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2'-dATP + riboflavin
2'-dADP + riboflavin 5'-phosphate
-
-
-
-
?
ADP + riboflavin
AMP + FMN
-
22% of the activity with ATP
-
-
?
ATP + 10-(D-allo)flavin
ADP + 10-(D-allo)flavin 5'-phosphate
-
30% of the activity with riboflavin
-
-
?
ATP + 10-(L-arabo)flavin
ADP + 10-(L-arabo)flavin 5'-phosphate
-
25% of the activity with riboflavin
-
-
?
ATP + 2'-deoxyriboflavin
ADP + 2'-deoxyriboflavin 5'-phosphate
-
31% of the activity with riboflavin
-
-
?
ATP + 2-thioriboflavin
ADP + 2-thioriboflavin 5'-phosphate
ATP + 3-deazariboflavin
ADP + 3-deazariboflavin 5'-phosphate
-
-
-
-
?
ATP + 3-methylriboflavin
ADP + 3-methylriboflavin 5'-phosphate
-
5% of the activity with riboflavin
-
-
?
ATP + 5-deazariboflavin
ADP + 5-deazariboflavin 5'-phosphate
ATP + 5-deazariboflavin
ADP + 5-deazariboflavin monophosphate
-
-
-
?
ATP + 5-methyl-5-deazariboflavin
ADP + 5-methyl-5-deazariboflavin 5'-phosphate
-
-
-
-
?
ATP + 6,7-dichloro-9-(D-1'-ribityl)isoalloxazine
ADP + 6,7-dichloro-9-(D-1'-ribityl)isoalloxazine phosphate
ATP + 6,7-dimethyl-9-(1'-D-ribityl)-2-iminoisoalloxazine
ADP + 6,7-dimethyl-9-(1'-D-ribityl)-2-iminoisoalloxazine 5'-phosphate
-
18% of the activity with riboflavin
-
-
?
ATP + 6-methylriboflavin
ADP + 6-methylriboflavin 5'-phosphate
-
-
-
-
?
ATP + 7,8-dichlororiboflavin
ADP + 7,8-dichlororiboflavin monophosphate
-
-
-
?
ATP + 7-chlororiboflavin
ADP + 7-chlororiboflavin
-
-
-
-
?
ATP + 8-aminoriboflavin
ADP + 8-aminoriboflavin monophosphate
-
-
-
?
ATP + 8-bromo-8-demethylriboflavin
ADP + 8-bromo-8-demethylriboflavin 5-'phosphate
-
384% of the activity with riboflavin
-
-
?
ATP + 8-chloro-8-demethylriboflavin
ADP + 8-chloro-8-demethylriboflavin 5'-phosphate
-
122.2% of the activity with riboflavin
-
-
?
ATP + 8-demethylriboflavin
ADP + 8-demethylriboflavin 5'-phosphate
ATP + 8-dimethylamino-8-demethylriboflavin
ADP + 8-dimethylamino-8-demethylriboflavin 5'-phosphate
ATP + 8-ethoxy-8-demethylriboflavin
ADP + 8-ethoxy-8-demethylriboflavin 5'-phosphate
-
210% of the activity with riboflavin
-
-
?
ATP + 8-fluoro-8-demethylriboflavin
ADP + 8-fluoro-8-demethylriboflavin 5'-phosphate
-
132.2% of the activity with riboflavin
-
-
?
ATP + 8-iodo-8-demethylriboflavin
ADP + 8-iodo-8-demethylriboflavin 5'-phosphate
-
334.7% of the activity with riboflavin
-
-
?
ATP + 8-methoxy-8-demethylriboflavin
ADP + 8-methoxy-8-demethylriboflavin 5'-phosphate
-
114.5% of the activity with riboflavin
-
-
?
ATP + 8-methylamino-8-demethylriboflavin
ADP + 8-methylamino-8-demethylriboflavin 5'-phosphate
-
237.3% of the activity with riboflavin
-
-
?
ATP + 9-azariboflavin
ADP + 9-azariboflavin 5'-phosphate
-
-
-
-
?
ATP + alloflavin
ADP + alloflavin 5'-phosphate
-
-
-
-
?
ATP + arabitylflavin
ADP + arabitylflavin phosphate
ATP + D-erythroflavin
ADP + D-erythroflavin 5'-phosphate
ATP + riboflavin
ADP + FMN
ATP + roseoflavin
ADP + roseoflavin 5'-phosphate
CTP + riboflavin
CDP + riboflavin 5'-phosphate
-
50% of the activity with ATP
-
-
?
dATP + riboflavin
dADP + FMN
GTP + riboflavin
GDP + riboflavin 5'-phosphate
-
no activity
-
-
?
riboflavin + ATP
FMN + ADP
-
-
-
-
?
UTP + riboflavin
UDP + FMN
-
31% of the activity with ATP
-
-
?
additional information
?
-
ATP + 2-thioriboflavin
ADP + 2-thioriboflavin 5'-phosphate
-
60% of the activity with riboflavin
-
-
?
ATP + 2-thioriboflavin
ADP + 2-thioriboflavin 5'-phosphate
-
30% of the activity with riboflavin
-
-
?
ATP + 5-deazariboflavin
ADP + 5-deazariboflavin 5'-phosphate
-
-
-
-
?
ATP + 5-deazariboflavin
ADP + 5-deazariboflavin 5'-phosphate
-
-
-
-
?
ATP + 5-deazariboflavin
ADP + 5-deazariboflavin 5'-phosphate
-
-
-
-
?
ATP + 5-deazariboflavin
ADP + 5-deazariboflavin 5'-phosphate
-
15% of the activity with riboflavin
-
-
?
ATP + 5-deazariboflavin
ADP + 5-deazariboflavin 5'-phosphate
-
as active as riboflavin
-
-
?
ATP + 6,7-dichloro-9-(D-1'-ribityl)isoalloxazine
ADP + 6,7-dichloro-9-(D-1'-ribityl)isoalloxazine phosphate
-
52% of the activity with riboflavin
-
-
?
ATP + 6,7-dichloro-9-(D-1'-ribityl)isoalloxazine
ADP + 6,7-dichloro-9-(D-1'-ribityl)isoalloxazine phosphate
-
35% of the activity with riboflavin
-
-
?
ATP + 8-demethylriboflavin
ADP + 8-demethylriboflavin 5'-phosphate
-
35% of the activity with riboflavin
-
-
?
ATP + 8-demethylriboflavin
ADP + 8-demethylriboflavin 5'-phosphate
-
110% of the activity with riboflavin
-
-
?
ATP + 8-dimethylamino-8-demethylriboflavin
ADP + 8-dimethylamino-8-demethylriboflavin 5'-phosphate
-
110% of the activity with riboflavin
-
-
?
ATP + 8-dimethylamino-8-demethylriboflavin
ADP + 8-dimethylamino-8-demethylriboflavin 5'-phosphate
-
70% of the activity with riboflavin
-
-
?
ATP + arabitylflavin
ADP + arabitylflavin phosphate
-
14% of the activity with riboflavin
-
-
?
ATP + arabitylflavin
ADP + arabitylflavin phosphate
-
slightly more active than riboflavin
-
-
?
ATP + D-erythroflavin
ADP + D-erythroflavin 5'-phosphate
-
-
-
-
?
ATP + D-erythroflavin
ADP + D-erythroflavin 5'-phosphate
-
10-(D-erythro)flavin, 33% of the activity with riboflavin
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
?
ATP + riboflavin
ADP + FMN
-
specific for the reduced form of riboflavin
-
-
?
ATP + riboflavin
ADP + FMN
-
key enzyme in flavin biosynthesis
-
-
?
ATP + riboflavin
ADP + FMN
ribC is essential for growth of Bacillus subtilis. RibC is not directly involved in the riboflavin regulatory system
-
-
?
ATP + riboflavin
ADP + FMN
-
the flavokinase activity appears to be localized to the N-terminal domain of the protein. RibR specifically interacts in vivo with the nontranslated wild-type leader of the mRNA of the riboflavin biosynthetic operon. In RibR itself, interaction was localized to the carboxy-terminate part of the protein
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
ribC is essential for growth of Bacillus subtilis. RibC is not directly involved in the riboflavin regulatory system
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
ir
ATP + riboflavin
ADP + FMN
essential enzyme catalyzing the phosphorylation of riboflavin to form FMN, an obligatory step in vitamin B2 utilization and flavin cofactor synthesis
-
-
?
ATP + riboflavin
ADP + FMN
substrate and product binding structure, overview
-
-
ir
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
key enzyme in flavin biosynthesis
-
-
?
ATP + riboflavin
ADP + FMN
-
ATP in form of MgATP2-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
-
?
ATP + riboflavin
ADP + FMN
-
-
-
?
ATP + roseoflavin
ADP + roseoflavin 5'-phosphate
-
-
-
-
?
ATP + roseoflavin
ADP + roseoflavin 5'-phosphate
-
187% of the activity with 8-demethylriboflavin
-
-
?
ATP + roseoflavin
ADP + roseoflavin 5'-phosphate
-
81% of the activity with riboflavin
-
-
?
ATP + roseoflavin
ADP + roseoflavin 5'-phosphate
-
90% of the activity with riboflavin
-
-
?
ATP + roseoflavin
ADP + roseoflavin 5'-phosphate
-
-
-
?
dATP + riboflavin
dADP + FMN
-
-
-
-
?
dATP + riboflavin
dADP + FMN
-
-
-
-
?
additional information
?
-
ribC wild-type gene product has both flavokinase and flavin adenine dinucleotide synthetase activity
-
-
?
additional information
?
-
-
ribC wild-type gene product has both flavokinase and flavin adenine dinucleotide synthetase activity
-
-
?
additional information
?
-
-
the riboflavin kinase encoding gene ribR of Bacillus subtilis is a part of a 10 kb operon, which is negatively regulated by the yrzC gene product
-
-
?
additional information
?
-
-
ribC wild-type gene product has both flavokinase and flavin adenine dinucleotide synthetase activity
-
-
?
additional information
?
-
bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26
-
-
?
additional information
?
-
-
bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26
-
-
?
additional information
?
-
bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26
-
-
?
additional information
?
-
-
bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26
-
-
?
additional information
?
-
bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26
-
-
?
additional information
?
-
-
bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26
-
-
?
additional information
?
-
the engineered FAD synthetase from Corynebacterium ammoniagenes with deleted N-terminal adenylation domain is a biocatalyst that is stable and efficient for direct and quantitative phosphorylation of riboflavin and riboflavin analogues to their corresponding FMN cofactors at preparative-scale, method evaluation, overview
-
-
?
additional information
?
-
-
the engineered FAD synthetase from Corynebacterium ammoniagenes with deleted N-terminal adenylation domain is a biocatalyst that is stable and efficient for direct and quantitative phosphorylation of riboflavin and riboflavin analogues to their corresponding FMN cofactors at preparative-scale, method evaluation, overview
-
-
?
additional information
?
-
riboflavin kinase activity of the FAD synthetase (EC 2.7.7.2) from Corynebacterium ammoniagenes. Adenine and flavin nucleotide ligands cooperate in their binding to the RFK module
-
-
-
additional information
?
-
-
riboflavin kinase activity of the FAD synthetase (EC 2.7.7.2) from Corynebacterium ammoniagenes. Adenine and flavin nucleotide ligands cooperate in their binding to the RFK module
-
-
-
additional information
?
-
substrate binding structure analysis using crystal structures of substrate-bound RFK. Binding of HsRFK substrates is the kinetically preferred process compared to product inhibition, kinetic modeling, overview
-
-
-
additional information
?
-
-
substrate binding structure analysis using crystal structures of substrate-bound RFK. Binding of HsRFK substrates is the kinetically preferred process compared to product inhibition, kinetic modeling, overview
-
-
-
additional information
?
-
-
no phosphorylation of isoriboflavin, galactoflavin, dulcitylflavin, sorbitylflavin
-
-
?
additional information
?
-
the bifunctional flavokinase/flavin adenine dinucleotide synthetase produces inactive flavin cofactors and is not involved in resistance to the antibiotic roseoflavin
-
-
?
additional information
?
-
-
the bifunctional flavokinase/flavin adenine dinucleotide synthetase produces inactive flavin cofactors and is not involved in resistance to the antibiotic roseoflavin
-
-
?
additional information
?
-
bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26
-
-
?
additional information
?
-
the flavin motif is involved in flavin ligand binding, role of active site residues in the catalytic mechanism, overview. The isoalloxazine ring is sandwiched between the indole ring of Trp184 and the planar guanidinium group of Arg189, while the hydrophilic pyrimidine ring forms two specific hydrogen bonds between its C4 carbonyl and the main chain amide of Asp181, and between its N3 amide and the side chain of Asp181, respectively. Residues Asn62, Asp66, Asp168, and Arg297 interact either with ATP phosphate groups, or to coordinate the catalytic Mg2+ ion either directly or indirectly through water molecules. Arg297 might be involved in the interaction with the phosphate groups of both substrates, and helps in their positioning for the nucleophilic attack in the adenylyltransfer reaction
-
-
?
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0.0000103 - 0.18
riboflavin
additional information
additional information
-
0.000021
ATP
-
pH 8.5, 30°C
0.000021
ATP
-
ATP in form of MgATP2-
0.0002
ATP
-
bifunctional enzyme AtFMN/FHy
0.005
ATP
-
ATP in form of MgATP2-
0.0083
ATP
-
pH 7.0, 25°C
0.01
ATP
mutant enzyme S164D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.011
ATP
mutant enzyme R161D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.011
ATP
mutant enzyme T165D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.012
ATP
mutant enzyme H28D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.012
ATP
mutant enzyme R161A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.012
ATP
mutant enzyme S164A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.012
ATP
mutant enzyme T165A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0137
ATP
-
apparent value, wild type enzyme, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.014
ATP
mutant enzyme H28A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.014
ATP
mutant enzyme H31A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.014
ATP
wild type enzyme, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0176
ATP
-
apparent value, mutant enzyme E268D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.0215
ATP
mutant enzyme R66E, at pH 7.0 and 25°C
0.0215
ATP
pH 7.0, 25°C, recombinant mutant R66E
0.024
ATP
mutant enzyme N125A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0282
ATP
pH 7.0, 25°C, recombinant wild-type enzyme
0.0282
ATP
wild type enzyme, at pH 7.0 and 25°C
0.03
ATP
pH 7.0, 25°C, recombinant enzyme
0.035
ATP
mutant enzyme N125D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.04
ATP
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Lineweaver-Burk equation kinetics
0.0453
ATP
-
apparent value, mutant enzyme N210D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.06
ATP
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Michaelis-Menten kinetics
0.087
ATP
pH 7.0, 25°C, recombinant mutant R66A
0.0000103
riboflavin
-
bifunctional enzyme AtFMN/FHy
0.00012
riboflavin
-
pH 8.5, 30°C
0.00032
riboflavin
-
allosteric kinetics
0.00089
riboflavin
mutant enzyme R66E, at pH 7.0 and 25°C
0.00089
riboflavin
pH 7.0, 25°C, recombinant mutant R66E
0.0017
riboflavin
mutant enzyme N125A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0021
riboflavin
-
apparent value, mutant enzyme N210D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.0025
riboflavin
pH 7.0, 25°C, recombinant enzyme
0.0041
riboflavin
-
apparent value, mutant enzyme E268D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.0045
riboflavin
mutant enzyme H31A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0047
riboflavin
-
pH 7.2, 37°C
0.0056
riboflavin
mutant enzyme S164D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0069
riboflavin
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Lineweaver-Burk equation kinetics
0.0077
riboflavin
mutant enzyme T165D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0088
riboflavin
mutant enzyme S164A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.01
riboflavin
-
pH 8.0, 37°C
0.01
riboflavin
-
pH 7.2, 37°C
0.01
riboflavin
mutant enzyme T165A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.01
riboflavin
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Michaelis-Menten kinetics
0.0117
riboflavin
pH 7.0, 25°C, recombinant wild-type enzyme
0.0117
riboflavin
wild type enzyme, at pH 7.0 and 25°C
0.012
riboflavin
mutant enzyme R161A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.013
riboflavin
-
apparent value, wild type enzyme, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.013
riboflavin
mutant enzyme R161D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.013
riboflavin
wild type enzyme, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0153
riboflavin
-
pH 7.0, 25°C
0.016
riboflavin
mutant enzyme N125D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.023
riboflavin
mutant enzyme H28A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.025
riboflavin
mutant enzyme H28D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.18
riboflavin
-
pH 8, 37°C
0.03
roseoflavin
-
37°C
additional information
additional information
steady-state kinetic analysis of wild-type and mutant enzymes
-
additional information
additional information
bi-substrate enzyme kinetics of the recombinant DELTA(1-182)CaFADS module
-
additional information
additional information
-
bi-substrate enzyme kinetics of the recombinant DELTA(1-182)CaFADS module
-
additional information
additional information
Michaelis-Menten model and thermodynamic profiles of recombinant wild-type and mutant enzymes, overview
-
additional information
additional information
-
Michaelis-Menten model and thermodynamic profiles of recombinant wild-type and mutant enzymes, overview
-
additional information
additional information
Michaelis-Menten kinetics and modeling, cooperativity model, pre-steady-state kinetics/pre-steady-state stopped-flow kinetics and dissociation constants, overview. Isothermal titration calorimetry, and Gibbs free energy flow for the interaction of HsRFK with substrates and products. Thermodynamics modulates the ligand binding landscape of HsRFK. Cooperativity coefficients, ANP and FLV ligands cooperate in their binding to HsRFK
-
additional information
additional information
-
Michaelis-Menten kinetics and modeling, cooperativity model, pre-steady-state kinetics/pre-steady-state stopped-flow kinetics and dissociation constants, overview. Isothermal titration calorimetry, and Gibbs free energy flow for the interaction of HsRFK with substrates and products. Thermodynamics modulates the ligand binding landscape of HsRFK. Cooperativity coefficients, ANP and FLV ligands cooperate in their binding to HsRFK
-
additional information
additional information
Michaelis-Menten steady-state kinetic study. While Km ATP values increase with the ADP concentration, kcat values remain constant. The high affinity for the ADP product inhibitor considerably increases the estimated error for KmATP. Association and dissociation kinetics of flavin ligands to the RFK module are measured by flavin fluorescence changes, stopped-flow kinetics. Pre-steady-state kinetic analysis of the binding of flavins to the RFK module of CaFADS, thermodynamic diagram for the RFK-ligand interactions. Adenine and flavin nucleotide ligands cooperate in their binding to the RFK module. Detailed kinetic analysis, overview
-
additional information
additional information
-
Michaelis-Menten steady-state kinetic study. While Km ATP values increase with the ADP concentration, kcat values remain constant. The high affinity for the ADP product inhibitor considerably increases the estimated error for KmATP. Association and dissociation kinetics of flavin ligands to the RFK module are measured by flavin fluorescence changes, stopped-flow kinetics. Pre-steady-state kinetic analysis of the binding of flavins to the RFK module of CaFADS, thermodynamic diagram for the RFK-ligand interactions. Adenine and flavin nucleotide ligands cooperate in their binding to the RFK module. Detailed kinetic analysis, overview
-
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0.042
ATP
-
apparent value, mutant enzyme N210D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.07
ATP
-
apparent value, mutant enzyme E268D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.4
ATP
mutant enzyme R66E, at pH 7.0 and 25°C
0.5
ATP
pH 7.0, 25°C, recombinant mutant R66A
0.7
ATP
pH 7.0, 25°C, recombinant mutant R66E
0.75
ATP
mutant enzyme H28A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.75
ATP
mutant enzyme T165D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.77
ATP
mutant enzyme N125A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.92
ATP
mutant enzyme H28D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.92
ATP
mutant enzyme S164D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.97
ATP
mutant enzyme H31A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.98
ATP
mutant enzyme S164A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
1.08
ATP
mutant enzyme R161A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
1.1
ATP
mutant enzyme T165A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
1.13
ATP
-
apparent value, wild type enzyme, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
1.13
ATP
mutant enzyme R161D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
1.13
ATP
wild type enzyme, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
1.33
ATP
mutant enzyme N125D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
1.7
ATP
pH 7.0, 25°C, recombinant enzyme
2.17
ATP
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Lineweaver-Burk equation kinetics
2.6
ATP
wild type enzyme, at pH 7.0 and 25°C
2.67
ATP
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Michaelis-Menten kinetics
6.8
ATP
pH 7.0, 25°C, recombinant wild-type enzyme
0.045
riboflavin
-
apparent value, mutant enzyme E268D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.085
riboflavin
-
apparent value, mutant enzyme N210D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
0.1
riboflavin
-
pH 8.5, 30°C
0.5
riboflavin
pH 7.0, 25°C, recombinant mutant R66A
0.7
riboflavin
mutant enzyme R66E, at pH 7.0 and 25°C
0.7
riboflavin
pH 7.0, 25°C, recombinant mutant R66E
1.7
riboflavin
pH 7.0, 25°C, recombinant enzyme
1.85
riboflavin
mutant enzyme N125A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
2.82
riboflavin
mutant enzyme H31A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
2.85
riboflavin
mutant enzyme T165A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
3
riboflavin
mutant enzyme T165D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
3.08
riboflavin
mutant enzyme S164D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
4.32
riboflavin
mutant enzyme S164A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
4.78
riboflavin
mutant enzyme H28D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
5
riboflavin
-
apparent value, wild type enzyme, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
5
riboflavin
mutant enzyme R161A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
5
riboflavin
mutant enzyme R161D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
5.02
riboflavin
wild type enzyme, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
5.33
riboflavin
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Lineweaver-Burk equation kinetics
6.8
riboflavin
pH 7.0, 25°C, recombinant wild-type enzyme
6.8
riboflavin
wild type enzyme, at pH 7.0 and 25°C
6.92
riboflavin
mutant enzyme N125D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
7.12
riboflavin
mutant enzyme H28A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
7.33
riboflavin
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Michaelis-Menten kinetics
0.4
roseoflavin
-
37°C
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0.83
ATP
-
apparent value, mutant enzyme N210D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
3.83
ATP
-
apparent value, mutant enzyme E268D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
18.3
ATP
mutant enzyme R66E, at pH 7.0 and 25°C
31.7
ATP
mutant enzyme N125A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
38.3
ATP
mutant enzyme N125D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
44.5
ATP
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Michaelis-Menten kinetics
51.7
ATP
mutant enzyme H28A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
54.25
ATP
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Lineweaver-Burk equation kinetics
56.67
ATP
pH 7.0, 25°C, recombinant enzyme
68.3
ATP
mutant enzyme T165D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
71.7
ATP
mutant enzyme H31A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
78.3
ATP
mutant enzyme H28D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
81.7
ATP
wild type enzyme, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
82.17
ATP
-
apparent value, wild type enzyme, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
83.3
ATP
mutant enzyme S164A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
88.3
ATP
mutant enzyme S164D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
90
ATP
mutant enzyme T165A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
91.6
ATP
pH 7.0, 25°C, recombinant wild-type enzyme
93.3
ATP
mutant enzyme R161A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
101.7
ATP
mutant enzyme R161D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
582
ATP
wild type enzyme, at pH 7.0 and 25°C
788.3
ATP
pH 7.0, 25°C, recombinant mutant R66E
0.0758
riboflavin
pH 7.0, 25°C, recombinant mutant R66E
0.142
riboflavin
pH 7.0, 25°C, recombinant mutant R66A
10.83
riboflavin
-
apparent value, mutant enzyme E268D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
40.3
riboflavin
-
apparent value, mutant enzyme N210D, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
91.7
riboflavin
wild type enzyme, at pH 7.0 and 25°C
200
riboflavin
mutant enzyme H28D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
283
riboflavin
mutant enzyme T165A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
300
riboflavin
mutant enzyme H28A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
383
riboflavin
mutant enzyme R161D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
383
riboflavin
mutant enzyme T165D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
383
riboflavin
wild type enzyme, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
386
riboflavin
-
apparent value, wild type enzyme, at 37°C in 20 mM PIPES pH 7.0, 0.8 mM MgCl2
417
riboflavin
mutant enzyme R161A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
433
riboflavin
mutant enzyme N125D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
483
riboflavin
mutant enzyme S164A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
550
riboflavin
mutant enzyme S164D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
581.7
riboflavin
pH 7.0, 25°C, recombinant wild-type enzyme
617
riboflavin
mutant enzyme H31A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
680
riboflavin
pH 7.0, 25°C, recombinant enzyme
733
riboflavin
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Michaelis-Menten kinetics
772.5
riboflavin
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Lineweaver-Burk equation kinetics
788
riboflavin
mutant enzyme R66E, at pH 7.0 and 25°C
1083
riboflavin
mutant enzyme N125A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
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0.0065
1'-DL-glyceryl-6,7-dimethylisoalloxazine
-
competitive
0.021
1-Deazariboflavin
-
pH 8, 37°C
0.008
10-(5'-Hydroxypentyl)flavin
-
pH 8, 37°C
0.007
10-(Hydroxyethyl)flavin
-
pH 8, 37°C
0.0071
3'-hydroxypropyl-6,7-dimethylisoalloxazine
-
competitive
0.41
3-Deazariboflavin
-
pH 8, 37°C
0.0076
4'-hydroxybutyl-6,7-dimethylisoalloxazine
-
competitive
0.0078
5'-hydroxypentyl-6,7-dimethylisoalloxazine
-
competitive
0.275
5-Deazariboflavin
-
pH 8, 37°C
0.25
8-Aminoriboflavin
-
pH 8, 37°C
0.02
8-Diethylaminoriboflavin
-
pH 8, 37°C
0.016
8-Ethoxyriboflavin
-
pH 8, 37°C
0.175
8-Ethylaminoriboflavin
-
pH 8, 37°C
0.5
8-hydroxyriboflavin
-
pH 8, 37°C
0.015
8-Methoxyriboflavin
-
pH 8, 37°C
0.47
8-Methylaminoriboflavin
-
pH 8, 37°C
0.03
8-Methylethylaminoriboflavin
-
pH 8, 37°C
0.0079
9-(6'-hydroxyhexyl)-6,7-dimethylisoalloxazine
-
competitive
2.67
ATP
pH 7.0, 25°C, recombinant RFK activity of the RFK module
0.01
lumichrome
-
pH 8, 37°C
0.007
Lumiflavin
-
pH 8, 37°C
0.0018 - 0.0063
riboflavin
0.018
riboflavin 5'-phosphate
-
pH 8, 37°C
additional information
additional information
-
0.017
ADP
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Lineweaver-Burk equation kinetics
0.033
ADP
pH 7.0, 25°C, recombinant enzyme
0.0014
FMN
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Lineweaver-Burk equation kinetics
0.0025
FMN
pH 7.0, 25°C, recombinant enzyme
0.006
FMN
-
pH 8.0, 37°C, against riboflavin
0.0018
riboflavin
mutant enzyme H28A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0019
riboflavin
mutant enzyme H28D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0019
riboflavin
pH 7.0, 25°C, recombinant RFK activity of the RFK module, Michaelis-Menten kinetics
0.0032
riboflavin
mutant enzyme T165A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0033
riboflavin
mutant enzyme N125D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.004
riboflavin
wild type enzyme, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0041
riboflavin
mutant enzyme T165D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0044
riboflavin
mutant enzyme S164A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0046
riboflavin
mutant enzyme N125A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0051
riboflavin
mutant enzyme R161A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0051
riboflavin
mutant enzyme R161D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0058
riboflavin
mutant enzyme S164D, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.0063
riboflavin
mutant enzyme H31A, in 20 mM PIPES, 0.8 mM MgCl2, pH 7.0, at 37°C
0.023
ZnADP-
-
pH 8.0, 37°C, against ZnATP2-
0.12
ZnADP-
-
pH 8.0, 37°C, against riboflavin
additional information
additional information
inhibition kinetics, recombinant enzyme, overview
-
additional information
additional information
-
inhibition kinetics, recombinant enzyme, overview
-
additional information
additional information
inhibition kinetics, dead-end inhibition by substrate excess
-
additional information
additional information
-
inhibition kinetics, dead-end inhibition by substrate excess
-
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malfunction
enzyme RFK downregulation alters expression profiles of clock-controlled metabolic-genes and destroys flavins protection on stroke treatments, while its activity reduction links to protein-energy malnutrition and thyroid hormones decrease
metabolism
biosynthesis of FMN and FAD from riboflavin (RF) involves two reactions: RF is first phosphorylated to FMN in an ATP-Mg2+-dependent reaction carried out by an ATP:riboflavin kinase (RFK), and then an FMN:ATP adenylyltransferase (FMNAT) transfers the adenylyl group from a second ATP molecule to FMN to yield FAD. In eukaryotes, these reactions are preferentially performed by two independent monofunctional enzymes, but in most prokaryotes, the two reactions are sequentially catalyzed by a bifunctional enzyme known as prokaryotic type I FAD synthetase (FADS). These bifunctional proteins are organized in two nearly independent modules with each one catalyzing one of the two activities
evolution
eukaryotic FMNAT is related to phosphoadenosine phosphosulfate (PAPS) reductase family proteins and contains a core domain with a modified Rossman-fold topology and a C-terminal extension
evolution
whereas the N-terminal module of FADS lacks structural homology to eukaryotic FMNATs, the kinase module is homologous to monofunctional RFKs
evolution
different organisms, different regulatory strategies of RFKs, overview
physiological function
-
riboflavin kinase is a TNF-receptor-1 binding protein that couples TNF-receptor-1 to NADPH oxidase
physiological function
-
riboflavin kinase is a TNF-receptor-1 binding protein that couples TNF-receptor-1 to NADPH oxidase
physiological function
-
the enzyme plays a critical role in the KD548-Fc-mediated reactive oxygen species accumulation and downstream signaling. The enzyme is essential in recruiting Nox1 to death receptor4/5
physiological function
flavocoenzymes, including flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), are versatile redox cofactors involved in many fundamental cellular processes in all living organisms. FAD is synthesized from riboflavin obtained from the diet via two enzymatic steps catalyzed by riboflavin kinase (RFK, EC 2.7.1.26) and essential FMN adenylyltransferase (FMNAT,EC 2.7.7.2). Phosphorylation of riboflavin by RFK is crucial for specific absorption of the vitamin and is the physiologically rate-limiting step in the biosynthesis of flavocoenzymes, whereas product (FAD) feedback inhibition is observed for mammalian FMNAT, suggesting that biosynthesis of FAD is also regulated at the FMNAT reaction step
physiological function
the essential cofactors of flavoproteins and flavoenzymes, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), are synthesized from riboflavin in two sequential reactions: riboflavin phosphorylation is catalysed by an ATP-riboflavin kinase (RFK, EC 2.7.1.26) to produce FMN, which can be then converted to FAD by an FMN:ATP adenylyltranferase (FMNAT, EC 2.7.7.2). Bacteria contain a single bifunctional polypeptide called FAD synthetase (FADS)
physiological function
Enzymes known as bifunctional and bimodular prokaryotic type-I FAD synthetase (FADS) exhibit ATP:riboflavin kinase (RFK) and FMN:ATP adenylyltransferase (FMNAT) activities in their C-terminal and N-terminal modules, respectively, and produce flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD). These act as cofactors of a plethora of flavoproteins in all organisms. Kinetics and thermodynamics of the protein-ligand interactions of riboflavin kinase activity of FAD synthetase from Corynebacterium ammoniagenes, overview. FMN synthesis is a key process that requires tight regulation. FMN production by CaFADS is highly regulated by substrate and product inhibition of its RFK activity to avoid overproduction even though the RF substrate might transiently increases in the media, where usually the amount of RF is considerably lower than that of FMN and particularly of FAD (either free or as part of flavoproteins)
physiological function
human riboflavin kinase is an essential enzyme that catalyzes the biosynthesis of the flavin mononucleotide (FMN) cofactor using riboflavin (RF, vitamin B2) and ATP as substrates. Human riboflavin kinase (HsRFK) catalyzes vitamin B2 (riboflavin) phosphorylation to flavin mononucleotide (FMN), obligatory step in flavin cofactor synthesis. HsRFK expression is related to protection from oxidative stress, amyloid-beta toxicity, and some malignant cancers progression. HsRFK is also predicted as involved in a protein-protein association network that at the system level affects to different cellular processes
additional information
molecular dynamics simulations of riboflavin kinase domain bound to FMN, ADP, and Mg2+, structure-function analysis, flavin-binding site structure in the RFK module of CaFADS, overview
additional information
-
molecular dynamics simulations of riboflavin kinase domain bound to FMN, ADP, and Mg2+, structure-function analysis, flavin-binding site structure in the RFK module of CaFADS, overview
additional information
residue E268 is the catalytic base of the kinase reaction. The salt bridge between E268 at the RFK site and R66 at the FMNAT-module is important for the riboflavinkinase activity. Cross-talk between the RFK- and FMNAT-modules of neighboring protomers in the CaFADS enzyme, and participation of R66 in the modulation of the geometry of the RFK active site during catalysis
additional information
-
residue E268 is the catalytic base of the kinase reaction. The salt bridge between E268 at the RFK site and R66 at the FMNAT-module is important for the riboflavinkinase activity. Cross-talk between the RFK- and FMNAT-modules of neighboring protomers in the CaFADS enzyme, and participation of R66 in the modulation of the geometry of the RFK active site during catalysis
additional information
adenine and flavin nucleotide ligands cooperate in their binding to the RFK module
additional information
-
adenine and flavin nucleotide ligands cooperate in their binding to the RFK module
additional information
molecular dynamics simulations, structure-function analysis
additional information
-
molecular dynamics simulations, structure-function analysis
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E268D
-
the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme
H28A
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
H28D
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
H31A
the mutant shows increased catalytic efficiency compared to the wild type enzyme
N125A
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
N125D
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
N210D
-
the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme
R161A
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
R161D
the mutant shows wild type catalytic efficiency
R66X
point mutations at R66 have only mild effects on ligand binding and kinetic properties of the FMNAT-module (where R66 is located), but considerably impair the RFK activity turnover. Substitutions of R66 also modulate the ratio between monomeric and oligomeric species and modify the quaternary arrangement observed by single-molecule methods
S164A
the mutant shows increased catalytic efficiency compared to the wild type enzyme
S164D
the mutant shows increased catalytic efficiency compared to the wild type enzyme
T165A
the mutant shows reduced catalytic efficiency compared to the wild type enzyme
T165D
the mutant shows wild type catalytic efficiency
E86Q
-
destroying the kinase domain, purified as C-terminal glutathione S-transferase fusionprotein
N36D
-
purified as C-terminal glutathione S-transferase fusionprotein
D168A
site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme
D181A
site-directed mutagenesis, the mutant shows reduced sensitivity to inhibition by FAD compared to the wild-type enzyme and has a much faster turnover rate than the wild-type enzyme
D66A
site-directed mutagenesis, inactive mutant
N62A
site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme
N62S
site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme
R297A
site-directed mutagenesis, the mutant shows a 2fold increased activity compared to the wild-type enzyme
R297A/R300A
site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme
R300A
site-directed mutagenesis, the mutant shows 93% reduced activity compared to the wild-type enzyme
W184A
site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme
R66A
inactive
R66A
site-directed mutagenesis, R66A CaFADS shows a considerable increase in the amount of oligomeric species
R66E
the mutant shows increased activity compared to the wild type enzyme
R66E
site-directed mutagenesis, R66E CaFADS shows a considerable increase in the amount of oligomeric species
additional information
engineering of the FAD synthetase from Corynebacterium ammoniagenes by deleting its N-terminal adenylation domain leads to a biocatalyst that is stable and efficient for direct and quantitative phosphorylation of riboflavin and riboflavin analogues to their corresponding FMN cofactors at preparative-scale. Deletion of the N-terminal adenosyl transfer domain in the truncated C-terminal RF kinase domain, tcRFK, variants results in a drop in the TM value from 40°C (parental CaFADS) to 35°C for tcRFK. Addition of the C-terminal poly-His tag further reduces the TM to 30°C, presumably due to the conformationally flexible tail formed by the extra amino acids
additional information
-
engineering of the FAD synthetase from Corynebacterium ammoniagenes by deleting its N-terminal adenylation domain leads to a biocatalyst that is stable and efficient for direct and quantitative phosphorylation of riboflavin and riboflavin analogues to their corresponding FMN cofactors at preparative-scale. Deletion of the N-terminal adenosyl transfer domain in the truncated C-terminal RF kinase domain, tcRFK, variants results in a drop in the TM value from 40°C (parental CaFADS) to 35°C for tcRFK. Addition of the C-terminal poly-His tag further reduces the TM to 30°C, presumably due to the conformationally flexible tail formed by the extra amino acids
additional information
isolation of the RFK module of CaFADS (DELTA(1-182)CaFADS). This truncated form of the enzyme has shown to perform the RFK activity with ligand binding profiles and strong substrate inhibition that are similar to those observed in the full-length bifunctional enzyme
additional information
-
isolation of the RFK module of CaFADS (DELTA(1-182)CaFADS). This truncated form of the enzyme has shown to perform the RFK activity with ligand binding profiles and strong substrate inhibition that are similar to those observed in the full-length bifunctional enzyme
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