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Applied and Environmental Microbiology, October 2002, p. 4841-4846, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4841-4846.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of a Hydratase and a Class II Aldolase Involved in Biodegradation of the Organic Solvent Tetralin

M. J. Hernáez,1,{dagger} B. Floriano,1,{ddagger} J. J. Ríos,2 and E. Santero1*

Departamento de Genética, Facultad de Biología, Universidad de Sevilla,1 Instituto de la Grasa, CSIC, Seville, Spain2

Received 15 April 2002/ Accepted 15 July 2002

Two new genes whose products are involved in biodegradation of the organic solvent tetralin were identified. These genes, designated thnE and thnF, are located downstream of the previously identified thnD gene and code for a hydratase and an aldolase, respectively. A sequence comparison of enzymes similar to ThnE showed the significant similarity of hydratases involved in biodegradation pathways to 4-oxalocrotonate decarboxylases and established four separate groups of related enzymes. Consistent with the sequence information, characterization of the reaction catalyzed by ThnE showed that it hydrated a 10-carbon dicarboxylic acid. The only reaction product detected was the enol tautomer, 2,4-dihydroxydec-2-ene-1,10-dioic acid. The aldolase ThnF showed significant similarity to aldolases involved in different catabolic pathways whose substrates are dihydroxylated dicarboxylic acids and which yield pyruvate and a semialdehyde. The reaction products of the aldol cleavage reaction catalyzed by ThnF were identified as pyruvate and the seven-carbon acid pimelic semialdehyde. ThnF and similar aldolases showed conservation of the active site residues identified by the crystal structure of 2-dehydro-3-deoxy-galactarate aldolase, a class II aldolase with a novel reaction mechanism, suggesting that these similar enzymes are class II aldolases. In contrast, ThnF did not show similarity to 4-hydroxy-2-oxovalerate aldolases of other biodegradation pathways, which are significantly larger and apparently are class I aldolases.


* Corresponding author. Present address: Laboratorio Andaluz de Biología, Universidad Pablo de Olavide, ctra. Utrera Km 1, 41013 Seville, Spain. Phone: 34-95-4349386. Fax: 34-95-4349376. E-mail: esansan{at}dex.upo.es.

{dagger} Present address: Instituto Valenciano de Infertilidad, Seville, Spain.

{ddagger} Present address: Laboratorio Andaluz de Biología, Universidad Pablo de Olavide, Seville, Spain.


Applied and Environmental Microbiology, October 2002, p. 4841-4846, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4841-4846.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.




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