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

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.

SnO2 hollow nanotubes: a novel and efficient support matrix for enzyme immobilization

“…Therefore, in the present study, unique SnO 2 hollow nanotubes were synthesized by electrospinning methods and used for the first time as supports for the immobilization of enzymes, including lipase, HRP, and GOx. Compared with other nanoparticles reported previously 17 , 20 , 36 – 40 , the synthesized SnO 2 hollow nanotubes were found to be superior supports for immobilizing a variety of enzymes owing to their large surface areas, additional surface porosities, and luminal openings. The improved immobilization yield and reusability of the SnO 2 nanotube-bound enzymes indicate that these hollow nanotubes are promising supports for enzyme immobilization.…”

Development of enzymatically-active bacterial cellulose membranes through stable immobilization of an engineered β-galactosidase

“…There is a wide range of enzyme immobilization techniques involving physical entrapment or chemical interactions (either covalent or non-covalent) between the enzyme and the support [9,[12][13][14][15][16]. These techniques have been applied to many enzymes, including -galactosidases [5,9,12,[17][18][19][20][21][22][23].…”

Canna edulis Leaf Extract-Mediated Preparation of Stabilized Silver Nanoparticles: Characterization, Antimicrobial Activity, and Toxicity Studies

“…Standard "green" science methods, which decrease the utilization or production of dangerous substances, are also being applied in the field of nanoscience, including the synthesis of nanoscale items, leading to the advancement of techniques for creation of nanomaterials [1][2][3][4]. A number of chemical and physical methods have been used to synthesize metal nanoparticles, including chemical reduction [5,6], electrochemical reduction [7][8][9], photochemical reduction [10], radiation [11,12], and heat evaporation [13,14].…”