Heterologous production of paromamine in Streptomyces lividans TK24 using kanamycin biosynthetic genes from Streptomyces kanamyceticus ATCC12853

Mol Cells. 2009 May 31;27(5):601-8. doi: 10.1007/s10059-009-0080-5. Epub 2009 May 15.

Abstract

The 2-deoxystreptamine and paromamine are two key intermediates in kanamycin biosynthesis. In the present study, pSK-2 and pSK-7 recombinant plasmids were constructed with two combinations of genes: kanABK and kanABKF and kacA respectively from kanamycin producer Streptomyces kanamyceticus ATCC12853. These plasmids were heterologously expressed into Streptomyces lividans TK24 independently and generated two recombinant strains named S. lividans Sk-2/SL and S. lividans SK-7/SL, respectively. ESI/ MS and ESI-LC/MS analysis of the metabolite from S. lividans SK-2/SL showed that the compound had a molecular mass of 163 [M + H]+, which corresponds to that of 2-deoxystreptamine. ESI/MS and MS/MS analysis of metabolites from S. lividans SK-7/SL demonstrated the production of paromamine with a molecular mass of 324 [M + H]+. In this study, we report the production of paromamine in a heterologous host for the first time. This study will evoke to explore complete biosynthetic pathways of kanamycin and related aminoglycoside antibiotics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminoglycosides / genetics
  • Aminoglycosides / metabolism*
  • Anti-Bacterial Agents / biosynthesis*
  • Anti-Bacterial Agents / chemistry
  • Biotechnology
  • Genetic Engineering
  • Hexosamines / genetics
  • Hexosamines / metabolism
  • In Vitro Techniques
  • Kanamycin / biosynthesis*
  • Kanamycin / chemistry
  • Mass Spectrometry
  • Molecular Weight
  • Species Specificity
  • Streptomyces lividans / genetics*
  • Streptomyces lividans / growth & development
  • Streptomyces lividans / metabolism*

Substances

  • Aminoglycosides
  • Anti-Bacterial Agents
  • Hexosamines
  • paromamine
  • Kanamycin
  • 2-deoxystreptamine