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0.011 - 207
ferricytochrome c
0.29 - 16.5
Phenyllactate
additional information
additional information
-
0.51
(S)-lactate
mutant Y254L with R289 distal, pH 7.0, 30°C
3.9
(S)-lactate
mutant D282N with R289 distal, pH 7.0, 30°C
13.2
(S)-lactate
-
pH 7.0, 25°C, mutant Y254F
14.18
(S)-lactate
-
pH 7.5, 25°C
270
(S)-lactate
wild-type enzyme with R289 distal, pH 7.0, 30°C
372
(S)-lactate
-
pH 7.0, 25°C, wild type
6
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant A67Q
39
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
62
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
63
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant L65A
87
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant P44A
108
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant A67L
139
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant E63K
143
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant P64R
155
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, wild-type enzyme
164
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant D72A
165
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant K73A
168
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant P64Q
171
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant V70M
184
cytochrome c
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant N69K
185
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant A67Q
190
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant L65A
196
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
207
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant P64Q
208
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant A67L
209
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
211
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant P44A
212
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant P64R
214
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant D72A
214
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, wild-type enzyme
218
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant E63K
218
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant N69K
223
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant K73A
225
FAD
pH 7.0, 30°C, flavin turnover in pre-steady-state flavin reduction, mutant V70M
0.011
ferricytochrome c
-
pH 7.5, 25°C, Tris-HCl buffer, mutant Y143F
0.02
ferricytochrome c
-
pH 7.5, 25°C, recombinantly expressed flavocytochrome b2 flavin-binding domain
0.103
ferricytochrome c
-
pH 7.5, 25°C, Tris-HCl buffer, wild-type enzyme
20
ferricytochrome c
-
pH 7.0, 5°C, phosphate buffer, mutant Y143F
61
ferricytochrome c
-
pH 7.0, 30°C, phosphate buffer, mutant Y143F
61
ferricytochrome c
-
pH 7.0, 5°C, phosphate buffer, wild-type enzyme
155
ferricytochrome c
-
pH 7.0, 30°C, phosphate buffer, wild-type enzyme
207
ferricytochrome c
-
pH 7.5, 25°C, wild-type enzyme
0.2
Fluoropyruvate
-
Y254F mutant enzyme
2.8
Fluoropyruvate
-
wild type enzyme
0.031
L-lactate
mutant H373Q, intact protein
0.057
L-lactate
mutant H373Q, using flavin domain only
0.39
L-lactate
-
Y254L mutant enzyme
0.42
L-lactate
-
Y254L mutant
6.1
L-lactate
-
Y254F mutant enzyme
7.5
L-lactate
-
Y254F mutant
7.8
L-lactate
-
influence of anions (400 mM KBr) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 2 mM ferricyanide and variable L-lactate concentration
8.6
L-lactate
-
mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 2 mM ferricyanide and variable L-lactate concentration
8.8
L-lactate
-
influence of anions (400 mM potassium acetate) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 2 mM ferricyanide and variable L-lactate concentration
9.2
L-lactate
-
influence of anions (200 mM phsophate) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 2 mM ferricyanide and variable L-lactate concentration
9.2
L-lactate
-
influence of anions (400 mM KCl) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 2 mM ferricyanide and variable L-lactate concentration
11
L-lactate
-
Y143F mutant with cytochrome c as electron acceptor
30
L-lactate
-
L230A mutant enzyme
41
L-lactate
-
A198G/L230A mutant enzyme
41
L-lactate
-
A198G/L230G mutant enzyme with ferricyanide as electron acceptor
45
L-lactate
-
influence of anions (400 mM KBr) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration
60
L-lactate
-
Y143F mutant enzyme
60
L-lactate
-
influence of anions (400 mM KCl) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration
61
L-lactate
-
influence of anions (400 mM KBr) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration
71
L-lactate
-
influence of anions (200 mM phsophate) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration
75
L-lactate
-
influence of anions (400 mM KCl) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration
86
L-lactate
-
influence of anions (400 mM potassium acetate) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration
101
L-lactate
-
influence of anions (300 mM phsophate) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration
103
L-lactate
-
native enzyme with cytochrome c as electron acceptor
113
L-lactate
-
influence of anions (400 mM potassium acetate) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration
117
L-lactate
-
holo-enzyme, reaction conditions: 1 or 2 mM ferricyanide, 100 mM phosphate buffer, 1mM EDTA, pH 7, 5°C
133
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C
144
L-lactate
-
stopped-flow kinetic parameters for flavin reduction by L-lactate using holo-enzyme, reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, in the absence of ferrocyanide
149
L-lactate
-
stopped-flow kinetic parameters for flavin reduction by L-lactate using FDH domain, reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, in the absence of ferrocyanide
155
L-lactate
-
wild type enzyme
185
L-lactate
-
A198G mutant enzyme
200
L-lactate
wild-type, using flavin domain only
214
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 10 mM Tris/HCl buffer, 0.1 M NaCl, pH 7.5, 25°C
240
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7.5, 25°C
259
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 30°C
270
L-lactate
-
wild type enzyme
372
L-lactate
wild-type, intact protein
400
L-lactate
-
wild type enzyme
400
L-lactate
-
native enzyme and Y143F mutant with ferricyanide as electron acceptor
473
L-lactate
-
wild type enzyme
0.02
L-Mandelate
-
wild type enzyme with ferricyanide as electron acceptor
8.5
L-Mandelate
-
A198G/L230G mutant enzyme with ferricyanide as electron acceptor
0.29
Phenyllactate
-
Y254F mutant enzyme
16.5
Phenyllactate
-
wild type enzyme
additional information
additional information
-
systematic determination of substrates with different chain length
-
additional information
additional information
-
systematic determination of activity of all mutants with L-mandelate and L-lactate as substrate
-
additional information
additional information
-
systematic determination of turnover numbers for wild type enzyme and deletion mutants with different electron acceptors, significantly reduced activity for deletion mutants with cytochrome c as electron acceptor
-
additional information
additional information
-
second-order rate constant for cytochrome c reduction in the pre-steady-state determined by stopped-flow spectrophotometry
-
additional information
additional information
-
steady-state kinetic parameters and 2H kinetic isotope effects for the isolated flavin domain and intact, wild-type flavocytoochrome b2
-