EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.1.B60 | expressed in Escherichia coli BL21(DE3)-CodonPlus RIL cells | Gluconobacter oxydans |
1.1.99.21 | gene sldh, subcloning in Escherichia coli strain DH5alpha, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)-CodonPlus RIL | Gluconobacter oxydans |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.99.21 | K294Q | site-directed mutagenesis, inactive mutant | Gluconobacter oxydans |
1.1.99.21 | additional information | stability of GoSLDH significantly improves up to 13.6fold after cross-linking of immobilized enzyme on silica nanoparticles and retains 62.8% residual activity after 10 cycles of reuse. Covalent immobilization of GoSLDH onto SiO2 nanoparticles: the amino groups of amino acids such as lysine present on the surface of GoSLDH react with the glutaraldehyde activates SiO2 nanoparticles to form covalent bonds during immobilization at pH 7. The IY and IE of GoSLDH immobilized on different silica nanoparticles are in the ranges of 40.4-71.2% and 53.5-76.7%, respectively | Gluconobacter oxydans |
EC Number | General Stability | Organism |
---|---|---|
1.1.1.B60 | stability of the enzyme significantly improves (up to 13.6fold) after cross-linking of immobilized enzyme on silica nanoparticles and retains 62.8% residual activity after 10 cycles of reuse | Gluconobacter oxydans |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.B60 | D-sorbitol | inhibitory at more than 10% (w/v) | Gluconobacter oxydans | |
1.1.99.21 | Ba2+ | - |
Gluconobacter oxydans | |
1.1.99.21 | Ca2+ | - |
Gluconobacter oxydans | |
1.1.99.21 | Cu2+ | - |
Gluconobacter oxydans | |
1.1.99.21 | K+ | - |
Gluconobacter oxydans | |
1.1.99.21 | additional information | EDTA treatment does not affect the initial enzyme activity | Gluconobacter oxydans | |
1.1.99.21 | Zn2+ | - |
Gluconobacter oxydans |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.B60 | 38.9 | - |
D-sorbitol | at pH 10.0 and 25°C | Gluconobacter oxydans | |
1.1.99.21 | 0.0882 | - |
NADP+ | pH 10.0, 25°C | Gluconobacter oxydans | |
1.1.99.21 | 38.9 | - |
D-sorbitol | pH 10.0, 25°C | Gluconobacter oxydans |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.B60 | Ba2+ | activates | Gluconobacter oxydans | |
1.1.1.B60 | Ca2+ | activates | Gluconobacter oxydans | |
1.1.1.B60 | Cu2+ | activates | Gluconobacter oxydans | |
1.1.1.B60 | K+ | activates | Gluconobacter oxydans | |
1.1.1.B60 | additional information | Mg2+, Co2+, and Mn2+ ions show no stimulatory influence on enzyme activity | Gluconobacter oxydans | |
1.1.1.B60 | Zn2+ | activates | Gluconobacter oxydans | |
1.1.99.21 | additional information | Mg2+, Co2+, and Mn2+ ions show no stimulatory influence on GoSLDH activity | Gluconobacter oxydans |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.1.1.B60 | 54000 | - |
2 * 54000, SDS-PAGE | Gluconobacter oxydans |
1.1.99.21 | 53640 | - |
- |
Gluconobacter oxydans |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.99.21 | D-sorbitol + NADP+ | Gluconobacter oxydans | - |
L-sorbose + NADPH | - |
r | |
1.1.99.21 | D-sorbitol + NADP+ | Gluconobacter oxydans G624 | - |
L-sorbose + NADPH | - |
r | |
1.1.99.21 | additional information | Gluconobacter oxydans | this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate | ? | - |
? | |
1.1.99.21 | additional information | Gluconobacter oxydans G624 | this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.1.B60 | Gluconobacter oxydans | - |
- |
- |
1.1.1.B60 | Gluconobacter oxydans G624 | - |
- |
- |
1.1.99.21 | Gluconobacter oxydans | - |
- |
- |
1.1.99.21 | Gluconobacter oxydans G624 | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.1.1.B60 | Ni-NTA column chromatography | Gluconobacter oxydans |
1.1.99.21 | recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)-CodonPlus RIL | Gluconobacter oxydans |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
1.1.99.21 | additional information | - |
Gluconobacter oxydans G624 shows SLDH activity of 43.2 U/mL and a 20% conversion yield from D-sorbitol to L-sorbose at 18 h of fermentation | Gluconobacter oxydans |
1.1.99.21 | 3570 | - |
purified recombinant enzyme, with D-sorbitol using NADP+ as a coenzyme, pH 10.0, 25°C | Gluconobacter oxydans |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.B60 | D-arabinitol + NADP+ | - |
Gluconobacter oxydans | ? | - |
? | |
1.1.1.B60 | D-sorbitol + NADP+ | best substrate | Gluconobacter oxydans | L-sorbose + NADPH + H+ | - |
? | |
1.1.1.B60 | D-sorbitol + NADP+ | best substrate | Gluconobacter oxydans G624 | L-sorbose + NADPH + H+ | - |
? | |
1.1.1.B60 | mannitol + NADP+ | - |
Gluconobacter oxydans | ? | - |
? | |
1.1.1.B60 | additional information | no activity is detected with L-arabinitol, xylitol, ribitol, myo-inositol, and glycerol | Gluconobacter oxydans | ? | - |
? | |
1.1.1.B60 | additional information | no activity is detected with L-arabinitol, xylitol, ribitol, myo-inositol, and glycerol | Gluconobacter oxydans G624 | ? | - |
? | |
1.1.99.21 | D-sorbitol + NADP+ | - |
Gluconobacter oxydans | L-sorbose + NADPH | - |
r | |
1.1.99.21 | D-sorbitol + NADP+ | - |
Gluconobacter oxydans G624 | L-sorbose + NADPH | - |
r | |
1.1.99.21 | additional information | this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate | Gluconobacter oxydans | ? | - |
? | |
1.1.99.21 | additional information | GoSLDH is highly specific towards D-sorbitol, mannitol, and D-arabinitol (Table S2). No activity is detected with L-arabinitol, xylitol, ribitol, myo-inositol, and glycerol | Gluconobacter oxydans | ? | - |
? | |
1.1.99.21 | additional information | this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate | Gluconobacter oxydans G624 | ? | - |
? | |
1.1.99.21 | additional information | GoSLDH is highly specific towards D-sorbitol, mannitol, and D-arabinitol (Table S2). No activity is detected with L-arabinitol, xylitol, ribitol, myo-inositol, and glycerol | Gluconobacter oxydans G624 | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.1.1.B60 | homodimer | 2 * 54000, SDS-PAGE | Gluconobacter oxydans |
1.1.99.21 | dimer | - |
Gluconobacter oxydans |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.1.B60 | D-sorbitol dehydrogenase (NADP+) | - |
Gluconobacter oxydans |
1.1.1.B60 | D-sorbitol:NADP+ oxidoreductase (L-sorbose-forming) | - |
Gluconobacter oxydans |
1.1.1.B60 | SLDH | - |
Gluconobacter oxydans |
1.1.99.21 | GoSLDH | - |
Gluconobacter oxydans |
1.1.99.21 | SLDH | - |
Gluconobacter oxydans |
1.1.99.21 | sorbitol dehydrogenase | - |
Gluconobacter oxydans |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.B60 | 70 | - |
- |
Gluconobacter oxydans |
1.1.99.21 | 70 | - |
oxidation reaction | Gluconobacter oxydans |
EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.B60 | 25 | 75 | more than 40% activity between 25 and 75°C | Gluconobacter oxydans |
1.1.99.21 | 20 | 80 | activity range, profile overview | Gluconobacter oxydans |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.B60 | 3820 | - |
D-sorbitol | at pH 10.0 and 25°C | Gluconobacter oxydans | |
1.1.99.21 | 3820 | - |
D-sorbitol | pH 10.0, 25°C | Gluconobacter oxydans |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.1.B60 | 10 | - |
- |
Gluconobacter oxydans |
1.1.99.21 | 10 | - |
oxidation reaction | Gluconobacter oxydans |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
1.1.99.21 | 7.5 | 11 | activity range, profile overview | Gluconobacter oxydans |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.B60 | NADP+ | the enzyme exhibits high preference for NADP+ (vs. 2.5% relative activity with NAD+) | Gluconobacter oxydans | |
1.1.99.21 | additional information | enzyme GoSLDH exhibits a preference for NADP+ over NAD+ as a coenzyme and is exclusively an NADP+-dependent enzyme showing only 2.5% relative activity with NAD+ | Gluconobacter oxydans | |
1.1.99.21 | NADP+ | - |
Gluconobacter oxydans | |
1.1.99.21 | NADPH | - |
Gluconobacter oxydans |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.1.99.21 | evolution | GoSLDH sequencing, structure analyses, and biochemical studies, suggest that it belongs to the NADP+-dependent polyol specific long-chain sorbitol dehydrogenase family. GoSLDH possesses the active site residues of Asp190, Val228, Lys294, and Asp299 and the conserved catalytic motif (KXXXXHXXH) for polyol-specific long-chain dehydrogenase | Gluconobacter oxydans |
1.1.99.21 | additional information | structure comparisons and molecular docking, overview | Gluconobacter oxydans |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.B60 | 98.1 | - |
D-sorbitol | at pH 10.0 and 25°C | Gluconobacter oxydans | |
1.1.99.21 | 98.1 | - |
D-sorbitol | pH 10.0, 25°C | Gluconobacter oxydans |