Application | Comment | Organism |
---|---|---|
synthesis | Bacillus subtilis enzyme BS-BDH is a potential candidate for L-(+)-acetoin production | Bacillus subtilis |
Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Parageobacillus thermoglucosidasius |
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Pyrococcus furiosus |
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Bacillus subtilis |
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Corynebacterium glutamicum |
Protein Variants | Comment | Organism |
---|---|---|
additional information | a two-enzyme system composed of meso-2,3-butanediol dehydrogenase (BDH) and xylose reductase (CT-XR) from Candida tenuis 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) | Parageobacillus thermoglucosidasius |
additional information | a two-enzyme system composed of meso-2,3-butanediol dehydrogenase (BDH) and xylose reductase (CT-XR) from Candida tenuis 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) | Pyrococcus furiosus |
additional information | a two-enzyme system composed of meso-2,3-butanediol dehydrogenase (BDH) and xylose reductase (CT-XR) from Candida tenuis 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) | Bacillus subtilis |
additional information | a two-enzyme system composed of meso-2,3-butanediol dehydrogenase (BDH) and xylose reductase (CT-XR) from Candida tenuis 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) | Corynebacterium glutamicum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | no substrate inhibition by (2R,3S)-butane-2,3-diol | Bacillus subtilis | |
additional information | no substrate inhibition by (2R,3S)-butane-2,3-diol | Corynebacterium glutamicum | |
additional information | no substrate inhibition by (2R,3S)-butane-2,3-diol | Parageobacillus thermoglucosidasius | |
additional information | no substrate inhibition by (2R,3S)-butane-2,3-diol | Pyrococcus furiosus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
NAD+ | pH 9.0, 67°C, recombinant His-tagged enzyme | Parageobacillus thermoglucosidasius | |
0.12 | - |
NAD+ | pH 8.5, 47°C, recombinant His-tagged enzyme | Bacillus subtilis | |
0.2 | - |
NAD+ | pH 8.5, 52°C, recombinant His-tagged enzyme | Corynebacterium glutamicum | |
0.2 | - |
NAD+ | pH 9.0, 72°C, recombinant His-tagged enzyme | Pyrococcus furiosus | |
2.6 | - |
(2R,3S)-butane-2,3-diol | pH 8.5, 52°C, recombinant His-tagged enzyme | Corynebacterium glutamicum | |
27.7 | - |
(2R,3S)-butane-2,3-diol | pH 8.5, 47°C, recombinant His-tagged enzyme | Bacillus subtilis | |
27.7 | - |
(2R,3S)-butane-2,3-diol | pH 9.0, 67°C, recombinant His-tagged enzyme | Parageobacillus thermoglucosidasius | |
53.9 | - |
(2R,3S)-butane-2,3-diol | pH 9.0, 72°C, recombinant His-tagged enzyme | Pyrococcus furiosus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R,3S)-butane-2,3-diol + NAD+ | Bacillus subtilis | - |
(3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Corynebacterium glutamicum | - |
(3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Parageobacillus thermoglucosidasius | - |
(3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Pyrococcus furiosus | - |
(3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Corynebacterium glutamicum LMG 3730 | - |
(3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Corynebacterium glutamicum BCRC 11384 | - |
(3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Pyrococcus furiosus ATCC 43587 | - |
(3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Corynebacterium glutamicum ATCC 13032 | - |
(3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Pyrococcus furiosus Vc1 | - |
(3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Corynebacterium glutamicum JCM 1318 | - |
(3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Pyrococcus furiosus JCM 8422 | - |
(3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Corynebacterium glutamicum NCIMB 10025 | - |
(3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Corynebacterium glutamicum DSM 20300 | - |
(3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Parageobacillus thermoglucosidasius DSM 2542 | - |
(3S)-acetoin + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | X5F726 | - |
- |
Corynebacterium glutamicum | Q8NMA4 | - |
- |
Corynebacterium glutamicum ATCC 13032 | Q8NMA4 | - |
- |
Corynebacterium glutamicum BCRC 11384 | Q8NMA4 | - |
- |
Corynebacterium glutamicum DSM 20300 | Q8NMA4 | - |
- |
Corynebacterium glutamicum JCM 1318 | Q8NMA4 | - |
- |
Corynebacterium glutamicum LMG 3730 | Q8NMA4 | - |
- |
Corynebacterium glutamicum NCIMB 10025 | Q8NMA4 | - |
- |
Parageobacillus thermoglucosidasius | - |
- |
- |
Parageobacillus thermoglucosidasius DSM 2542 | - |
- |
- |
Pyrococcus furiosus | Q8TZM9 | - |
- |
Pyrococcus furiosus ATCC 43587 | Q8TZM9 | - |
- |
Pyrococcus furiosus JCM 8422 | Q8TZM9 | - |
- |
Pyrococcus furiosus Vc1 | Q8TZM9 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration | Parageobacillus thermoglucosidasius |
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration | Pyrococcus furiosus |
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration | Bacillus subtilis |
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration | Corynebacterium glutamicum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.21 | - |
purified recombinant His-tagged enzyme, pH 8.5, 52°C | Corynebacterium glutamicum |
2.25 | - |
purified recombinant His-tagged enzyme, pH 8.5, 47°C | Bacillus subtilis |
2.25 | - |
purified recombinant His-tagged enzyme, pH 9.0, 67°C | Parageobacillus thermoglucosidasius |
12.74 | - |
purified recombinant His-tagged enzyme, pH 9.0, 72°C | Pyrococcus furiosus |
Storage Stability | Organism |
---|---|
37°C, purified recombinant His-tagged BS-BDH is unstable losing 59% of its initial activity when stored at 37°C for 6 h | Bacillus subtilis |
the purified recombinant His-tagged CG-BDH is very stable when stored below 37°C for 6 h | Corynebacterium glutamicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R,3S)-butane-2,3-diol + NAD+ | - |
Bacillus subtilis | (3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum | (3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum LMG 3730 | (3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum BCRC 11384 | (3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum ATCC 13032 | (3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum JCM 1318 | (3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum NCIMB 10025 | (3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum DSM 20300 | (3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Parageobacillus thermoglucosidasius | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Pyrococcus furiosus | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum LMG 3730 | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum BCRC 11384 | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Pyrococcus furiosus ATCC 43587 | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum ATCC 13032 | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Pyrococcus furiosus Vc1 | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum JCM 1318 | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Pyrococcus furiosus JCM 8422 | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum NCIMB 10025 | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Corynebacterium glutamicum DSM 20300 | (3S)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Parageobacillus thermoglucosidasius DSM 2542 | (3S)-acetoin + NADH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 32000, about, recombinant His-tagged enzyme, SDS-PAGE, x * 27100, about, sequence calculation | Corynebacterium glutamicum |
? | x * 36000, about, recombinant His-tagged enzyme, SDS-PAGE, x * 37300, about, sequence calculation | Pyrococcus furiosus |
? | x * 43000, about, recombinant His-tagged enzyme, SDS-PAGE, x * 37300, about, sequence calculation | Bacillus subtilis |
? | x * 43000, about, recombinant His-tagged enzyme, SDS-PAGE, x * 37900, about, sequence calculation | Parageobacillus thermoglucosidasius |
Synonyms | Comment | Organism |
---|---|---|
BDH | - |
Parageobacillus thermoglucosidasius |
BDH | - |
Pyrococcus furiosus |
BDH | - |
Bacillus subtilis |
BDH | - |
Corynebacterium glutamicum |
BdhA | - |
Pyrococcus furiosus |
BS-BDH | - |
Bacillus subtilis |
CG-BDH | - |
Corynebacterium glutamicum |
Cgl2674 | - |
Corynebacterium glutamicum |
meso-2,3-butanediol dehydrogenase | - |
Parageobacillus thermoglucosidasius |
meso-2,3-butanediol dehydrogenase | - |
Pyrococcus furiosus |
meso-2,3-butanediol dehydrogenase | - |
Bacillus subtilis |
meso-2,3-butanediol dehydrogenase | - |
Corynebacterium glutamicum |
PF-BDH | - |
Pyrococcus furiosus |
PF1960 | - |
Pyrococcus furiosus |
PT-BDH | - |
Parageobacillus thermoglucosidasius |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
47 | - |
- |
Bacillus subtilis |
52 | - |
- |
Corynebacterium glutamicum |
67 | - |
- |
Parageobacillus thermoglucosidasius |
72 | - |
- |
Pyrococcus furiosus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
purified recombinant His-tagged BS-BDH is unstable losing 59% of its initial activity when stored at 37°C for 6 h | Bacillus subtilis |
37 | - |
purified recombinant His-tagged CG-BDH, stable up to, 6 h | Corynebacterium glutamicum |
52 | - |
purified recombinant His-tagged PF-BDH, high resistance to thermal inactivation, 50% activity remaining after 6 h | Pyrococcus furiosus |
52 | - |
purified recombinant His-tagged PT-BDH, highly stable, activity is unchanged after 6 h incubation at 52°C | Parageobacillus thermoglucosidasius |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.59 | - |
NAD+ | pH 8.5, 52°C, recombinant His-tagged enzyme | Corynebacterium glutamicum | |
0.59 | - |
(2R,3S)-butane-2,3-diol | pH 8.5, 52°C, recombinant His-tagged enzyme | Corynebacterium glutamicum | |
2.14 | - |
NAD+ | pH 8.5, 47°C, recombinant His-tagged enzyme | Bacillus subtilis | |
2.14 | - |
(2R,3S)-butane-2,3-diol | pH 8.5, 47°C, recombinant His-tagged enzyme | Bacillus subtilis | |
2.14 | - |
NAD+ | pH 9.0, 67°C, recombinant His-tagged enzyme | Parageobacillus thermoglucosidasius | |
2.14 | - |
(2R,3S)-butane-2,3-diol | pH 9.0, 67°C, recombinant His-tagged enzyme | Parageobacillus thermoglucosidasius | |
11.42 | - |
NAD+ | pH 9.0, 72°C, recombinant His-tagged enzyme | Pyrococcus furiosus | |
11.42 | - |
(2R,3S)-butane-2,3-diol | pH 9.0, 72°C, recombinant His-tagged enzyme | Pyrococcus furiosus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
- |
Bacillus subtilis |
8.5 | - |
- |
Corynebacterium glutamicum |
9 | - |
- |
Parageobacillus thermoglucosidasius |
9 | - |
- |
Pyrococcus furiosus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Parageobacillus thermoglucosidasius | |
NAD+ | - |
Pyrococcus furiosus | |
NAD+ | - |
Bacillus subtilis | |
NAD+ | - |
Corynebacterium glutamicum | |
NADH | - |
Parageobacillus thermoglucosidasius | |
NADH | - |
Pyrococcus furiosus | |
NADH | - |
Bacillus subtilis | |
NADH | - |
Corynebacterium glutamicum |
General Information | Comment | Organism |
---|---|---|
physiological function | D-(-)-acetoin with an optical purity of 25.9% is produced by PT-BDH | Parageobacillus thermoglucosidasius |
physiological function | D-(-)-acetoin with an optical purity of 57% is produced by BS-BDH | Pyrococcus furiosus |
physiological function | L-(+)-acetoin with an optical purity of 92% is produced by BS-BDH | Bacillus subtilis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.077 | - |
(2R,3S)-butane-2,3-diol | pH 8.5, 47°C, recombinant His-tagged enzyme | Bacillus subtilis | |
0.077 | - |
(2R,3S)-butane-2,3-diol | pH 9.0, 67°C, recombinant His-tagged enzyme | Parageobacillus thermoglucosidasius | |
0.219 | - |
(2R,3S)-butane-2,3-diol | pH 9.0, 72°C, recombinant His-tagged enzyme | Pyrococcus furiosus | |
0.227 | - |
(2R,3S)-butane-2,3-diol | pH 8.5, 52°C, recombinant His-tagged enzyme | Corynebacterium glutamicum | |
2.95 | - |
NAD+ | pH 8.5, 52°C, recombinant His-tagged enzyme | Corynebacterium glutamicum | |
17.83 | - |
NAD+ | pH 8.5, 47°C, recombinant His-tagged enzyme | Bacillus subtilis | |
42.8 | - |
NAD+ | pH 9.0, 67°C, recombinant His-tagged enzyme | Parageobacillus thermoglucosidasius | |
57.1 | - |
NAD+ | pH 9.0, 72°C, recombinant His-tagged enzyme | Pyrococcus furiosus |