biochemistry and Biophysics·Corey M JohnsonPaul F Cook
Jan 6, 2005·Journal of Biochemistry·Shin-ichiro KobayashiHiroshi Takagi
Oct 13, 2006·Proceedings of the National Academy of Sciences of the United States of America·K MakarovaD Mills
Nov 13, 2007·Applied and Environmental Microbiology·Stefan IrmlerHélène Berthoud
Dec 8, 2007·Applied Microbiology and Biotechnology·Sophie LandaudPascal Bonnarme
Jun 10, 2008·Applied and Environmental Microbiology·Mengjin LiuRoland J Siezen
Nov 25, 2011·Applied Microbiology and Biotechnology·Biljana BogicevicStefan Irmler

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Datasets Mentioned

BETA
KU216159

Methods Mentioned

BETA
acylation
Protein
electrophoresis
PCR
enzymatic assay
acetylation

Software Mentioned

Data Analysis
InterProScan

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