Molecular cloning of the protocatechuate 4,5-dioxygenase genes of Pseudomonas paucimobilis

Y. Noda, S. Nishikawa, K. I. Shiozuka, H. Kadokura, H. Nakajima, K. Yoda, Y. Katayama, N. Morohoshi, T. Haraguchi, M. Yamasaki

Research output: Contribution to journalArticlepeer-review

106 Citations (Scopus)

Abstract

We determine the nucleotide sequence of a 1.9-kilobase fragment of Pseudomonas paucimobilis SYK6 chromosomal DNA that included genes encoding protocatechuate 4,5-dioxygenase, the enzyme responsible for the aromatic ring fission of protocatechuate. Two open reading frames of 417 and 906 base pairs were found that had no homology with previously reported sequences, including those encoding protocatechuate 3,4-dioxygenase. Since both open reading frames were indispensable for the enzyme activity, they should encode the subunits of protocatechuate 4,5-dioxygenase. We named these genes ligA and ligB. Protocatechuate 4,5-dioxygenase was efficiently expressed in Escherichia coli with the aid of the lac promoter, and the polypeptides of the ligA and ligB gene products were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and amino acid sequencing.

Original languageEnglish
Pages (from-to)2704-2709
Number of pages6
JournalJournal of bacteriology
Volume172
Issue number5
DOIs
Publication statusPublished - 1990

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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