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ID 111972
Author
Ohshida, Tatsuya Kagawa University
Satomura, Takenori University of Fukui
Sakuraba, Haruhiko Kagawa University
Keywords
halophile
Haloarcula japonica
2-deoxy-D-ribose-5-phosphate aldolase
archaea
organic solvent
aldehyde
Content Type
Journal Article
Description
2-Deoxy-D-ribose-5-phosphate aldolase (DERA) catalyzes the aldol reaction between two aldehydes and is thought to be a potential biocatalyst for the production of a variety of stereo-specific materials. A gene encoding DERA from the extreme halophilic archaeon, Haloarcula japonica, was overexpressed in Escherichia coli. The gene product was successfully purified, using procedures based on the protein’s halophilicity, and characterized. The expressed enzyme was stable in a buffer containing 2 M NaCl and exhibited high thermostability, retaining more than 90% of its activity after heating at 70℃ for 10 min. The enzyme was also tolerant to high concentrations of organic solvents, such as acetonitrile and dimethylsulfoxide. Moreover, H. japonica DERA was highly resistant to a high concentration of acetaldehyde and retained about 35% of its initial activity after 5-hours’ exposure to 300 mM acetaldehyde at 25℃, the conditions under which E. coli DERA is completely inactivated. The enzyme exhibited much higher activity at 25℃ than the previously characterized hyperthermophilic DERAs (Sakuraba et al., 2007). Our results suggest that the extremely halophilic DERA has high potential to serve as a biocatalyst in organic syntheses. This is the first description of the biochemical characterization of a halophilic DERA.
Journal Title
Protein Expression and Purification
ISSN
10465928
NCID
AA10814149
AA11542088
Publisher
Elsevier
Volume
126
Start Page
62
End Page
68
Published Date
2016-05-20
Rights
© 2016. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
EDB ID
DOI (Published Version)
URL ( Publisher's Version )
FullText File
language
eng
TextVersion
Author
departments
Bioscience and Bioindustry