1887

Abstract

The two genes for catechol 2,3-oxygenase, encoded by TOL plasmid pWW53, carry a common I restriction site within the reading frame. Each gene was cut at the I site and the 5′ end of each gene spliced to the 3′ end of the other to form hybrid genes, from both of which catalytically active catechol 2,3-oxygenase activities were expressed. The kinetic parameters were determined for the gene products of both the hybrid and the wild-type genes with catechol, 3-methylcatechol and 4-methylcatechol as substrates. Comparison of the results suggested firstly, that the C-terminal regions of the enzymes determined both the binding and the catalytic specificity, and, secondly, that the N-terminal region of one of the enzymic gene products contained a secondary binding site which caused inhibition by excess substrate for methylcatechol substrates but not for catechol. One of the wild-type enzymes appeared to have an intrinsically higher activity for all three substrates than the other. This higher activity depended on the presence of both its C- and N-terminal regions, and in both hybrid enzymes, which contained only one of these regions, activity was significantly reduced.

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/content/journal/micro/10.1099/00221287-136-8-1583
1990-08-01
2021-07-30
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