First characterization of extremely halophilic 2-deoxy-D-ribose-5-phosphate aldolase.

Abstract:

: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 °C 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-h' exposure to 300 mM acetaldehyde at 25 °C, the conditions under which E. coli DERA is completely inactivated. The enzyme exhibited much higher activity at 25 °C 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_name

Protein Expr Purif

authors

Ohshida T,Hayashi J,Satomura T,Kawakami R,Ohshima T,Sakuraba H

doi

10.1016/j.pep.2016.05.009

subject

Has Abstract

pub_date

2016-10-01 00:00:00

pages

62-68

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(16)30087-0

journal_volume

126

pub_type

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