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