RRC ID 53782
著者 Ito T, Yamauchi A, Hemmi H, Yoshimura T.
タイトル Ophthalmic acid accumulation in an Escherichia coli mutant lacking the conserved pyridoxal 5'-phosphate-binding protein YggS.
ジャーナル J Biosci Bioeng
Abstract Escherichia coli YggS is a highly conserved pyridoxal 5'-phosphate (PLP)-binding protein whose biochemical function is currently unknown. A previous study with a yggS-deficient E. coli strain (ΔyggS) demonstrated that YggS controls l-Ile- and l-Val-metabolism by modulating 2-ketobutyrate (2-KB), l-2-aminobutyrate (l-2-AB), and/or coenzyme A (CoA) availability in a PLP-dependent fashion. In this study, we found that ΔyggS accumulates an unknown metabolite as judged by amino acid analyses. LC/MS and MS/MS analyses of the compound with propyl chloroformate derivatization, and co-chromatography analysis identified this compound as γ-l-glutamyl-l-2-aminobutyryl-glycine (ophthalmic acid), a glutathione (GSH) analogue in which the l-Cys moiety is replaced by l-2-AB. We also determine the metabolic consequence of the yggS mutation. Absence of YggS initially increases l-2-AB availability, and then causes ophthalmic acid accumulation and CoA limitation in the cell. The expression of a γ-glutamylcysteine synthetase and a glutathione synthetase in a ΔyggS background causes high-level accumulation of ophthalmic acid in the cells (∼1.2 nmol/mg cells) in a minimal synthetic medium. This opens the possibility of a first fermentative production of ophthalmic acid.
巻・号 122(6)
ページ 689-693
公開日 2016-12-1
DOI 10.1016/j.jbiosc.2016.06.010
PII S1389-1723(16)30145-1
PMID 27426274
MeSH Carrier Proteins / genetics* Carrier Proteins / metabolism Coenzyme A / metabolism Conserved Sequence Dipeptides / genetics Dipeptides / metabolism Escherichia coli / genetics* Escherichia coli / metabolism* Escherichia coli Proteins / genetics* Escherichia coli Proteins / metabolism Gene Deletion* Glutamate-Cysteine Ligase / genetics Glutamate-Cysteine Ligase / metabolism Glutathione Synthase / genetics Glutathione Synthase / metabolism Oligopeptides / biosynthesis* Organisms, Genetically Modified Phosphate-Binding Proteins / genetics Phosphate-Binding Proteins / metabolism Tandem Mass Spectrometry
IF 2.366
引用数 11
リソース情報
原核生物(大腸菌)