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J Bacteriol, July 1998, p. 3697-3703, Vol. 180, No. 14
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Molecular Cloning and Nucleotide Sequence of the
Superoxide Dismutase Gene and Characterization of Its Product from
Bacillus subtilis
Takashi
Inaoka,1
Yoshinobu
Matsumura,1,2,* and
Tetsuaki
Tsuchido1,2
Department of Biotechnology, Faculty of
Engineering,1 and
High-Technology
Research Center,2 Kansai University, Suita,
Osaka 564, Japan
Received 3 November 1997/Accepted 18 May 1998
Bacillus subtilis was found to possess one detectable
superoxide dismutase (Sod) in both vegetative cells and spores. The Sod
activity in vegetative cells was maximal at stationary phase. Manganese
was necessary to sustain Sod activity at stationary phase, but
paraquat, a superoxide generator, did not induce the expression of Sod.
The specific activity of purified Sod was approximately 2,600 U/mg of
protein, and the enzyme was a homodimer protein with a molecular mass
of approximately 25,000 per monomer. The gene encoding Sod, designated
sodA, was cloned by the combination of several PCR methods
and the Southern hybridization method. DNA sequence analysis revealed
the presence of one open reading frame consisting of 606 bp. Several
putative promoter sites were located in the upstream region of
sodA. The deduced amino acid sequence showed high homology
with other bacterial manganese Sods. Conserved regions in bacterial
manganese Sod could also be seen. The phenotype of double mutant
Escherichia coli sodA sodB, which could not grow in minimal
medium without supplemental amino acids, was complemented by the
expression of B. subtilis sodA.
*
Corresponding author. Mailing address: Department of
Biotechnology, Faculty of Engineering, Kansai University, 3-3-35 Yamate-cho, Suita, Osaka 564, Japan. Phone: 81-6-368-0934. Fax:
81-6-388-8609. E-mail: ymatsu{at}ipcku.kansai-u.ac.jp.
J Bacteriol, July 1998, p. 3697-3703, Vol. 180, No. 14
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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