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2,4'-dichloroacetophenone + NADH + H+
? + NAD+
-
Substrates: binding and transformation of unnatural 2,4-dichloroacetophenone is not as for good as natural substrates, although it is reduced with very high catalytic efficiency
Products: -
?
2-deoxy-D-galactose + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
2-deoxy-D-ribose + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
9,10-phenanthrenequinone + NADPH + H+
? + NADP+
Substrates: -
Products: -
?
butanal + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
D-arabinose + NADPH + H+
D-arabinitol + NADP+
D-erythrose + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
D-erythrose + NADPH + H+
D-erythritol + NADP+
D-fucose + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
D-galactose + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
D-glucose + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
D-glucose + NADPH + H+
? + NADP+
-
Substrates: -
Products: -
?
D-lyxose + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
D-ribose + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
D-xylose + NAD(P)H + H+
xylitol + NAD(P)+
-
Substrates: -
Products: -
?
D-xylose + NADH + H+
xylitol + NAD+
D-xylose + NADPH + H+
xylitol + NADP+
DL-glyceraldehyde + NADH + H+
glycerol + NAD+
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
L-arabinose + NADH + H+
L-arabinitol + NAD+
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
L-lyxose + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
pentanal + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
propionaldehyde + NADH + H+
?
-
Substrates: dual specific xylose reductase (dsXR)
Products: -
?
additional information
?
-
D-arabinose + NADPH + H+
D-arabinitol + NADP+
Substrates: -
Products: -
?
D-arabinose + NADPH + H+
D-arabinitol + NADP+
Substrates: -
Products: -
?
D-erythrose + NADPH + H+
D-erythritol + NADP+
Substrates: -
Products: -
?
D-erythrose + NADPH + H+
D-erythritol + NADP+
Substrates: -
Products: -
?
D-ribose + NADPH + H+
?
Substrates: -
Products: -
?
D-ribose + NADPH + H+
?
Substrates: -
Products: -
?
D-xylose + NADH + H+
xylitol + NAD+
Substrates: the enzyme specifically transfers the 4-pro-R hydrogen from the C-4 of the nicotinamide ring to the re face of the carbonyl carbon of the substrate
Products: -
r
D-xylose + NADH + H+
xylitol + NAD+
Substrates: the enzyme specifically transfers the 4-pro-R hydrogen from the C-4 of the nicotinamide ring to the re face of the carbonyl carbon of the substrate
Products: -
r
D-xylose + NADH + H+
xylitol + NAD+
-
Substrates: wild-type enzyme prefers NADPH over NADH
Products: -
?
D-xylose + NADH + H+
xylitol + NAD+
-
Substrates: -
Products: -
?
D-xylose + NADH + H+
xylitol + NAD+
Substrates: -
Products: -
r
D-xylose + NADH + H+
xylitol + NAD+
-
Substrates: aldehyde reduction is favoured
Products: -
r
D-xylose + NADH + H+
xylitol + NAD+
-
Substrates: using a modified iterative protein redesign and optimization workflow, a sets of mutations is identified that change the nicotinamide cofactor specificity of xylose reductase (CbXR) from its physiological preference for NADPH, to the alternate cofactor NADH
Products: -
?
D-xylose + NADH + H+
xylitol + NAD+
-
Substrates: reaction is catalyzed by dual specific xylose reductase (dsXR), reaction is not catalyzed by NADPH-dependent monospecific xylose reductase (msXR)
Products: -
?
D-xylose + NADPH + H+
xylitol + NADP+
Substrates: -
Products: -
?
D-xylose + NADPH + H+
xylitol + NADP+
Substrates: -
Products: -
?
D-xylose + NADPH + H+
xylitol + NADP+
-
Substrates: xylose reductase is one of the key enzymes for xylose fermentation
Products: -
?
D-xylose + NADPH + H+
xylitol + NADP+
-
Substrates: wild-type enzyme prefers NADPH over NADH
Products: -
?
D-xylose + NADPH + H+
xylitol + NADP+
Substrates: -
Products: -
r
D-xylose + NADPH + H+
xylitol + NADP+
Substrates: -
Products: -
?
D-xylose + NADPH + H+
xylitol + NADP+
Substrates: -
Products: -
r
D-xylose + NADPH + H+
xylitol + NADP+
-
Substrates: using a modified iterative protein redesign and optimization workflow, a sets of mutations is identified that change the nicotinamide cofactor specificity of xylose reductase (CbXR) from its physiological preference for NADPH, to the alternate cofactor NADH
Products: -
?
D-xylose + NADPH + H+
xylitol + NADP+
-
Substrates: reaction is catalyzed by NADPH-dependent monospecific xylose reductase (msXR), and by dual specific xylose reductase (dsXR)
Products: -
?
additional information
?
-
-
Substrates: substrates preferentially bind as alpha-anomers of the pyranose forms. The alpha-anomers are transformed faster, predominately leading to saturation transfer difference effects in the formed products, and can be better docked into the active site than the beta-anomer. The reduction is initiated by alpha-xylopyranose ring opening prior to hydride transfer from NADH
Products: -
?
additional information
?
-
-
Substrates: Candida intermedia produces two isoforms of xylose reductase: one is NADPH-dependent (monospecific xylose reductase, msXR, cf. 1.1.1.431), and another prefers NADH about 4fold over NADPH (dual specific xylose reductase, dsXR)
Products: -
-
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NAD+
-
-
NADH
-
-
NADH
-
dual specific xylose reductase (dsXR) has an about 4fold higher specificity for NADH than NADPH
NADH
strongly prefers NADH to NADPH
NADH
-
transient-state and steady-state kinetic studies of the mechanism of NADH-dependent aldehyde reduction
NADH
-
using a modified iterative protein redesign and optimization workflow, a sets of mutations is identified that change the nicotinamide cofactor specificity of xylose reductase (CbXR) from its physiological preference for NADPH, to the alternate cofactor NADH
NADH
-
wild-type enzyme prefers NADPH over NADH. Mutant enzyme K270S/N272P/S271G/R276F shows a 25fold preference toward NADH over NADPH by a factor of about 13fold, or an improvement of about 42fold, as measured by the ratio of the specificity constant kcat/Km coenzyme
NADP+
-
-
NADPH
-
NADPH
-
preferred cofactor
NADPH
-
dual specific xylose reductase (dsXR) has an about 4fold higher specificity for NADH than NADPH. NADPH-dependent monospecific xylose reductase (msXR) shows non activity with NADH
NADPH
strongly prefers NADH to NADPH
NADPH
-
using a modified iterative protein redesign and optimization workflow, a sets of mutations is identified that change the nicotinamide cofactor specificity of xylose reductase (CbXR) from its physiological preference for NADPH, to the alternate cofactor NADH
NADPH
-
wild-type enzyme prefers NADPH over NADH. Mutant enzyme K270S/N272P/S271G/R276F shows a 25fold preference toward NADH over NADPH by a factor of about 13fold, or an improvement of about 42fold, as measured by the ratio of the specificity constant kcat/Km coenzyme
additional information
-
Kluyveromyces marxianus strains expressing Pichia stipitis Psxyl1 genes show reversed cofactor specificity, overview
-
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
-
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0.126
2-deoxy-D-galactose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
33
Butanal
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.007
D-fucose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.015
D-galactose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.033
D-glucose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
2.43
DL-glyceraldehyde
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.028
L-arabinose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.144
L-Lyxose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.027
NAD+
-
pH 7.0, 25°C
14.7
pentanal
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
13.2
propionaldehyde
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.02
D-erythrose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.068
D-ribose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.01
D-xylose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
29.8
D-xylose
pH 6.5, 42°C, recombinant His-tagged enzyme
31.5
D-xylose
pH 6.0, coenzyme: NADH
78
D-xylose
-
pH 7.0, 25°C
82
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADPH, wild-type enzyme
90
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADH, wild-type enzyme
168
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADPH, mutant enzyme K270S/N272P/S271G/R276F
244.3
D-xylose
pH 6.0, coenzyme: NADPH
291
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADH, mutant enzyme K270S/N272P/S271G/R276F
0.0033
NADH
pH 6.0
0.0106
NADH
-
pH 6.0, temperature not specified in the publication, wild-type enzyme, wild-type enzyme
0.015
NADH
-
pH 7.0, 25°C
0.04
NADH
pH 6.5, 42°C, recombinant His-tagged enzyme
0.147
NADH
-
pH 6.0, temperature not specified in the publication, mutant enzyme K270S/N272P/S271G/R276F
0.0062
NADPH
-
pH 6.0, temperature not specified in the publication, wild-type enzyme, wild-type enzyme
0.427
NADPH
-
pH 6.0, temperature not specified in the publication, mutant enzyme K270S/N272P/S271G/R276F
209
xylitol
-
pH 7.0, 25°C
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3.5
2-deoxy-D-galactose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.027 - 12
9,10-phenanthrenequinone
5.4
Butanal
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
27.5
D-erythrose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
10.2
D-fucose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
9.4
D-galactose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
5.6
D-glucose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
4.9
D-ribose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
14.1
DL-glyceraldehyde
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
13.5
L-arabinose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
6.6
L-Lyxose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
5.9
pentanal
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
4.6
propionaldehyde
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.92
xylitol
-
pH 7.0, 25°C
0.027
9,10-phenanthrenequinone
mutant K80A, pH 7.0, 25°C
0.043
9,10-phenanthrenequinone
mutant H113A, pH 7.0, 25°C
0.2
9,10-phenanthrenequinone
mutant Y51A, pH 7.0, 25°C
12
9,10-phenanthrenequinone
wild-type, pH 7.0, 25°C
0.002
D-xylose
mutant K80A, pH 7.0, 25°C
0.02
D-xylose
mutant H113A, pH 7.0, 25°C
2.6
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADPH, mutant enzyme K270S/N272P/S271G/R276F
10
D-xylose
wild-type, pH 7.0, 25°C
11.42
D-xylose
pH 6.5, 42°C, recombinant His-tagged enzyme
12
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADH, mutant enzyme K270S/N272P/S271G/R276F
14.2
D-xylose
-
pH 7.0, 25°C
15.4
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADH, wild-type enzyme
16.9
D-xylose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
27.5
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADPH, wild-type enzyme
11.42
NADH
pH 6.5, 42°C, recombinant His-tagged enzyme
12
NADH
-
pH 6.0, temperature not specified in the publication, mutant enzyme K270S/N272P/S271G/R276F
15.4
NADH
-
pH 6.0, temperature not specified in the publication, wild-type enzyme, wild-type enzyme
2.6
NADPH
-
pH 6.0, temperature not specified in the publication, mutant enzyme K270S/N272P/S271G/R276F
27.5
NADPH
-
pH 6.0, temperature not specified in the publication, wild-type enzyme, wild-type enzyme
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28
2-deoxy-D-galactose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
20 - 2300
9,10-phenanthrenequinone
0.16
Butanal
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
1457
D-fucose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
627
D-galactose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
170
D-glucose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
5.8
DL-glyceraldehyde
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
482
L-arabinose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
46
L-Lyxose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.4
pentanal
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.035
propionaldehyde
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
20
9,10-phenanthrenequinone
mutant H113A, pH 7.0, 25°C
33
9,10-phenanthrenequinone
mutant Y51A, pH 7.0, 25°C
500
9,10-phenanthrenequinone
mutant K80A, pH 7.0, 25°C
2300
9,10-phenanthrenequinone
wild-type, pH 7.0, 25°C
0.089
D-erythrose
pH 6.0
1380
D-erythrose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.043
D-ribose
pH 6.0
72
D-ribose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
0.0009
D-xylose
mutant K80A, pH 7.0, 25°C
0.019
D-xylose
pH 6.0, coenzyme: NADPH
0.18
D-xylose
-
pH 7.0, 25°C
0.2
D-xylose
mutant H113A, pH 7.0, 25°C
0.383
D-xylose
pH 6.5, 42°C, recombinant His-tagged enzyme
1.5
D-xylose
pH 6.0, coenzyme: NADH
6.2
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADPH, mutant enzyme K270S/N272P/S271G/R276F
81.7
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADH, mutant enzyme K270S/N272P/S271G/R276F
1460
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADH, wild-type enzyme
1690
D-xylose
-
pH 7.0, 25°C, dual specific xylose reductase, cofactor: NADH
4648
D-xylose
-
pH 6.0, temperature not specified in the publication, cofactor: NADPH, wild-type enzyme
81.7
NADH
-
pH 6.0, temperature not specified in the publication, mutant enzyme K270S/N272P/S271G/R276F
285.5
NADH
pH 6.5, 42°C, recombinant His-tagged enzyme
946.7
NADH
-
pH 7.0, 25°C
1460
NADH
-
pH 6.0, temperature not specified in the publication, wild-type enzyme, wild-type enzyme
6.2
NADPH
-
pH 6.0, temperature not specified in the publication, mutant enzyme K270S/N272P/S271G/R276F
4648
NADPH
-
pH 6.0, temperature not specified in the publication, wild-type enzyme, wild-type enzyme
0.0044
xylitol
-
pH 7.0, 25°C
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K21A
-
mutation reverses the cofactor specificity from major NADP- to NAD-dependent
K270N
-
mutation reverses the cofactor specificity from major NADP- to NAD-dependent
K270S/N272P/S271G/R276F
-
the mutant shows a 25fold preference toward NADH over NADPH by a factor of about 13fold, or an improvement of about 42fold, as measured by the ratio of the specificity constant kcat/Km coenzyme. Compared with the wild-type, the kcat(NADH) is slightly lower, while the kcat(NADPH) decreases by a factor of about 10
H113A
mutation causes a 10000-100000fold decrease in the rate constant for hydride transfer from NADH to 9,10-phenanthrenequinone, whose value in the wild-type enzyme is about 800 per s
L80A
mutation causes a 10000-100000fold decrease in the rate constant for hydride transfer from NADH to 9,10-phenanthrenequinone, whose value in the wild-type enzyme is about 800 per s
Y51A
mutation causes a 10000-100000fold decrease in the rate constant for hydride transfer from NADH to 9,10-phenanthrenequinone, whose value in the wild-type enzyme is about 800 per s
K274R
-
mutation introduced to change the specificity toward NADH. Fermentation with the mutant strain shows a 2.8fold reduction in xylitol accumulation and 4.5fold increase in citric acid production compared to the wild-type strain
K274R
-
mutation introduced to change the specificity toward NADH. Fermentation with the mutant strain shows a 2.8fold reduction in xylitol accumulation and 4.5fold increase in citric acid production compared to the wild-type strain
-
K274R/N276D
-
structure-guided site-directed mutagenesis, change of the coenzyme preference of the xyluose reductase about 170fold from NADPH in the wild-type to NADH, which, in spite of the structural modifications introduced, has retained the original catalytic efficiency for reduction of xylose by NADH
K274R/N276D
NADH-specific mutant, Saccharomyces cerevisiae expressing mutant K274R/N276D exhibits intracellular activities of 0.94 U/mg 1.07 U/mg with NADPH and NADH, respectively
additional information
-
the native xylose reductase gene Kmxyl1 of the Kluyveromyces marxianus strain YHJ010 is substituted with xylose reductase or its mutants N272D, K21A/N272D, or K270M from Pichia stipitis, i.e. Scheffersomyces stipitis, resulting in Kluyveromyces marxianus strains YZB013, YZB014, and YZB015. The ability of the resultant recombinant strains to assimilate xylose to produce xylitol and ethanol at elevated temperature is greatly improved, overview. But the strain YZB015 expressing a mutant PsXR K21A/N272D, with which co-enzyme preference is completely reversed from NADPH to NADH, fails to ferment due to the low expression
additional information
-
the native xylose reductase gene Kmxyl1 of the Kluyveromyces marxianus strain YHJ010 is substituted with xylose reductase or its mutants N272D, K21A/N272D, or K270M from Pichia stipitis, i.e. Scheffersomyces stipitis, resulting in Kluyveromyces marxianus strains YZB013, YZB014, and YZB015. The ability of the resultant recombinant strains to assimilate xylose to produce xylitol and ethanol at elevated temperature is greatly improved, overview. But the strain YZB015 expressing a mutant PsXR K21A/N272D, with which co-enzyme preference is completely reversed from NADPH to NADH, fails to ferment due to the low expression
-
additional information
the combinatorial expression of two xylose reductase (XR) genes and two xylitol dehydrogenase (XDH) genes from Spathaspora passalidarum and the heterologous expression of the Piromyces sp. xylose isomerase (XI) gene are induced in Aureobasidium pullulans strain CBS 110374. Overexpression of XYL1.2 (encoding XR) and XYL2.2 (encoding XDH) is the most beneficial for xylose utilization, resulting in a 17.76% increase in consumed xylose compared with the parent strain, whereas the introduction of the Piromyces sp. XI pathway fails to enhance xylose utilization efficiency. Construction of knock-in mutants, method, evaluation of internal redox state, xylose utilization and cell growth of the recombinant strains, pH 5.8, 28°C, comparison the fermentation abilities of the parent strain and the recombinant strains using several carbon sources, overview
additional information
the combinatorial expression of two xylose reductase (XR) genes and two xylitol dehydrogenase (XDH) genes from Spathaspora passalidarum and the heterologous expression of the Piromyces sp. xylose isomerase (XI) gene are induced in Aureobasidium pullulans strain CBS 110374. Overexpression of XYL1.2 (encoding XR) and XYL2.2 (encoding XDH) is the most beneficial for xylose utilization, resulting in a 17.76% increase in consumed xylose compared with the parent strain, whereas the introduction of the Piromyces sp. XI pathway fails to enhance xylose utilization efficiency. Construction of knock-in mutants, method, evaluation of internal redox state, xylose utilization and cell growth of the recombinant strains, pH 5.8, 28°C, comparison the fermentation abilities of the parent strain and the recombinant strains using several carbon sources, overview
additional information
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the combinatorial expression of two xylose reductase (XR) genes and two xylitol dehydrogenase (XDH) genes from Spathaspora passalidarum and the heterologous expression of the Piromyces sp. xylose isomerase (XI) gene are induced in Aureobasidium pullulans strain CBS 110374. Overexpression of XYL1.2 (encoding XR) and XYL2.2 (encoding XDH) is the most beneficial for xylose utilization, resulting in a 17.76% increase in consumed xylose compared with the parent strain, whereas the introduction of the Piromyces sp. XI pathway fails to enhance xylose utilization efficiency. Construction of knock-in mutants, method, evaluation of internal redox state, xylose utilization and cell growth of the recombinant strains, pH 5.8, 28°C, comparison the fermentation abilities of the parent strain and the recombinant strains using several carbon sources, overview
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)
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synthesis
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D-xylose is the second most abundant renewable sugar in nature, and its fermentation to ethanol has great economical potential. Unfortunately, Saccharomyces cerevisiae, which has been optimized for ethanol production, cannot utilize xylose efficiently, while D-xylulose, an isomerization product of D-xylose, can be assimilated. A major strategy for constructing xylose-fermenting Saccharomyces cerevisiae is to introduce genes involved in xylose metabolism from other organisms. Xylose reductase and xylitol dehydrogenase (EC 1.1.1.9) from the xylose-fermenting yeast Pichia stipitis are cloned into Saccharomyces cerevisiae to allow xylose fermentation to ethanol. In this case, xylose is converted into xylulose by the sequential actions of two oxidoreductases. First, Pichia stipitis xylose reductase catalyses the reduction of xylose into xylitol with NAD(P)H as co-substrate. Xylitol is then oxidized by PsXDH (Pichia stipitis xylitol dehydrogenase) which uses NAD+ exclusively as co-substrate to yield xylulose. The different coenzyme specificity of the two enzymes xylose reductase and xylitol dehydrogenase, however, creates an intracellular redox imbalance, which results in low ethanol yields and considerable xylitol by-product formation. A mutant is constructed that shows an altered active site that is more unfavorable for NADPH than NADH in terms of both Km and kcat. There are potentials for application of the mutant (K270S/N272P/S271G/R276F) in constructing a more balanced xylose reductase/xylitol dehydrogenase pathway in recombinant xylose-fermenting Saccharomyces cerevisiae strains
synthesis
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dual specific xylose reductase (dsXR) has an about 4fold higher specificity for NADH than NADPH. This fact could make this enzyme an interesting candidate to be used in metabolic engineering of the yeast xylose metabolism, likely in Saccharomyces cerevisiae. Increased levels of dsXR activity could contribute to an improvement of ethanol production from D-xylose by reducing the cofactor imbalance of the initial catabolic pathway
synthesis
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fermentation of mixed glucose-xylose substrates in Saccharomyces cerevisiae strains BP10001 and BP000, expressing Candida tenuis xylose reductase in mutated NADH-preferring form and NADPH-preferring wild-type form, respectively. Glucose and xylose, each at 10 g/l, are converted sequentially. The distribution of fermentation products from glucose is identical for both strains whereas when using xylose, BP10001 shows enhanced ethanol yield and decreased yields of xylitol and glycerol as compared to BP000. Increase in xylose concentration from 10 to 50 g/l results in acceleration of substrate uptake by BP10001 and reduction of the xylitol yield. In mixed substrate batches, xylose is taken up at low glucose concentrations and up to 5fold enhanced xylose uptake rate is found towards glucose depletion
synthesis
expression of wild-type Xyl1 or an NADH-specific mutant in Saccharomyces cerevisiae. The Xyl1 mutant decreases the biocatalysts performance, suggesting use of the NADPH-preferring wild-type enzyme when (semi-)aerobic conditions are applied. In a bioreactor process, the best-performing strain converts 40 g/l xylose with an initial productivity of 1.16 g/l/h and a xylitol yield of 100%
synthesis
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in fermentation for citric acid production and xylitol accumulation by using D-xylose as the sole carbon source, a sttrain carrying mutant K274R shows a 2.8fold reduction in xylitol accumulation and 4.5fold increase in citric acid production compared to the wild-type strain
synthesis
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in fermentation for citric acid production and xylitol accumulation by using D-xylose as the sole carbon source, a sttrain carrying mutant K274R shows a 2.8fold reduction in xylitol accumulation and 4.5fold increase in citric acid production compared to the wild-type strain
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Lee, J.K.; Koo, B.S.; Kim, S.Y.
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Yamadazyma tenuis (O74237)
brenda
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2013
Yamadazyma tenuis
-
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93
3444-3451
2018
Yamadazyma tenuis (O74237)
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brenda
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Spathaspora passalidarum (A0A0S2PJI4), Spathaspora passalidarum (S4WCW2), Spathaspora passalidarum
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Scheffersomyces stipitis
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