Characterization of hydantoin-5-propionic acid amidohydrolase involved in ergothioneine utilization in Burkholderia sp. HME13

Biosci Biotechnol Biochem. 2023 Mar 21;87(4):411-419. doi: 10.1093/bbb/zbad002.

Abstract

In our previous study, ertABC genes encoding ergothionase, thiourocanate hydratase, and 3-(5-oxo-2-thioxoimidazolidin-4-yl) propionic acid desulfhydrase were identified, all of which may be involved in ergothioneine utilization of Burkholderia sp. HME13. In this study, we identify the ertD gene encoding metal-dependent hydantoin-5-propionic acid amidohydrolase in this strain. Mn2+-containing ErtD showed maximum activity at 45 °C and pH 8.5 and was stable at temperatures up to 45 °C. The Km and Vmax values of Mn2+-containing ErtD for hydantoin-5-propionic acid were 2.8 m m and 16 U/mg, respectively. Real-time polymerase chain reaction (PCR) revealed that ertD expression levels in Burkholderia sp. HME13 cells cultivated in ergothioneine medium were 3.3-fold higher than those in cells cultivated in Luria-Bertani (LB) medium. ErtD activity in the crude extract from Burkholderia sp. HME13 cells cultured in ergothioneine medium was 0.018 U/mg, whereas that in LB medium was not detected. Accordingly, we suggest that ErtD is involved in ergothioneine utilization in this strain.

Keywords: Burkholderia; ergothioneine; hydantoin-5-propionic acid; hydantoin-5-propionic acid amidohydrolase.

MeSH terms

  • Amidohydrolases / metabolism
  • Burkholderia* / genetics
  • Burkholderia* / metabolism
  • Ergothioneine*
  • Hydantoins* / metabolism

Substances

  • Ergothioneine
  • hydantoin-5-propionic acid
  • Amidohydrolases
  • Hydantoins