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C99S
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turnover number decreases 3fold and Km for malate increases 4fold
R70Q
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kinetic parameters are similar except for a slightly lower turnover number and higher Km for L-malate
D235A
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turnover number is 783.5fold lower than wild-type value
D257A
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turnover number is 28.5fold lower than wild-type value
D258A
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turnover number is 522fold lower than wild-type value
E234A
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turnover number is 1.3fold higher than wild-type value
K162A
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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
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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
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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
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site-directed mutagenesis, 70fold increased Km for NADP+ compared to the wild-type enzyme
W252A
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site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
W252F
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site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
W252H
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site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
W252I
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site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
W252S
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site-directed mutagenesis, the mutant is no longer protected by Mn2+ against denaturation by urea and digestion by trypsin
Y91F
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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
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dimeric mutant enzyme with reduced activity compared to the wild type enzyme
D568A
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dimeric or tetrameric mutant enzyme with increased activity compared to the wild type enzyme
D90A
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dimeric or tetrameric mutant enzyme with reduced activity compared to the wild type enzyme
E314A
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site-directed mutagenesis
E314A/S346I/K347D/K362H
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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 inhibition by ATP compared to the wild-type enzyme
E314A/S346K
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site-directed mutagenesis
E314A/S346K/K347Y/K362H
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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 inhibition by ATP compared to the wild-type enzyme
E314A/S346K/K347Y/K362Q
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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 inhibition by ATP compared to the wild-type enzyme
H51A
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dimeric or tetrameric mutant enzyme with wild type activity
K347Y
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site-directed mutagenesis, the mutant enzyme shows a 5fold increased Km for NADP+ compared to the wild-type enzyme
K347Y/K362Q
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site-directed mutagenesis
K362H
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site-directed mutagenesis
K362Q
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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
K57S/E59N/K73E/D102S
site-directed mutagenesis, the mutant is primarily monomeric with some dimer formation
S346I/K347D/K362H
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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
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site-directed mutagenesis, site-directed mutagenesis, the mutant enzyme shows a 3fold increased Km for NADP+ compared to the wild-type enzyme
S346K/K347Y
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site-directed mutagenesis, the double mutant enzyme shows a 30fold increased Km for NADP+ compared to the wild-type enzyme
S346K/K347Y/K362H
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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
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site-directed mutagenesis, the triple c-NADP-ME mutant does not show significant reduction in its Km,NAD values
S346K/K362Q
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site-directed mutagenesis
S57K/N59E/E73K/S102D/H74K/D78P/D80E/D87G
S57K/N59E/E73K/S102D/H74K/D78P/D80E/D87G/D90E/K106S/Q121S/L125H
S57K/N59E/E73K/S102D/H74K/D78P/D80E/D87G/K106S/Q121S/L125H
moe
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enzyme inactivation by shRNA, ME1 activity is reduced by 62% and glucose-induced insulin secretion is decreased
I310V
site-directed mutagenesis
K228R
site-directed mutagenesis
R221G
site-directed mutagenesis
R221G/K228R
site-directed mutagenesis, substitution of Arg221 with Gly is responsible for the shift in reaction specificity
R221G/K228R/I310V
site-directed mutagenesis, the reaction specificity of the triple mutant is significantly shifted to malate production and the mutant gives a reduced amount of the byproduct than the wild-type. When the triple mutant enzyme is used as a catalyst for pyruvate carboxylation with NADH, the enzyme gives 1.2times higher concentration of malate than the wild-type with NADPH. Single-point mutation analysis reveals that the substitution of Arg221 with Gly is responsible for the shift in reaction specificity
V393V
1179 GTC /GTT results in a synonymous mutation of V393V
A339E
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the mutant of isoform nonC4-NADP-ME shows increased catalytic efficiency compared to the wild type enzyme
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
DelN
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the mutant of isoform nonC4-NADP-ME shows reduced catalytic efficiency compared to the wild type enzyme and is not inhibited by (S)-malate
DelN/A339E
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the mutant of isoform nonC4-NADP-ME shows reduced catalytic efficiency compared to the wild type enzyme
DelN/I140F/A339E
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the mutant of isoform nonC4-NADP-ME shows increased catalytic efficiency compared to the wild type enzyme and is not inhibited by (S)-malate
E339A
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the mutant of isoform C4-NADP-ME shows reduced catalytic efficiency compared to the wild type enzyme and is not inhibited by (S)-malate
F140I
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the mutant of isoform C4-NADP-ME shows reduced catalytic efficiency compared to the wild type enzyme
I140F
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the mutant of isoform nonC4-NADP-ME shows reduced catalytic efficiency compared to the wild type enzyme and is not inhibited by (S)-malate
I140F/A339E
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the mutant of isoform nonC4-NADP-ME shows reduced catalytic efficiency compared to the wild type enzyme and is not inhibited by (S)-malate
K225I
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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
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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
L544F
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the mutant of isoform C4-NADP-ME shows reduced catalytic efficiency compared to the wild type enzyme
Q503E
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the mutant of isoform C4-NADP-ME shows reduced catalytic efficiency compared to the wild type enzyme
S419A
the variant presents 72.3% of the wild type activity
S419E
the variant presents 8.7% of the wild type activity. The mutation dramatically decreases the affinity for the cofactor, as saturation is not observed even at NADP+ concentrations as high as 5 mM
R115A
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site-directed mutagenesis, mutation of isozyme NADP-ME2, the mutant shows similar kinetics as the wild-type isozyme NADP-ME2, but loses the activation ability of fumarate
R115A
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site-directed mutagenesis of isozyme NADP-ME2, the mutant displays a marked inhibition in the presence of all the organic acids tested, also fumarate
E73K
site-directed mutagenesis
E73K
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
H142A
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dimeric mutant enzyme with increased activity compared to the wild type enzyme
H142A
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site-directed mutagenesis, a dimeric tetramer interface mutant
H142A/D568A
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dimeric mutant enzyme with reduced activity compared to the wild type enzyme
H142A/D568A
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site-directed mutagenesis, a dimeric tetramer interface mutant. The mutant dimer completely dissociates into monomers after a 2.5 M urea treatment
H51A/D139A
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dimeric or tetrameric mutant enzyme with reduced activity compared to the wild type enzyme
H51A/D139A
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site-directed mutagenesis, a dimeric dimer interface mutant
H51A/D90A
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dimeric or tetrameric mutant enzyme with reduced activity compared to the wild type enzyme
H51A/D90A
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site-directed mutagenesis, a dimeric dimer interface mutant. The mutant dimer completely dissociates into monomers after a 1.5 M urea treatment
N59E
site-directed mutagenesis
N59E
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
N59E/E73K
site-directed mutagenesis
N59E/E73K
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
N59E/E73K/S102D
site-directed mutagenesis
N59E/E73K/S102D
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
S102D
site-directed mutagenesis
S102D
the mutant shows decreased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
S57K
site-directed mutagenesis
S57K
the mutant shows increased turnover numbers for (S)-malate and NADP+ compared to the wild type enzyme
S57K/N59E/E73K
site-directed mutagenesis
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
site-directed mutagenesis, the mutant is tetrameric
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
site-directed mutagenesis, the mutant is tetrameric
S57K/N59E/E73K/S102D/H74K/D78P/D80E/D87G/D90E/K106S/Q121S/L125H
site-directed mutagenesis
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
site-directed mutagenesis
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
W572A
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dimeric mutant enzyme with slightly reduced activity compared to the wild type enzyme
W572A
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site-directed mutagenesis, a dimeric tetramer interface mutant
C192A
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
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
C231A
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
C246A
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
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
C270A
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
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
R237L
site directed mutagenesis, mutation at the NADP+ binding site, mutant shows 530fold decreased kcat and 36.3 and 15.3fold increased Km for NADP+ and L-malate, respectively, compared to the wild-type enzyme, no activity with NAD+
R237L
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site-directed mutagenesis, mutation of a conserved residue involved in catalysis and substrate binding, mutant shows and an over 100fold higher partitioning ratio of oxaloacetate and malate compared to the wild-type enzyme, preference for reduction of oxaloacetate instead of decarboxylation
additional information
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construction of deletion mutants of isozyme NADP-ME2
additional information
expression of NADP-MDH is not affected in knock-out plants, knock-out mutant lines At5g58340::tDNA-1, SALK_053119, and At5g58340::tDNA-2, SALK_018118, carrying a DNA insert in the 5' region
additional information
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generation of knock-out mutants nadp-me2.1 and -2.2 by Agrobacterium tumefaciens strain GV3101-mediated transformation using the vacuum infiltration method
additional information
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recombinant enzyme overexpression in Chlorella pyrenoidosa for enhanced lipid production, method, overview
additional information
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effects of enzyme overexpression on the concentration of CO2 and the C4 metabolsim in wild-type strain and strains overexpressing other anaeplerotic enzymes, phenotypes, overview
additional information
deletion of gene maeB by chromosomal homologous recombination
additional information
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deletion of gene maeB by chromosomal homologous recombination
additional information
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deletion of gene maeB by chromosomal homologous recombination
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additional information
AF288898
construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei, expression pattern and regulation, overview
additional information
AF288899
construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei, expression pattern and regulation, overview
additional information
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construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei, expression pattern and regulation, overview
additional information
AF288898
construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei, expression patterns and regulation, overview
additional information
AF288899
construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei, expression patterns and regulation, overview
additional information
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construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei, expression patterns and regulation, overview
additional information
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generation of an antisense construct targeting the C4 isoform of NADP-malic enzyme, transformation of Flaveria bidentis via Agrobacterium tumefaciens, transgenic Flaveria bidentis plants exhibit a 34% to 75% reduction in NADP-ME activity relative to the wild-type with no visible growth phenotype. In transgenic plants, CO2 assimilation rates at high intercellular CO2 are significantly reduced, whereas the in vitro activities of both phosphoenolpyruvate carboxylase and Rubisco are increased
additional information
AF288911
construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs bearing parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei
additional information
construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs bearing parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei
additional information
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construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs bearing parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei
additional information
AF288911
construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei
additional information
construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei
additional information
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construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei
additional information
construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei
additional information
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construction of transgenic seedlings of Flaveria bidentis, expressing heterologous constructs beasring parts of isozymes ChlME1 and ChlME2 from Flaveria trinervia and Flaveria pringlei
additional information
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ME1 overexpression results in an increased glucose-dependent rise in malate and citrate levels in INS-1 832/13 cells. Introduction of ME1 activity alters glucose-stimulated insulin secretion to a lesser degree in mouse islets than in INS-1 832/13 cells, overview. In contrast to rat, mouse beta-cells lack ME1 activity, which is suggested to explain their lack of methyl succinate-stimulated insulin secretion. Metabolic phenotypes of transfected cells, overview
additional information
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a series of E314A-containing c-NADP-ME quadruple mutants are changed to NAD+-utilizing enzymes by abrogating NADP+ binding and increasing NAD+ binding. Abolishing the repulsive effect of Glu314 in the quadruple mutants increases the binding affinity of NAD+
additional information
multiple residues corresponding to the fumarate-binding site are mutated in human c-NADP-ME to correspond to those found in human m-NAD(P)-ME, EC 1.1.1.39. No significant difference between the wild-type and mutant enzymes in Km values for NADP+ and malate, and in kcat values. A chimeric enzyme, [51-105]_c-NADP-ME, is designed to include the putative fumarate-binding site ofm-NAD(P)-ME at the dimer interface of c-NADP-ME, but the chimera remains nonallosteric
additional information
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multiple residues corresponding to the fumarate-binding site are mutated in human c-NADP-ME to correspond to those found in human m-NAD(P)-ME, EC 1.1.1.39. No significant difference between the wild-type and mutant enzymes in Km values for NADP+ and malate, and in kcat values. A chimeric enzyme, [51-105]_c-NADP-ME, is designed to include the putative fumarate-binding site ofm-NAD(P)-ME at the dimer interface of c-NADP-ME, but the chimera remains nonallosteric
additional information
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expression of the enzyme from Mucor circinelloides as a strategy to improve lipid content inside the Rhodotorula glutinis yeast cells, overview. Heterologous expression of NADP+-dependent ME involved in fatty acid biosynthesis increases the lipid accumulation in the oleaginous yeast Rhodotorula glutinis
additional information
expression of the enzyme from Mucor circinelloides as a strategy to improve lipid content inside the Rhodotorula glutinis yeast cells, overview. Heterologous expression of NADP+-dependent ME involved in fatty acid biosynthesis increases the lipid accumulation in the oleaginous yeast Rhodotorula glutinis
additional information
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expression of malic enzyme from Mucor circinelloides as a strategy to improve lipid content inside the cells. The 26S rDNA and 5.8S rDNA gene fragments, with PGK1 gene from Saccharomyces cerevisiae strain 2.1445, isolated from Rhodotorula glutinis strain GM4 are used for homologous integration of the malc enzyme gene into Rhodotorula glutinis chromosome under the control of the constitutively highly expressed gene phosphoglycerate kinase 1 resulting in stable expression. Improvment of the lipid content by more than twofold. There are no significant differences in fatty acid profiles between the wild-type strain and the recombinant strain of Rhodotorula glutinis
additional information
expression of malic enzyme from Mucor circinelloides as a strategy to improve lipid content inside the cells. The 26S rDNA and 5.8S rDNA gene fragments, with PGK1 gene from Saccharomyces cerevisiae strain 2.1445, isolated from Rhodotorula glutinis strain GM4 are used for homologous integration of the malc enzyme gene into Rhodotorula glutinis chromosome under the control of the constitutively highly expressed gene phosphoglycerate kinase 1 resulting in stable expression. Improvment of the lipid content by more than twofold. There are no significant differences in fatty acid profiles between the wild-type strain and the recombinant strain of Rhodotorula glutinis
additional information
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expression of the enzyme from Mucor circinelloides as a strategy to improve lipid content inside the Rhodotorula glutinis yeast cells, overview. Heterologous expression of NADP+-dependent ME involved in fatty acid biosynthesis increases the lipid accumulation in the oleaginous yeast Rhodotorula glutinis
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additional information
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expression of malic enzyme from Mucor circinelloides as a strategy to improve lipid content inside the cells. The 26S rDNA and 5.8S rDNA gene fragments, with PGK1 gene from Saccharomyces cerevisiae strain 2.1445, isolated from Rhodotorula glutinis strain GM4 are used for homologous integration of the malc enzyme gene into Rhodotorula glutinis chromosome under the control of the constitutively highly expressed gene phosphoglycerate kinase 1 resulting in stable expression. Improvment of the lipid content by more than twofold. There are no significant differences in fatty acid profiles between the wild-type strain and the recombinant strain of Rhodotorula glutinis
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additional information
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construction of transgenic tobacco plants by expression of the maize enzyme, recombinant expression leads to plants with altered malate metabolism in guard cells and stomatal function which are more drought tolerant than the wild-type tobacco, overview
additional information
overexpression of rice isozyme NADP-ME2 in Arabidopsis thaliana enhances tolerance to salt and osmotic stresses in transgenic seedlings, overview
additional information
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overexpression of rice isozyme NADP-ME2 in Arabidopsis thaliana enhances tolerance to salt and osmotic stresses in transgenic seedlings, overview
additional information
transgenic Arabidopsis thaliana plants overexpressing rice cytosolic NADP-ME have a greater salt tolerance at the seedling stage than wild-type plants in MS medium-supplemented with different levels of NaCl, no obvious morphological or developmental differences between the transgenic and wild-type plants
additional information
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transgenic Arabidopsis thaliana plants overexpressing rice cytosolic NADP-ME have a greater salt tolerance at the seedling stage than wild-type plants in MS medium-supplemented with different levels of NaCl, no obvious morphological or developmental differences between the transgenic and wild-type plants
additional information
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siRNA knockdown of cytosolic ME1 by 89% of mRNA expression and enzyme activity significantly reduces glucose and decreases the flux of both pools of pyruvate through pyruvate carboxylase affecting the anaplerotic pathways, insulin secretion in response to membrane depolarization using potassium chloride is unaffected by siRNA knockdown of malic enzyme, overview
additional information
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siRNA knockout of ME1 in INS-1 832/13 beta-cells, siRNA knockdown and isotopic labeling strategies, method optimization, overview
additional information
generation of several ME3 knockdown cell lines, knockdown of inhibits insulin release. In the Me3 double-knockdown cells, the level of Me1 mRNA is not significantly decreased in the Me3-628(P)/Me2-725(H) and Me3-1672(P)/Me2-725(H) cell lines. However, a 32%, 49%, and 47% decrease in ME1 activity is observed in the cell lines Me3-628(P), Me3-628(P)/Me2-725(H), and Me3-628(P)/Me2-2124(H), respectively. Me3 mRNA is decreased in the Me3 single-knockdown cell line Me3-628(P), and it is lowered further in the Me3 and Me1 double-knockdown Me3628(P)/Me1-753(H) but not Me3-1672(P)/Me1-753(H). The level of Me3 mRNA in Me3-628(P)/Me1-753(H) and Me3-1672(P)/Me1-753(H) is 10% and 59% of the CHS control, respectively
additional information
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the tme gene is placed under the control of the dme promoter, and despite elevated levels of TME within bacteroids, no symbiotic nitrogen fixation occurred in dme mutant strains, the dme mutant cells fail to fix N2 in alfalfa root nodules, overview, no TME in the tme mutant RmG994
additional information
directed evolution of the thermotolerant NADP(H)-dependent malic enzyme from Thermococcus kodakarensis is conducted to alter the cofactor preference of the enzyme from NADP(H) to NAD(H). Integration of the thermotolerant NADPH-dependent malic enzyme (EC 1.1.1.40) from Thermococcus kodakarensis (TkME) to the chimeric EM pathway enables the construction of a cofactor-balanced and HCO3- fixing synthetic pathway, through which the direct conversion of glucose to malate can be achieved. The thermal degradation of the redox cofactors NADP+ and NADPH tends to be a major obstacle to the long-term operation of the in vitro metabolic system because, unlike living biological systems, it is not equipped with the complete enzyme apparatus for the de novo synthesis of these cofactors. No significant change is observed in the thermal stability of the wild type and mutant enzymes
additional information
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directed evolution of the thermotolerant NADP(H)-dependent malic enzyme from Thermococcus kodakarensis is conducted to alter the cofactor preference of the enzyme from NADP(H) to NAD(H). Integration of the thermotolerant NADPH-dependent malic enzyme (EC 1.1.1.40) from Thermococcus kodakarensis (TkME) to the chimeric EM pathway enables the construction of a cofactor-balanced and HCO3- fixing synthetic pathway, through which the direct conversion of glucose to malate can be achieved. The thermal degradation of the redox cofactors NADP+ and NADPH tends to be a major obstacle to the long-term operation of the in vitro metabolic system because, unlike living biological systems, it is not equipped with the complete enzyme apparatus for the de novo synthesis of these cofactors. No significant change is observed in the thermal stability of the wild type and mutant enzymes
additional information
construction of the Gsite5V and Gsite2V mutants, mutation at the NADP+ binding site
additional information
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construction of the Gsite5V and Gsite2V mutants, mutation at the NADP+ binding site
additional information
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construction of transgenic tobacco plants by expression of the maize enzyme, recombinant expression leads to plants with altered malate metabolism in guard cells and stomatal function which are more drought tolerant than the wild-type tobacco
additional information
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construction of chimerical NADP-ME sequences obtained from C4 and non-C4 isozymes
additional information
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transgenic Arabidopsis thaliana plants overexpressing the maize C4 NADP-malic enzyme show 6-33fold increase enzyme activity and a more rapid progression of dark-induced senescence compared to the wild type plants, overview