Cloning and characterization of the genes encoding enzymes for the protocatechuate meta-degradation pathway of Pseudomonas ochraceae NGJ1

Biosci Biotechnol Biochem. 2004 Jul;68(7):1434-41. doi: 10.1271/bbb.68.1434.

Abstract

The 2-pyrone-4,6-dicarboxylate lactonase gene (proL), the protocatechuate 4,5-dioxygenase alpha and beta subunits genes (proOa and proOb), and the 4-carboxy-2-hydroxymuconate-6-semialdehyde dehydrogenase gene (proD) were cloned from the chromosomal DNA of Pseudomonas ochraceae NGJ1. These genes were in the order proLOaObD on the DNA, and a possible transcription terminator sequence followed. The proL and proD genes were over-expressed in Escherichia coli, and their gene products were purified for identification, while the expression of proOaOb was at a lower level. The protocatechuate meta-degradation operon was reconstituted with the recombinant plasmids and expressed successfully in E. coli.

MeSH terms

  • Aldehyde Oxidoreductases / genetics*
  • Aldehyde Oxidoreductases / metabolism
  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Dioxygenases / genetics*
  • Dioxygenases / metabolism
  • Escherichia coli / genetics
  • Hydroxybenzoates / metabolism*
  • Molecular Sequence Data
  • Molecular Weight
  • Mutagenesis, Site-Directed
  • Open Reading Frames
  • Operon
  • Pseudomonas / enzymology*
  • Pseudomonas / genetics*
  • Pseudomonas / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Sequence Analysis, DNA
  • Sequence Analysis, Protein

Substances

  • DNA, Bacterial
  • Hydroxybenzoates
  • Recombinant Proteins
  • protocatechuic acid
  • 2-hydroxy-4-carboxymucconate-6-semialdehyde dehydrogenase
  • Dioxygenases
  • protocatechuate 4,5-dioxygenase
  • Aldehyde Oxidoreductases

Associated data

  • GENBANK/AB127969