Characterization of Two Polyketide Synthase Genes in Exophiala Lecanii-Corni, a Melanized Fungus With Bioremediation Potential

Bioorg Chem. 2004 Apr;32(2):92-108. doi: 10.1016/j.bioorg.2003.10.001.

Abstract

Exophiala lecanii-corni has significant bioremediation potential because it can degrade a wide range of volatile organic compounds. In order to identify sites for the insertion of genes that might enhance this potential, a genetic analysis of E. lecanii-corni was undertaken. Two polyketide synthase genes, ElPKS1 and ElPKS2, have now been discovered by a PCR-based strategy. ElPKS1 was isolated by a marker rescue technique. The nucleotide sequence of ElPKS1 consists of a 6576-bp open reading frame encoding a protein with 2192 amino acids, which was interrupted by a 60-bp intron near the 5' end and a 54-bp intron near the 3' end. Sequence analysis, results from disruption experiments, and physiological tests showed that ElPKS1 encoded a polyketide synthase required for melanin biosynthesis. Since ElPKS1 is non-essential, it is a desirable bioengineering target site for the insertion of native and foreign genes. The successful expression of these genes could enhance the bioremediation capability of the organism. ElPKS2 was cloned by colony hybridization screening of a partial genomic library with an ElPKS2 PCR product. ElPKS2 had a 6465-bp open reading frame that encoded 2155 amino acids and had introns of 56, 67, 54, and 71 bp. Although sequence analysis of the derived protein of ElPKS2 confirmed the polyketide synthase nature of its protein product, the function of that product remains unclear.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Biodegradation, Environmental
  • Cloning, Molecular
  • Exophiala / enzymology*
  • Exophiala / genetics*
  • Exophiala / growth & development
  • Genes, Fungal / genetics*
  • Introns / genetics
  • Molecular Sequence Data
  • Multienzyme Complexes / chemistry*
  • Multienzyme Complexes / genetics*
  • Multienzyme Complexes / metabolism
  • Mutation / genetics
  • Phenotype
  • Phylogeny
  • Polymerase Chain Reaction
  • Sequence Alignment

Substances

  • Multienzyme Complexes