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

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Frank Gross - , Universität des Saarlandes, Gesellschaft für Biotechnologische Forschung mbH (GBF), Gene Bridges GmbH (Autor:in)
  • Michael W Ring - , Universität des Saarlandes (Autor:in)
  • Olena Perlova - , Universität des Saarlandes, Gesellschaft für Biotechnologische Forschung mbH (GBF) (Autor:in)
  • Jun Fu - , Professur für Biotechnologische Genomik, Gene Bridges GmbH (Autor:in)
  • Susan Schneider - , Technische Universität Dresden (Autor:in)
  • Klaus Gerth - , Gesellschaft für Biotechnologische Forschung mbH (GBF) (Autor:in)
  • Silvia Kuhlmann - , Universität des Saarlandes (Autor:in)
  • A Francis Stewart - , Professur für Biotechnologische Genomik (Autor:in)
  • Youming Zhang - , Gene Bridges GmbH (Autor:in)
  • Rolf Müller - , Universität des Saarlandes (Autor:in)

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

OriginalspracheEnglisch
Seiten (von - bis)1253-64
Seitenumfang12
FachzeitschriftChemistry and Biology
Jahrgang13
Ausgabenummer12
PublikationsstatusVeröffentlicht - Dez. 2006
Peer-Review-StatusJa

Externe IDs

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

Schlagworte

Schlagwörter

  • 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