1.1.99.31: (S)-mandelate dehydrogenase
This is an abbreviated version!
For detailed information about (S)-mandelate dehydrogenase, go to the full flat file.
Word Map on EC 1.1.99.31
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1.1.99.31
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s-mandelate
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putida
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benzoylformate
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fmn-dependent
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benzaldehyde
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calcoaceticus
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fmn
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mitra
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alpha-hydroxy
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l-lactate
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phenylglyoxylate
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carbanion
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flavocytochrome
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membrane-binding
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flavoenzyme
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alpha-protons
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half-reaction
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rhodotorula
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graminis
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electron-rich
- 1.1.99.31
- s-mandelate
- putida
- benzoylformate
-
fmn-dependent
- benzaldehyde
- calcoaceticus
- fmn
- mitra
-
alpha-hydroxy
- l-lactate
- phenylglyoxylate
-
carbanion
-
flavocytochrome
-
membrane-binding
-
flavoenzyme
-
alpha-protons
-
half-reaction
-
rhodotorula
- graminis
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electron-rich
Reaction
Synonyms
(S)-mandelate dehydrogenase, L-mandelate dehydrogenase, L-MDH, MDH, SManDH, SMDH
ECTree
1.1.99.31 (S)-mandelate dehydrogenaseAdvanced search results
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results (Engineering
Engineering on EC 1.1.99.31 - (S)-mandelate dehydrogenase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
G81A
G81D
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extreme low activity, reduction in activity is due to the decrease in electrophilicity of the FMN
G81S
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the rate of the first half-reaction is slower than the wild-type rate. The rate of the first half-reaction is slower than the wild-type rate. Affinity for O2 increases 10-15fold
G81V
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extreme low activity, reduction in activity is due to the decrease in electrophilicity of the FMN
H274A
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inactive mutant, activity can partially be restored by the addition of exogenous imidazole
H274G
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inactive mutant, activity can be restored by the addition of exogenous imidazole
R165E
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mutant is less stable than wild-type enzyme, mutant enzyme is only partially reduced by (S)-mandelate. 81.8fold decrease in kcat for (S)-mandelate, 199fold increase in KM-value for (S)-mandelate compared to wild-type enzyme
R165G
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mutant is less stable than wild-type enzyme, mutant enzyme is only partially reduced by (S)-mandelate. 27.3fold decrease in kcat for (S)-mandelate, 48.3fold increase in KM-value for (S)-mandelate compared to wild-type enzyme
R165G/R277G
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no activity. Double mutant is less stable than single Arg165 mutant in term of long-term storage
R165G/R277K
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double mutant is less stable than single Arg165 mutant in term of long-term storage. 9000fold decrease in kcat for (S)-mandelate, 261.7fold increase in KM-value for (S)-mandelate compared to wild-type enzyme
R165K
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mutant is less stable than wild-type enzyme, mutant enzyme is fully reduced on addition of (S)-mandelate. 3fold decrease in kcat for (S)-mandelate, 12.5fold increase in KM-value for (S)-mandelate compared to wild-type enzyme
R165K/R277G
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no activity. Double mutant is less stable than single Arg165 mutant in term of long-term storage
R165K/R277K
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double mutant is less stable than single Arg165 mutant in term of long-term storage. 327fold decrease in kcat for (S)-mandelate, 126.7fold increase in KM-value for (S)-mandelate compared to wild-type enzyme
R165M
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mutant is less stable than wild-type enzyme, mutant enzyme is fully reduced on addition of (S)-mandelate. 19.6fold decrease in kcat for (S)-mandelate, 36.7fold increase in KM-value for (S)-mandelate compared to wild-type enzyme
R277G
R277H
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kcat for (S)-mandelate is 1000fold lower than wild-type value, KM-value for (S)-mandelate is 608fold higher than wild-type value
R277K
R277L
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kcat for (S)-mandelate is 421fold lower fold than wild-type value, KM-value for (S)-mandelate is 392fold higher than wild-type value
additional information
G81A
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higher specificity for small substrates, compared to that of wild-type enzyme, the affinity for (S)-mandelate is relatively unchanged. The rate of the first half-reaction is slower than the wild-type rate. 2fold increase in oxidative half-reaction. Affinity for oxygen increases 10-15fold
G81A
mutation in the catalytically similar chimera MDH-GOX2 (mandelate dehydrogenase/glycolate oxidase), mutant has lower reactivity with substrate
R277G
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1565fold decrease in kcat for (S)-mandelate, 141fold increase in KM-value for (S)-mandelate compared to wild-type enzyme
R277G
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kcat for (S)-mandelate is 178fold lower than wild-type value, KM-value for (S)-mandelate is 160fold higher than wild-type value, KM-value for {(S)-3-phenyllactate is 1.6fold lower than wild-type value
R277G
reduced activity compared to the wild type enzyme, the mutation at Arg277 has no influence on the binding affinity of ester substrates
R277K
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5.5 fold decrease in kcat for (S)-mandelate, 46.7fold increase in KM-value for (S)-mandelate compared to wild-type enzyme
R277K
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kcat for (S)-mandelate is 3.7fold lower than wild-type value, KM-value for (S)-mandelate is 36.7fold higher than wild-type value
R277K
reduced activity compared to the wild type enzyme, the mutation at Arg277 has no influence on the binding affinity of ester substrates
additional information
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chimeric mutant of (S)-mandelate dehydrogenase with membrane anchoring loop replaced by a portion of glycolate oxidase from spinach is soluble and retains partial catalytic activity (about 1%) using (S)-mandelate as substrate. The activities of the soluble mutant enzymes (S)-mandelate dehydrogenase with residues 2-4 deleted and (S)-mandelate dehydrogenase with 17 residues deleted at the carboxy terminus are nearly the same when (S)-phenyllactate is used as substrate
additional information
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MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach. G81A/MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach and a G81A mutation in MDH
additional information
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MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach. This mutant is very similar to the wild-type membrane-bound enzyme in its spectroscopic properties, substrate specificity, catalytic activity, kinetic mechanism, and lack of reactivity toeards oxygen. It should prove to be a highly useful model for structural studies of MDH
