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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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