Molecular cloning of groESL locus, and purification and characterization of chaperonins, GroEL and GroES, from Bacillus brevis.

Article abstract of DOI:10.1271/bbb.65.1379

The groESL locus of a protein-hypersecreting bacterium, Bacillus brevis, was cloned by PCR using primers designed based on the DNA sequence of a B. subtilis homolog. GroEL protein was purified to apparent homogeneity and its ATPase activity was characterized: it hydrolyzed ATP, CTP, and TTP in this order of reaction rate, and its specific activity for ATP was 0.1 micromole/min/mg protein. Purified GroEL forms a tetradecamer. GroEL was estimated to contain 22% alpha-helix, 24% beta-sheet, and 19% turn structures, by CD measurement. GroES protein was also highly purified to examine its chaperonin activity. GroEL protected from thermal inactivation of and showed refolding-promoting activity for malate dehydrogenase, strictly depending on the presence of ATP and GroES.

Full text of DOI:10.1271/bbb.65.1379

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Molecular Cloning of groESL Locus, and Purification
and Characterization of Chaperonins, GroEL and
GroES, from Bacillus brevis
a
a
a
b
Masao TOKUNAGA , Yoichi SHIRAISHI , Masatake ODACHI , Makoto MIZUKAMI , Hiroko
a
c
c
a
a
TOKUNAGA , John S. PHILO , Tsutomu ARAKAWA , Matsujiro ISHIBASHI , Ryoichi TANAKA
b
&
a
Hiroaki TAKAGI
Laboratory of Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima
University
b
Department of Research & Development, Higeta Shoyu Co.
c
Alliance Protein Laboratories
Published online: 22 May 2014.
To cite this article: Masao TOKUNAGA, Yoichi SHIRAISHI, Masatake ODACHI, Makoto MIZUKAMI, Hiroko TOKUNAGA,
John S. PHILO, Tsutomu ARAKAWA, Matsujiro ISHIBASHI, Ryoichi TANAKA & Hiroaki TAKAGI (2001) Molecular Cloning of
groESL Locus, and Purification and Characterization of Chaperonins, GroEL and GroES, from Bacillus brevis, Bioscience,
Biotechnology, and Biochemistry, 65:6, 1379-1387, DOI: 10.1271/bbb.65.1379
To link to this article: http://dx.doi.org/10.1271/bbb.65.1379
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