Glucose-1-phosphate thymidylyltransferase promotes the production of 3-O-α-mycarosylerythronolide B in Streptomyces coelicolor

J Appl Microbiol. 2024 Dec 2;135(12):lxae291. doi: 10.1093/jambio/lxae291.

Abstract

Aims: The main objective of this study was to produce erythronolide B (EB) and 3-O-α-mycarosylerythronolide B (MEB) in Streptomyces coelicolor and enhance the MEB production by expressing the glucose-1-phosphate thymidylyltransferase (RfbA).

Methods and results: We expressed eryF and eryB genes (eryBII, eryBIII, eryBIV, eryBV, eryBVI, and eryBVII) to produce EB and MEB. The expression was confirmed by quantitative real-time polymerase chain reaction. Furthermore, the MEB's production was improved by more than 100-fold by expressing an enzyme, RfbA, which is absent from the erythromycin gene cluster, to promote the biosynthesis of TDP-L-mycarose. We discuss the feasibility of alternative Streptomyces species for erythromycin production based on the presence or absence of RfbA.

Conclusions: The RbfA enzyme from Saccharopolyspora erythraea was expressed in S. coelicolor M1152 along with the MEB biosynthesis pathway, resulting in a large increase in MEB production (>100-fold).

Keywords: Streptomyces coelicolor; 3-O-α-mycarosylerythronolide B; erythromycin; glucose-1-phosphate thymidylyltransferase; heterologous expression.

MeSH terms

  • Anti-Bacterial Agents / biosynthesis
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Erythromycin* / analogs & derivatives
  • Nucleotidyltransferases / genetics
  • Nucleotidyltransferases / metabolism
  • Saccharopolyspora / enzymology
  • Saccharopolyspora / genetics
  • Saccharopolyspora / metabolism
  • Streptomyces coelicolor* / enzymology
  • Streptomyces coelicolor* / genetics
  • Streptomyces coelicolor* / metabolism

Substances

  • Erythromycin
  • glucose-1-phosphate thymidylyltransferase
  • Nucleotidyltransferases
  • Bacterial Proteins
  • Anti-Bacterial Agents