Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Yamadazyma tenuis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | a two-enzyme system composed of meso-2,3-butanediol dehydrogenase (BDH) and xylose reductase is constructed to co-produce acetoin and xylitol with NAD+ regeneration. Four BDHs from four candidate organisms (Bacillus subtilis, Corynebacterium glutamicum, Parageobacillus thermoglucosidans, and Pyrococcus furiosus), as well as xylose reductase from Candida tenuis are purified and analyzed The best BDH is then selected according to titers and chiral purities of acetoin. After optimization of reaction conditions, and the ratios of meso-2,3-butanediol to xylose and BDH to xylose reductase, 28.5 g/l D-(-)-acetoin with an optical purity of 95.2% is produced in 6 h. The yield and productivity of acetoin is 0.97 g/g and 4.75 g/l/h. The titer of co-product xylitol is 40.29 g/l, and the yield and productivity of xylitol reaches 0.98 g/g and 6.72 g/l/h. Method development, evaluation, and optimization for production of optically pure D-(-)-acetoin, overview. Enzyme CT-XR acts most effectively with BDH from Corynebacterium glutamicum (CG-BDH) | Yamadazyma tenuis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | no substrate inhibition by D-xylose | Yamadazyma tenuis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
NADH | pH 6.5, 42°C, recombinant His-tagged enzyme | Yamadazyma tenuis | |
29.8 | - |
D-xylose | pH 6.5, 42°C, recombinant His-tagged enzyme | Yamadazyma tenuis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-xylose + NADH + H+ | Yamadazyma tenuis | - |
xylitol + NAD+ | - |
r | |
D-xylose + NADPH + H+ | Yamadazyma tenuis | - |
xylitol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Yamadazyma tenuis | O74237 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration | Yamadazyma tenuis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
7.12 | - |
purified recombinant His-tagged enzyme, pH 6.5, 42°C | Yamadazyma tenuis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-xylose + NADH + H+ | - |
Yamadazyma tenuis | xylitol + NAD+ | - |
r | |
D-xylose + NADPH + H+ | - |
Yamadazyma tenuis | xylitol + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 39000, about, recombinant His-tagged enzyme, SDS-PAGE, x * 36000, about, sequence calculation | Yamadazyma tenuis |
Synonyms | Comment | Organism |
---|---|---|
CT-XR | - |
Yamadazyma tenuis |
NAD(P)H-dependent D-xylose reductase | UniProt | Yamadazyma tenuis |
NADH-dependent XR | - |
Yamadazyma tenuis |
XYL1 | - |
Yamadazyma tenuis |
xylose reductase | - |
Yamadazyma tenuis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
42 | - |
- |
Yamadazyma tenuis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
27 | 57 | activity range | Yamadazyma tenuis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | 42 | purified recombinant His-tagged enzyme CT-XR, unstable at 42°C, complete inactivation after 6 h, CT-XR is more stable at 37°C maintaining 37.4% of initial activity after 6 h | Yamadazyma tenuis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11.42 | - |
NADH | pH 6.5, 42°C, recombinant His-tagged enzyme | Yamadazyma tenuis | |
11.42 | - |
D-xylose | pH 6.5, 42°C, recombinant His-tagged enzyme | Yamadazyma tenuis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
- |
Yamadazyma tenuis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5.5 | 9 | activity range | Yamadazyma tenuis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | most XRs are NADPH-dependent rather than NADH-dependent. CT-XR from Candida tenuis shows a similar preference for both NADH and NADPH | Yamadazyma tenuis | |
NAD+ | - |
Yamadazyma tenuis | |
NADH | - |
Yamadazyma tenuis | |
NADP+ | - |
Yamadazyma tenuis | |
NADPH | - |
Yamadazyma tenuis |
General Information | Comment | Organism |
---|---|---|
physiological function | most XRs are NADPH-dependent rather than NADH-dependent. CT-XR from Candida tenuis shows a similar preference for both NADH and NADPH, and has been reported to efficiently produce xylitol in vitro | Yamadazyma tenuis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.383 | - |
D-xylose | pH 6.5, 42°C, recombinant His-tagged enzyme | Yamadazyma tenuis | |
285.5 | - |
NADH | pH 6.5, 42°C, recombinant His-tagged enzyme | Yamadazyma tenuis |