1.1.1.40: malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+)

This is an abbreviated version, for detailed information about malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+), go to the full flat file.

Reaction

(S)-malate
+
NADP+
=
pyruvate
+
CO2
+
NADPH

Synonyms

ADP-malic enzyme2, c-NADP-ME, C4 NADP-malic enzyme, C4 photosynthetic NADP-malic enzyme, C4-NADP-malic enzyme, C4-NADP-ME, ChlME1, ChlME2, cytoNADPME, cytosolic malic enzyme, cytosolic NADP+-dependent isoform, cytosolic NADP+-dependent malic enzyme, L-malate: NADP oxidoreductase (decarboxylating), L-malate: NADP oxidoreductase [OAA decarboxylating], L-malate: NADP oxidoreductase [oxaloacetate decarboxylating], L-malate:NADP oxidoreductase, L-malate:NADP oxidoreductase (oxaloacetate decarboxylating), MaeB, MalA, malate dehydrogenase (decarboxylating, NADP), malate dehydrogenase (NADP, decarboxylating), malE1, malic enzyme, malic enzyme 1, malic enzyme 3, malic enzyme-NADP, ME, ME-NADP, ME1, ME2, ME3, NAD(P)+-malic enzyme, NADP dependent malic enzyme, NADP malic enzyme, NADP+ dependent malic enzyme, NADP+-dependent malic enzyme, NADP+-dependent malic enzyme 3, NADP+-ME, NADP-dependent malate dehydrogenase, NADP-dependent malic enzyme, NADP-linked decarboxylating malic enzyme, NADP-malate enzyme, NADP-malic enzyme, NADP-malic enzyme 2, NADP-MDH, NADP-ME, NADP-ME1, NADP-ME2, NADP-ME3, NADP-ME4, NADP-specific malate dehydrogenase, NADP-specific malic enzyme, NADP-specific ME, pyruvic-malic carboxylase, TME

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.1 With NAD+ or NADP+ as acceptor
                1.1.1.40 malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+)

Engineering

Engineering on EC 1.1.1.40 - malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+)

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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C99S
-
turnover number decreases 3fold and Km for malate increases 4fold
R70Q
-
kinetic parameters are similar except for a slightly lower turnover number and higher Km for L-malate
D235A
-
turnover number is 783.5fold lower than wild-type value
D257A
-
turnover number is 28.5fold lower than wild-type value
D258A
-
turnover number is 522fold lower than wild-type value
E234A
-
turnover number is 1.3fold higher than wild-type value
K162A
-
site-directed mutagenesis, the mutation does not affect Mn2+ binding of the mutant enzyme, but kcat is 27000fold reduced compared to the wild-type enzyme, NH4Cl shows no rescue of the pyruvate reduction in the K162A mutant, while for oxaloacetate decarboxylation, ammonium chloride demonstrated a maximum restoration of 3.5fold at 1 mM, and its rescue efficiency decreases with increasing concentration
K162Q
-
site-directed mutagenesis, the mutation does not affect Mn2+ binding of the mutant enzyme, but kcat is 3500fold reduced compared to the wild-type enzyme
K162R
-
site-directed mutagenesis,the mutation does not affect Mn2+ binding of the mutant enzyme, but kcat is 125fold reduced compared to the wild-type enzyme
K362A
-
site-directed mutagenesis, 70fold increased Km for NADP+ compared to the wild-type enzyme
W252A
-
site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
W252F
-
site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
W252H
-
site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
W252I
-
site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
W252S
-
site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
Y91F
-
site-directed mutagenesis, the mutation does not affect Mn2+ binding of the mutant enzyme, the mutant shows a 25fold increase and a 3fold decrease in the Km values for (S)-malate and NADP+ respectively, and its kcat value is decreased by 200fold compared to wild-type enzyme
D139A
-
dimeric mutant enzyme with reduced activity compared to the wild type enzyme
D568A
-
dimeric or tetrameric mutant enzyme with increased activity compared to the wild type enzyme
D90A
-
dimeric or tetrameric mutant enzyme with reduced activity compared to the wild type enzyme
E314A
-
site-directed mutagenesis
E314A/S346I/K347D/K362H
-
site-directed mutagenesis, the quadruple mutant enzyme is a mainly NAD+-utilizing enzyme by a considerable increase in catalysis using NAD+ as the cofactor, shows increased inibition by ATP compared to the wild-type enzyme
E314A/S346K
-
site-directed mutagenesis
E314A/S346K/K347Y/K362H
-
site-directed mutagenesis, the quadruple mutant enzyme is a mainly NAD+-utilizing enzyme by a considerable increase in catalysis using NAD+ as the cofactor, shows increased inibition by ATP compared to the wild-type enzyme
E314A/S346K/K347Y/K362Q
-
site-directed mutagenesis, the quadruple mutant enzyme is a mainly NAD+-utilizing enzyme by a considerable increase in catalysis using NAD+ as the cofactor, shows increased inibition by ATP compared to the wild-type enzyme
E73K
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
H142A/D568A
H51A
-
dimeric or tetrameric mutant enzyme with wild type activity
H51A/D139A
H51A/D90A
K347Y
-
site-directed mutagenesis, the mutant enzyme shows a 5fold increased Km for NADP+ compared to the wild-type enzyme
K347Y/K362Q
-
site-directed mutagenesis
K362H
-
site-directed mutagenesis
K362Q
-
site-directed mutagenesis, the mutant enzyme displays a significant, over 140fold elevation in Km,NADP value compared with that of wild-type c-NADP-ME but no significant changes in the kcat,NADP value
N59E
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
N59E/E73K
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
N59E/E73K/S102D
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
S102D
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
S346I/K347D/K362H
-
site-directed mutagenesis, the triple c-NADP-ME mutant does not show significant reduction in its Km,NAD values. This mutant exclusively utilizes NAD+ as its cofactor
S346K
-
site-directed mutagenesis, site-directed mutagenesis, the mutant enzyme shows a 3fold increased Km for NADP+ compared to the wild-type enzyme
S346K/K347Y
-
site-directed mutagenesis, the double mutant enzyme shows a 30fold increased Km for NADP+ compared to the wild-type enzyme
S346K/K347Y/K362H
-
site-directed mutagenesis, the triple c-NADP-ME mutant does not show significant reduction in its Km,NAD values, but displays an enhanced value for kcat,NAD
S346K/K347Y/K362Q
-
site-directed mutagenesis, the triple c-NADP-ME mutant does not show significant reduction in its Km,NAD values
S346K/K362Q
-
site-directed mutagenesis
S57K
the mutant shows increased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
S57K/N59E/E73K
the mutant shows wild type turnover numbers for (S)-malate and NADP+
S57K/N59E/E73K/S102D
the mutant shows increased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
S57K/N59E/E73K/S102D/H74K/D78P/D80E/D87G
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
S57K/N59E/E73K/S102D/H74K/D78P/D80E/D87G/D90E/K106S/Q121S/L125H
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
S57K/N59E/E73K/S102D/H74K/D78P/D80E/D87G/K106S/Q121S/L125H
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
moe
-
enzyme inactivation by shRNA, ME1 activity is reduced by 62% and glucose-induced insulin secretion is decreased
A387G
-
site directed mutagenesis, mutation at the NADP+ binding site, mutant shows 48fold decreased kcat and 4.3 and 5.8fold increased Km for NADP+ and L-malate, respectively, compared to the wild-type enzyme, no activity with NAD+
A392G
-
site directed mutagenesis, mutation at the NADP+ binding site, mutant shows unaltered kcat, but 3.5 and 2.6fold increased Km for NADP+ and L-malate, respectively, and increased activity with NAD+ compared to the wild-type enzyme
C192A
-
marked decrease in kcat value, less than 10% of wild-type, with concomitant increase in Km value for NADP+. Unlike wild-type, activity is not significantly changed in presence of oxidant iodosobenzoate; site-directed mutagenesis of isozyme ZmC4-NADP-ME, the mutation does not affect the tetrameric state, the mutant displays lower malate affinity than the wild-type enzyme
C231A
-
marked decrease in kcat value, less than 10% of wild-type, with concomitant increase in Km value for NADP+. Similar to wild-type, activity decreases in presence of oxidant iodosobenzoate; site-directed mutagenesis of isozyme ZmC4-NADP-ME, the mutation does not affect the tetrameric state, the mutant displays lower malate affinity than the wild-type enzyme
C246A
-
marked decrease in kcat value, less than 10% of wild-type, with concomitant increase in Km value for NADP+. Unlike wild-type, activity is not significantly changed in presence of oxidant iodosobenzoate; site-directed mutagenesis of isozyme ZmC4-NADP-ME, the mutation does not affect the tetrameric state, C246A exhibits a nearly 5fold increase in its affinity towards NADP+ and a 3fold decrease for malate compared to the wild-tpe enzyme
C270A
-
marked decrease in kcat value, less than 10% of wild-type, with concomitant increase in Km value for NADP+. Unlike wild-type, activity is not significantly changed in presence of oxidant iodosobenzoate; site-directed mutagenesis of isozyme ZmC4-NADP-ME, the mutation does not affect the tetrameric state, the mutant displays lower malate affinity than the wild-type enzyme
K225I
-
site-directed mutagenesis, mutation of a conserved residue involved in catalysis and substrate binding, mutant shows highly reduced activity and a 10fold higher partitioning ratio of oxaloacetate and malate compared to the wild-type enzyme, preference for reduction of oxaloacetate instead of decarboxylation
K435L/K436L
-
site-directed mutagenesis, mutation of residues which are important in cofactor binding, over 6fold increased Ki for 2'-AMP, and 1.7fold decreased Ki for 5'-AMP, and increased activity with NAD+ compared to the wild-type enzyme
additional information