Metabolic diversity in myxobacteria: identification of the myxalamid and the stigmatellin biosynthetic gene cluster of Stigmatella aurantiaca Sg a15 and a combined polyketide-(poly)peptide gene cluster from the epothilone producing strain Sorangium cellulosum So ce90

Biochim Biophys Acta. 1999 May 14;1445(2):185-95. doi: 10.1016/s0167-4781(99)00041-x.

Abstract

Myxobacterial strains producing polyketides (PKs) assumed to be biosynthesized by a type I polyketide synthase (PKS) were analysed. Myxobacteria also produce a variety of polypeptides (PP) and PKs with incorporated amino acids ('mixed PK-PP'). In order to be able to identify the biosynthetic gene clusters for these metabolites a PCR based approach has been developed to clone ketosynthase (KS) domains of PKS genes from these organisms. Conserved regions of peptide synthetases of the non-ribosomal type (NRPS) were also amplified via PCR. KS fragments from Stigmatella aurantiaca Sg a15 were used for chromosomal gene inactivation experiments resulting in a series of mutants including such that were unable to produce stigmatellins and myxalamids. A NRPS fragment and PKS fragments from Sorangium cellulosum So ce90 were used to identify cosmids hybridizing with both types of probes from a genomic library. Both a NRPS and a PKS fragment were cloned and sequenced from a relatively short restriction fragment of one of these cosmids. The method described here should be very useful to clone and identify PKS, NRPS and mixed PKS-NRPS from myxobacteria in general and thereby open opportunities to use the biochemical diversity of these bacteria for genetic engineering and combinatorial biosynthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cloning, Molecular
  • DNA, Bacterial / biosynthesis
  • Molecular Sequence Data
  • Multienzyme Complexes / biosynthesis
  • Multienzyme Complexes / genetics*
  • Multigene Family
  • Myxococcales / genetics*
  • Myxococcales / metabolism
  • Peptide Synthases / biosynthesis
  • Peptide Synthases / genetics
  • Plasmids
  • Polyenes / metabolism
  • Sequence Alignment

Substances

  • DNA, Bacterial
  • Multienzyme Complexes
  • Polyenes
  • stigmatellin
  • Peptide Synthases
  • non-ribosomal peptide synthase