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Applied and Environmental Microbiology, September 1999, p. 3774-3779, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Role of N
-Acetyldiaminobutyrate as an
Enzyme Stabilizer and an Intermediate in the Biosynthesis of
Hydroxyectoine
David
Cánovas,1,2,
Nuno
Borges,2
Carmen
Vargas,1
Antonio
Ventosa,1
Joaquín J.
Nieto,1,* and
Helena
Santos2
Departamento de Microbiología y
Parasitología, Facultad de Farmacia, Universidad de Sevilla,
41012 Seville, Spain,1 and Instituto de
Tecnologia Química e Biológica, Universidade Nova de
Lisboa, 2780 Oeiras, Portugal2
Received 7 April 1999/Accepted 8 June 1999
Strain CHR63 is a salt-sensitive mutant of the moderately
halophilic wild-type strain Halomonas elongata DSM 3043 that is affected in the ectoine synthase gene (ectC). This
strain accumulates large amounts of
N
-acetyldiaminobutyrate (NADA), the precursor of ectoine
(D. Cánovas, C. Vargas, F. Iglesias-Guerra, L. N. Csonka, D. Rhodes, A. Ventosa, and J. J. Nieto, J. Biol. Chem. 272:25794-25801, 1997). Hydroxyectoine, ectoine, and glucosylglycerate were also identified by nuclear magnetic resonance (NMR) as cytoplasmic organic solutes in this mutant. Accumulation of NADA, hydroxyectoine, and ectoine was osmoregulated, whereas the levels of glucosylglycerate decreased at higher salinities. The effect of the growth stage on the
accumulation of solutes was also investigated. NADA was purified from
strain CHR63 and was shown to protect the thermolabile enzyme rabbit
muscle lactate dehydrogenase against thermal inactivation. The
stabilizing effect of NADA was greater than the stabilizing effect of
ectoine or potassium diaminobutyrate. A 1H NMR analysis of
the solutes accumulated by the wild-type strain and mutants CHR62
(ectA::Tn1732) and CHR63
(ectC::Tn1732) indicated that
H. elongata can synthesize hydroxyectoine by two different pathways
directly from ectoine or via an alternative pathway that converts NADA into hydroxyectoine without the involvement of ectoine.
*
Corresponding author. Mailing address: Departamento de
Microbiología y Parasitología, Facultad de Farmacia,
Universidad de Sevilla, 41012 Seville, Spain. Phone: 34-954-556765. Fax: 34-954-628162. E-mail: jjnieto{at}cica.es.

Present address: Departamento de Biotecnología Microbiana,
Centro Nacional de Biotecnología, CSIC, Cantoblanco, 28049 Madrid,
Spain.
Applied and Environmental Microbiology, September 1999, p. 3774-3779, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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