Literature summary for 1.1.1.B20 extracted from

Cui, Z.; Mao, Y.; Zhao, Y.; Chen, C.; Tang, Y.; Chen, T.; Ma, H.; Wang, Z.
Concomitant cell-free biosynthesis of optically pure D-(-)-acetoin and xylitol via a novel NAD+ regeneration in two-enzyme cascade (2018), J. Chem. Technol. Biotechnol., 93, 3444-3451 .

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Search for more literature for 1.1.1.B20

Application

Application	Comment	Organism
synthesis	Bacillus subtilis enzyme BS-BDH is a potential candidate for L-(+)-acetoin production	Bacillus subtilis

Cloned(Commentary)

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

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

Inhibitors	Comment	Organism
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 [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
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/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
(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

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 (Commentary)

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 [micromol/min/mg]

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

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(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

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

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 [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
47	-	-	Bacillus subtilis
52	-	-	Corynebacterium glutamicum
67	-	-	Parageobacillus thermoglucosidasius
72	-	-	Pyrococcus furiosus

Temperature Stability [°C]

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 [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
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

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.5	-	-	Bacillus subtilis
8.5	-	-	Corynebacterium glutamicum
9	-	-	Parageobacillus thermoglucosidasius
9	-	-	Pyrococcus furiosus

Cofactor

Cofactor	Comment	Organism
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

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 [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
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