2024-03-28T17:21:06Z
https://nagoya.repo.nii.ac.jp/oai
oai:nagoya.repo.nii.ac.jp:00023609
2023-01-16T04:12:59Z
643:666:667
Ophthalmic acid accumulation in an Escherichia coli mutant lacking the conserved pyridoxal 5′-phosphate-binding protein YggS
Ito, Tomokazu
70138
Yamauchi, Ayako
70139
Hemmi, Hisashi
70140
Yoshimura, Tohru
70141
Pyridoxal 5′-phosphate
Ophthalmic acid
YggS
Escherichia coli
Glutathione
Amino acid
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.
journal article
Elsevier
2016-12
application/pdf
application/pdf
Journal of Bioscience and Bioengineering
6
122
689
693
http://dx.doi.org/10.1016/j.jbiosc.2016.06.010
http://hdl.handle.net/2237/25806
1389-1723
https://nagoya.repo.nii.ac.jp/record/23609/files/yggs2_revise.pdf
https://nagoya.repo.nii.ac.jp/record/23609/files/YggS_second-revise.pdf
eng
https://doi.org/10.1016/j.jbiosc.2016.06.010
© 2016. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/