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Journal of Bacteriology, October 2000, p. 5448-5453, Vol. 182, No. 19
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification of a Serine Hydrolase Which Cleaves the Alicyclic Ring of Tetralin

M. J. Hernáez,1 E. Andújar,1 J. L. Ríos,2 S. R. Kaschabek,3 W. Reineke,3 and E. Santero1,*

Departamento de Genética, Facultad de Biología, Universidad de Sevilla,1 and Instituto de la Grasa, CSIC,2 Seville, Spain, and Chemische Mikrobiologie, Bergische Universität-Gesamthochschule Wuppertal, Wuppertal, Germany3

Received 13 April 2000/Accepted 28 June 2000

A gene designated thnD, which is required for biodegradation of the organic solvent tetralin by Sphingomonas macrogoltabidus strain TFA, has been identified. Sequence comparison analysis indicated that thnD codes for a carbon-carbon bond serine hydrolase showing highest similarity to hydrolases involved in biodegradation of biphenyl. An insertion mutant defective in ThnD accumulates the ring fission product which results from the extradiol cleavage of the aromatic ring of dihydroxytetralin. The gene product has been purified and characterized. ThnD is an octameric thermostable enzyme with an optimum reaction temperature at 65°C. ThnD efficiently hydrolyzes the ring fission intermediate of the tetralin pathway and also 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid, the ring fission product of the biphenyl meta-cleavage pathway. However, it is not active towards the equivalent intermediates of meta-cleavage pathways of monoaromatic compounds which have small substituents in C-6. When ThnD hydrolyzes the intermediate in the tetralin pathway, it cleaves a C-C bond comprised within the alicyclic ring of tetralin instead of cleaving a linear C-C bond, as all other known hydrolases of meta-cleavage pathways do. The significance of this activity of ThnD for the requirement of other activities to mineralize tetralin is discussed.

* Corresponding author. Mailing address: Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Ap. 1095, 41080 Seville, Spain. Phone: 34-95-4557106. Fax: 34-95-4557104. E-mail: esantero{at}cica.es.

Journal of Bacteriology, October 2000, p. 5448-5453, Vol. 182, No. 19
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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