Evaluation of heterologous promoters for genetic analysis of Actinoplanes teichomyceticus--Producer of teicoplanin, drug of last defense

J Biotechnol. 2013 Dec;168(4):367-72. doi: 10.1016/j.jbiotec.2013.10.018. Epub 2013 Oct 24.

Abstract

Actinoplanes teichomyceticus is the only known producer of the valuable glycopeptide antibiotic teicoplanin. Random mutagenesis and selection were extensively applied to teicoplanin producers, while the gene engineering methods were not used, because of the paucity of genetic tools for A. teichomyceticus. Particularly, availability of promoters of different strength that are functional in Actinoplanes would be very useful for overexpression of beneficial genes. Here we report the use of a glucuronidase reporter system (gusA) for studying transcriptional activity in A. teichomyceticus and describe the behavior of a set of heterologous promoters in this strain. We reveal several elements that exceed in their strength the well-established Streptomyces promoter ermEp, underscoring the utility of the gusA reporter for Actinoplanes sp. Remarkable overproduction of teicoplanin was achieved by constructing strains carrying additional copies of the regulatory gene tcp28 under the control of one of the two most active promoters, moeE5p and actp, discovered in this study.

Keywords: Actinoplanes teichomyceticus; Overexpression; Promoter; Reporter gene; Teicoplanin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / biosynthesis*
  • Anti-Bacterial Agents / metabolism
  • Anti-Bacterial Agents / therapeutic use
  • Genes, Reporter
  • Glucuronidase / genetics
  • Micromonosporaceae / genetics*
  • Micromonosporaceae / growth & development
  • Promoter Regions, Genetic*
  • Streptomyces / genetics
  • Teicoplanin / biosynthesis*
  • Teicoplanin / metabolism
  • Teicoplanin / therapeutic use

Substances

  • Anti-Bacterial Agents
  • Teicoplanin
  • Glucuronidase