Journal
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume 68, Issue 7, Pages 1434-1441Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1271/bbb.68.1434
Keywords
protocatechuate meta-degradation operon; protocatechuate 4,5-dioxygenase; 2-pyrone-4,6-dicarboxylate
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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.
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