5ITG

Crystal structure of D-sorbitol dehydrogenase in substrate-free form

5ITG の概要

• エントリーDOI: 10.2210/pdb5itg/pdb
• 分子名称: Sorbitol dehydrogenase (2 entities in total)
• 機能のキーワード: dehydrogenase, rossmann-fold, l-sorbose, nadph cofactor, oxidoreductase
• 由来する生物種: Gluconobacter oxydans
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 108309.12

構造登録者

Jung, W.S.,Pan, C.H. (登録日: 2016-03-16, 公開日: 2017-03-08, 最終更新日: 2024-10-23)

主引用文献

Kim, T.S.,Patel, S.K.,Selvaraj, C.,Jung, W.S.,Pan, C.H.,Kang, Y.C.,Lee, J.K.
A highly efficient sorbitol dehydrogenase from Gluconobacter oxydans G624 and improvement of its stability through immobilization
Sci Rep, 6:33438-33438, 2016

PubMed Abstract: 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 Km and kcat 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.

PubMed: 27633501
DOI: 10.1038/srep33438

実験手法

X-RAY DIFFRACTION (1.95 Å)