A highly efficient sorbitol dehydrogenase from Gluconobacter oxydans G624 and improvement of its stability through immobilization

A sorbitol dehydrogenase (GoSLDH) from Gluconobacter oxydans G624 ( G. oxydans G624) was expressed in Escherichia coli BL21(DE3)-CodonPlus RIL. The complete 1455-bp codon-optimized gene was amplified, expressed, and thoroughly characterized for the first time. GoSLDH exhibited K m and k cat values o...

Full description

Saved in:
Bibliographic Details
Published in:Scientific reports 2016-09, Vol.6 (1), p.33438-33438, Article 33438
Main Authors: Kim, Tae-Su, Patel, Sanjay K. S., Selvaraj, Chandrabose, Jung, Woo-Suk, Pan, Cheol-Ho, Kang, Yun Chan, Lee, Jung-Kul
Format: Article
Language:English
Subjects:
631/45/603
631/61/338/318
Calorimetry
D-Sorbitol
Dehydrogenase
Dehydrogenases
E coli
Enzymes
Humanities and Social Sciences
Immobilization
multidisciplinary
NADP
Nanoparticles
Oxidation
Science
Silica
Sorbose dehydrogenase
Substrate specificity
Substrates
Titration
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A sorbitol dehydrogenase (GoSLDH) from Gluconobacter oxydans G624 ( G. oxydans G624) was expressed in Escherichia coli BL21(DE3)-CodonPlus RIL. The complete 1455-bp codon-optimized gene was amplified, expressed, and thoroughly characterized for the first time. GoSLDH exhibited K m and k cat values of 38.9 mM and 3820 s −1 toward L-sorbitol, respectively. The enzyme exhibited high preference for NADP + (vs. only 2.5% relative activity with NAD + ). GoSLDH sequencing, structure analyses, and biochemical studies, suggested that it belongs to the NADP + -dependent polyol-specific long-chain sorbitol dehydrogenase family. GoSLDH is the first fully characterized SLDH to date, and it is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry showed that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirmed a higher turnover rate. The high oxidation potential of GoSLDH for D-sorbitol was confirmed by cyclovoltametric analysis. Further, stability of GoSLDH significantly improved (up to 13.6-fold) after cross-linking of immobilized enzyme on silica nanoparticles and retained 62.8% residual activity after 10 cycles of reuse. Therefore, immobilized GoSLDH may be useful for L-sorbose production from D-sorbitol.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep33438