1.1.1.86: ketol-acid reductoisomerase (NADP+)

This is an abbreviated version, for detailed information about ketol-acid reductoisomerase (NADP+), go to the full flat file.

Reaction

(2R)-2,3-dihydroxy-3-methylbutanoate
+
NADP+
=
(2S)-2-hydroxy-2-methyl-3-oxobutanoate
+
NADPH
+
H+

Synonyms

2-hydroxy-3-keto acid reductoisomerase, acetohydroxy acid isomeroreductase, acetohydroxy acid reductoisomerase, acetohydroxy-acid isomeroreductase , acetohydroxy-acid reductoisomerase , acetohydroxyacid isomeroreductase, acetolactate reductoisomerase, AHAIR, AHIR, alpha-keto-beta-hydroxylacil reductoisomerase, alpha-keto-beta-hydroxylacyl reductoisomerase, class II ketol-acid reductoisomerase, dehydrogenase, dihydroxyisovalerate (isomerizing), dihydroxyisovalerate dehydrogenase (isomerizing), Icl1p, Ilv5p, ilvC, IlvC-PanE, IlvC1, IlvC2, isomerase, ketol acid reducto-, isomeroreductase, KARI, ketol-acid reductoisomerase, reductoisomerase

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.1 With NAD+ or NADP+ as acceptor
                1.1.1.86 ketol-acid reductoisomerase (NADP+)

Engineering

Engineering on EC 1.1.1.86 - ketol-acid reductoisomerase (NADP+)

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
R48P/S51L/S52D
-
mutant engineered for reversal in cofactor specificity for NADH over NADPH
R48P/S51L/S52D/R84A
-
mutant engineered for reversal in cofactor specificity for NADH over NADPH
R48P/S51L/S52D
-
mutant engineered for reversal in cofactor specificity for NADH over NADPH
-
R48P/S51L/S52D/R84A
-
mutant engineered for reversal in cofactor specificity for NADH over NADPH
-
D217E
-
less than 4% reductoisomerase activity in comparison to wild-type enzyme
D217N
-
less than 4% reductoisomerase activity in comparison to wild-type enzyme
E213D
-
75% reductoisomerase activity in comparison of wild-type enzyme
E213Q
-
less than 4% reductoisomerase activity in comparison of wild-type enzyme
E221D
-
less than 4% reductoisomerase activity in comparison to wild-type enzyme
E221Q
-
less than 4% reductoisomerase activity in comparison to wild-type enzyme
E389D
-
less than 4% reductoisomerase activity in comparison to wild-type enzyme
E389Q
-
less than 4% reductoisomerase activity in comparison to wild-type enzyme
E393D
-
less than 4% reductoisomerase activity in comparison to wild-type enzyme
E393Q
-
the mutant is insoluble, a soluble form is obtained only after denaturation
H132K
-
less than 4% reductoisomerase activity in comparison of wild-type enzyme
H132Q
-
less than 4% reductoisomerase activity in comparison of wild-type enzyme
K155E
-
less than 4% reductoisomerase activity in comparison of wild-type enzyme
K155Q
-
less than 4% reductoisomerase activity in comparison of wild-type enzyme
K155R
-
less than 4% reductoisomerase activity in comparison of wild-type enzyme
K69L
-
Km-value for NADP+ in the reaction with 2,3-dihydroxy-3-methylbutanoate is 2.1fold higher than the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 163 compared to wild-type enzyme. Km-value for NADPH in the reaction with acetolactate is comparable to that of the wild-type emzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is 1.8fold higher than that of the wild-type enzyme
K75Q
-
Km-value for NADP+ in the reaction with 2,3-dihydroxy-3-methylbutanoate is 2.7fold higher than the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 77.5 compared to wild-type enzyme. Km-value for NADPH in the reaction with acetolactate is lower by a factor 2.9 compared to the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 12.9 compared to wild-type enzyme
R68D/K69L/K75V/R76D
-
turnover-number for reaction with NADH and acetolactate is 48fold higher compared to wild-type enzyme, turnover-number for reaction with NADPH and acetolactate is lower by factor 3.7 compared to wild-type enzyme, Km-value for NADH in the reaction with NADH and acetolactate is lower by a factor 10.8 compared to wild-type enzyme, Km-value for NADH in the reaction with NADPH and acetolactate is 30fold higher compared to wild-type enzyme
R68Q
-
Km-value for NADP+ in the reaction with 2,3-dihydroxy-3-methylbutanoate is 6.9fold higher than the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 345 compared to wild-type enzyme. Km-value for NADPH in the reaction with acetolactate is 3.4fold higher thahn that of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 18 compared to wild-type enzyme
R76D
-
turnover-number for reaction with NADH and acetolactate is 48fold higher compared to wild-type enzyme, turnover-number for reaction with NADPH and acetolactate is lower by factor 4 compared to wild-type enzyme, Km-value for NADH in the reaction with NADH and acetolactate is lower by a factor 2.5 compared to wild-type enzyme, Km-value for NADPH in the reaction with NADPH and acetolactate is 55fold higher compared to wild-type enzyme
R76Q
-
Km-value for NADP+ in the reaction with 2,3-dihydroxy-3-methylbutanoate is 17.1fold higher than the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 258 compared to wild-type enzyme. Km-value for NADPH in the reaction with acetolactate is 5fold higher than that of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 19.5 compared to wild-type enzyme
R76Q/R68A
-
turnover-number for reaction with NADH and acetolactate is 20fold higher compared to wild-type enzyme, turnover-number for reaction with NADPH and acetolactate is lower by factor 28 compared to wild-type enzyme, Km-value for NADH in the reaction with NADH and acetolactate is comparable to that of wild-type enzyme, Km-value for NADPH in the reaction with NADPH and acetolactate is 22fold higher compared to wild-type enzyme
S414A
-
less than 4% reductoisomerase activity in comparison to wild-type enzyme
S414T
-
less than 4% reductoisomerase activity in comparison to wild-type enzyme
G50D/S52D
DELTA1-17
-
mutant with N-terminal 17-residue deletion: cellular localisation similar to wild-type, introduction into an ilv5DELTA strain, lacking the ilv5 gene, complements isoleucine and valine synthesis
DELTA1-33
-
mutant with N-terminal 33-residue deletion: cellular localisation predominatly in the cytosol rather than in mitochondria. Deletion of the N-terminal 33 residues is sufficient to largely impair the protein's targeting to mitochondria. Introduction into an ilv5DELTA strain, lacking the ilv5 gene, complements isoleucine and valine synthesis
DELTA1-40
-
mutant with N-terminal 40-residue deletion: cellular localisation throughout the cytosol rather than in mitochondria
DELTA1-46
-
mutant with N-terminal 46-residue deletion: cellular localisation throughout the cytosol rather than in mitochondria. This mutant shows the largest steady-state protein level. Introduction into an ilv5DELTA strain, lacking the ilv5 gene, does not complements isoleucine and valine synthesis. Introduction into an industrial lager brewing strain, a robust expression of DELTA46 is as effective as that of a wild-type Ilv5p in lowering the total Vicinal diketones production in a 2l scale fermentation trial. Additional expression of DELTA46 does not alter the quality of the resultant beer in terms of contents of aromatic compounds and organic acids
DELTA1-53
-
mutant with N-terminal 53-residue deletion: cellular localisation throughout the cytosol rather than in mitochondria
DELTA1-66
-
mutant with N-terminal 66-residue deletion: weaker cytosolic localisation compared to other mutants. Mutant shows only a small amount of steady-state protein content
DELTA1-83
-
mutant with N-terminal 83-residue deletion: weaker cytosolic localisation compared to other mutants. Mutant shows only a small amount of steady-state protein content
DELTA1-99
-
mutant with N-terminal 99-residue deletion: weaker cytosolic localisation compared to other mutants. Mutant shows only a small amount of steady-state protein content
DELTA423-430/F431S
-
mutant enzyme behaves as an active monomer with reduced thermal stability, KM-value for NADPH does not differ considerably from that for the wild-type enzyme, magnesium affinity is dramatically altered by monomerization