Metabolic engineering of Pseudomonas putida for methylmalonyl-CoA biosynthesis to enable complex heterologous secondary metabolite formation

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Frank Gross - , Saarland University, Gesellschaft für Biotechnologische Forschung mbH (GBF), Gene Bridges GmbH (Author)
  • Michael W Ring - , Saarland University (Author)
  • Olena Perlova - , Saarland University, Gesellschaft für Biotechnologische Forschung mbH (GBF) (Author)
  • Jun Fu - , Chair of Applied Genomics, Gene Bridges GmbH (Author)
  • Susan Schneider - , TUD Dresden University of Technology (Author)
  • Klaus Gerth - , Gesellschaft für Biotechnologische Forschung mbH (GBF) (Author)
  • Silvia Kuhlmann - , Saarland University (Author)
  • A Francis Stewart - , Chair of Applied Genomics (Author)
  • Youming Zhang - , Gene Bridges GmbH (Author)
  • Rolf Müller - , Saarland University (Author)

Abstract

An operon consisting of three open reading frames, annotated in silico as methylmalonyl-CoA (mm-CoA) epimerase, mm-CoA mutase (MCM), and meaB, was identified in the sequencing project of the myxobacterium Sorangium cellulosum So ce56. This putative MCM pathway operon was subcloned from a bacterial artificial chromosome by Red/ET recombineering onto a minimal replicon derived from p15A. This plasmid was modified for integration and heterologous expression in Pseudomonas putida to enable the production of complex secondary metabolites requiring mm-CoA as precursor. Methylmalonate was identified in the recombinant P. putida strain by an analysis method based on gas chromatography/mass spectrometry. The engineered strain is able to synthesize polyketides requiring mm-CoA as an extender unit, which was demonstrated by the production of myxothiazol after integration of the biosynthetic gene cluster into the chromosome, followed by induction of expression.

Details

Original languageEnglish
Pages (from-to)1253-64
Number of pages12
JournalChemistry and Biology
Volume13
Issue number12
Publication statusPublished - Dec 2006
Peer-reviewedYes

External IDs

PubMed 17185221
Scopus 33845496065
ORCID /0000-0002-4754-1707/work/142248089

Keywords

Keywords

  • Acyl Coenzyme A/biosynthesis, Amino Acid Sequence, Gene Expression Regulation, Gene Transfer Techniques, Genetic Engineering/methods, Methacrylates/metabolism, Molecular Sequence Data, Myxococcales/genetics, Operon/genetics, Pseudomonas putida/genetics, Sequence Alignment, Thiazoles/metabolism