TY - JOUR
T1 - An efficient ribitol-specific dehydrogenase from Enterobacter aerogenes
AU - Singh, Ranjitha
AU - Singh, Raushan
AU - Kim, In Won
AU - Sigdel, Sujan
AU - Kalia, Vipin C.
AU - Kang, Yun Chan
AU - Lee, Jung Kul
PY - 2015/5/1
Y1 - 2015/5/1
N2 - An NAD+-dependent ribitol dehydrogenase from Enterobacter aerogenes KCTC 2190 (EaRDH) was cloned and successfully expressed in Escherichia coli. The complete 729-bp gene was amplified, cloned, expressed, and subsequently purified in an active soluble form using nickel affinity chromatography. The enzyme had an optimal pH and temperature of 11.0 and 45°C, respectively. Among various polyols, EaRDH exhibited activity only toward ribitol, with Km, Vmax, and kcat/Km values of 10.3mM, 185Umg-1, and 30.9s-1mM-1, respectively. The enzyme showed strong preference for NAD+ and displayed no detectable activity with NADP+. Homology modeling and sequence analysis of EaRDH, along with its biochemical properties, confirmed that EaRDH belongs to the family of NAD+-dependent ribitol dehydrogenases, a member of short-chain dehydrogenase/reductase (SCOR) family. EaRDH showed the highest activity and unique substrate specificity among all known RDHs. Homology modeling and docking analysis shed light on the molecular basis of its unusually high activity and substrate specificity.
AB - An NAD+-dependent ribitol dehydrogenase from Enterobacter aerogenes KCTC 2190 (EaRDH) was cloned and successfully expressed in Escherichia coli. The complete 729-bp gene was amplified, cloned, expressed, and subsequently purified in an active soluble form using nickel affinity chromatography. The enzyme had an optimal pH and temperature of 11.0 and 45°C, respectively. Among various polyols, EaRDH exhibited activity only toward ribitol, with Km, Vmax, and kcat/Km values of 10.3mM, 185Umg-1, and 30.9s-1mM-1, respectively. The enzyme showed strong preference for NAD+ and displayed no detectable activity with NADP+. Homology modeling and sequence analysis of EaRDH, along with its biochemical properties, confirmed that EaRDH belongs to the family of NAD+-dependent ribitol dehydrogenases, a member of short-chain dehydrogenase/reductase (SCOR) family. EaRDH showed the highest activity and unique substrate specificity among all known RDHs. Homology modeling and docking analysis shed light on the molecular basis of its unusually high activity and substrate specificity.
KW - Enterobacter aerogenes
KW - Homology modeling
KW - Ribitol dehydrogenase
KW - Short-chain dehydrogenase/reductase
UR - http://www.scopus.com/inward/record.url?scp=84924869389&partnerID=8YFLogxK
U2 - 10.1016/j.enzmictec.2015.02.004
DO - 10.1016/j.enzmictec.2015.02.004
M3 - Journal article
C2 - 25837508
AN - SCOPUS:84924869389
SN - 0141-0229
VL - 72
SP - 56
EP - 64
JO - Enzyme and Microbial Technology
JF - Enzyme and Microbial Technology
ER -