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Applied and Environmental Microbiology, April 2000, p. 1499-1508, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of the Protocatechuic Acid Catabolic Gene Cluster from Streptomyces sp. Strain 2065

Sakura G. Iwagami, Keqian Yang, and Julian Davies*

Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada

Received 31 August 1999/Accepted 1 February 2000

Protocatechuate 3,4-dioxygenase (EC 1.13.11.3) catalyzes the ring cleavage step in the catabolism of aromatic compounds through the protocatechuate branch of the beta -ketoadipate pathway. A protocatechuate 3,4-dioxygenase was purified from Streptomyces sp. strain 2065 grown in p-hydroxybenzoate, and the N-terminal sequences of the beta - and alpha -subunits were obtained. PCR amplification was used for the cloning of the corresponding genes, and DNA sequencing of the flanking regions showed that the pcaGH genes belonged to a 6.5-kb protocatechuate catabolic gene cluster; at least seven genes in the order pcaIJFHGBL appear to be transcribed unidirectionally. Analysis of the cluster revealed the presence of a pcaL homologue which encodes a fused gamma -carboxymuconolactone decarboxylase/beta -ketoadipate enol-lactone hydrolase previously identified in the pca gene cluster from Rhodococcus opacus 1CP. The pcaIJ genes encoded proteins with a striking similarity to succinyl-coenzyme A (CoA):3-oxoacid CoA transferases of eukaryotes and contained an indel which is strikingly similar between high-G+C gram-positive bacteria and eukaryotes.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, 300-6041 University Blvd., Vancouver, British Columbia, V6T 1Z1, Canada. Phone: 1 604 221-8896. Fax: 1 604 822 6041. E-mail: jed{at}interchange.ubc.ca.

Applied and Environmental Microbiology, April 2000, p. 1499-1508, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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