Kim, Tae-Su Patel, Sanjay K. S. Selvaraj, Chandrabose Jung, Woo-Suk Pan, Cheol-Ho Kang, Yun Chan Lee, Jung-Kul 2024-01-20T03:31:22Z 2024-01-20T03:31:22Z 2021-08-31 2016-09-16 2045-2322 https://pubs.kist.re.kr/handle/201004/123670 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 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. English NATURE PUBLISHING GROUP L-SORBOSE MANNITOL 2-DEHYDROGENASE PSEUDOMONAS-FLUORESCENS ENZYME IMMOBILIZATION CATALYTIC MECHANISM MOLECULAR-CLONING DOCKING INHIBITORS EXPRESSION STRATEGIES A highly efficient sorbitol dehydrogenase from Gluconobacter oxydans G624 and improvement of its stability through immobilization Article 10.1038/srep33438 1 SCIENTIFIC REPORTS, v.6 SCIENTIFIC REPORTS 6 scie scopus 000383646300001 2-s2.0-84988422978 Multidisciplinary Sciences Science & Technology - Other Topics Article L-SORBOSE MANNITOL 2-DEHYDROGENASE PSEUDOMONAS-FLUORESCENS ENZYME IMMOBILIZATION CATALYTIC MECHANISM MOLECULAR-CLONING DOCKING INHIBITORS EXPRESSION STRATEGIES sorbitol dehydrogenase Gluconobacter oxydans G624 L-sorbose-producing enzyme substrate specificity immobilization