BRENDA - Enzyme Database show
show all sequences of 1.1.1.248

Removal of substrate inhibition and increase in maximal velocity in the short chain dehydrogenase/reductase salutaridine reductase involved in morphine biosynthesis

Ziegler, J.; Brandt, W.; Geissler, R.; Facchini, P.J.; J. Biol. Chem. 284, 26758-26767 (2009)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
overexpression in Escherichia coli SG13009
Papaver somniferum
Engineering
Amino acid exchange
Commentary
Organism
D107A
increase in Km, 2fold decrease in kcat
Papaver somniferum
F104A
increase in kcat
Papaver somniferum
F104A/I275A
shows no substrate inhibition accompanied by a weak affinity for productive salutaridine binding, kcat is almost 2fold higher compared with the wild-type
Papaver somniferum
I275A
increase in Km, shows an increase of velocity by a factor of 2.3
Papaver somniferum
K186V
moderate effects on substrate affinity, exhibits a turnover number similar to that of the wild-type
Papaver somniferum
L185A
increase in Km, decrease in kcat
Papaver somniferum
L266A
decrease in kcat
Papaver somniferum
M271A
decrease in kcat; increase in Km
Papaver somniferum
N272A
increase in Km, decrease in kcat
Papaver somniferum
S181A
moderate effects on substrate affinity, increase in kcat by 50%
Papaver somniferum
T182A
moderate effects on substrate affinity
Papaver somniferum
V106A
moderate effects on substrate affinity, increase in kcat by 90%
Papaver somniferum
Inhibitors
Inhibitors
Commentary
Organism
Structure
additional information
strong substrate inhibition of SalR
Papaver somniferum
NADP+
product inhibition
Papaver somniferum
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0021
-
salutaridine
wild-type
Papaver somniferum
0.0034
-
NADPH
mutant K186V
Papaver somniferum
0.0035
-
NADPH
wild-type
Papaver somniferum
0.0041
-
salutaridine
mutant T182A
Papaver somniferum
0.0045
-
NADPH
mutant S181A
Papaver somniferum
0.005
-
salutaridine
mutant S181A
Papaver somniferum
0.007
-
NADPH
mutant V106A
Papaver somniferum
0.0078
-
salutaridine
mutant K186V
Papaver somniferum
0.0089
-
salutaridine
mutant V106A
Papaver somniferum
0.0097
-
NADPH
mutant F104A
Papaver somniferum
0.0132
-
salutaridine
mutant L185A
Papaver somniferum
0.0148
-
salutaridine
mutant D107A
Papaver somniferum
0.0155
-
NADPH
mutant L266A
Papaver somniferum
0.0156
-
NADPH
mutant N272A
Papaver somniferum
0.0198
-
NADPH
mutant M271A
Papaver somniferum
0.0211
-
NADPH
mutant I275A
Papaver somniferum
0.0213
-
NADPH
mutant T182A
Papaver somniferum
0.0225
-
salutaridine
mutant M271A
Papaver somniferum
0.0297
-
NADPH
mutant L185A
Papaver somniferum
0.0305
-
NADPH
mutant D107A
Papaver somniferum
0.0318
-
salutaridine
mutant F104A
Papaver somniferum
0.0331
-
salutaridine
mutant I275A
Papaver somniferum
0.043
-
NADPH
mutant F104A/I275A
Papaver somniferum
0.0494
-
salutaridine
mutant L266A
Papaver somniferum
0.0901
-
salutaridine
mutant N272A
Papaver somniferum
0.4086
-
salutaridine
mutant F104A/I275A
Papaver somniferum
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Papaver somniferum
Q071N0
-
-
Purification (Commentary)
Commentary
Organism
to homogeneity
Papaver somniferum
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
salutaridine + NADPH + H+
-
698937
Papaver somniferum
(7S)-salutaridinol + NADP+
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.006
-
salutaridine
mutant M271A
Papaver somniferum
0.016
-
salutaridine
mutant L185A
Papaver somniferum
0.021
-
salutaridine
mutant N272A
Papaver somniferum
0.126
-
salutaridine
mutant T182A
Papaver somniferum
0.501
-
salutaridine
mutant L266A
Papaver somniferum
0.61
-
salutaridine
mutant D107A
Papaver somniferum
0.998
-
salutaridine
mutant K186V
Papaver somniferum
1.306
-
salutaridine
wild-type
Papaver somniferum
1.933
-
salutaridine
mutant S181A
Papaver somniferum
2.45
-
salutaridine
mutant F104A/I275A
Papaver somniferum
2.458
-
salutaridine
mutant V106A
Papaver somniferum
2.727
-
salutaridine
mutant F104A
Papaver somniferum
3.188
-
salutaridine
mutant I275A
Papaver somniferum
Cofactor
Cofactor
Commentary
Organism
Structure
NADPH
-
Papaver somniferum
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.0012
-
NADP+
-
Papaver somniferum
Cloned(Commentary) (protein specific)
Commentary
Organism
overexpression in Escherichia coli SG13009
Papaver somniferum
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
-
Papaver somniferum
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D107A
increase in Km, 2fold decrease in kcat
Papaver somniferum
F104A
increase in kcat
Papaver somniferum
F104A/I275A
shows no substrate inhibition accompanied by a weak affinity for productive salutaridine binding, kcat is almost 2fold higher compared with the wild-type
Papaver somniferum
I275A
increase in Km, shows an increase of velocity by a factor of 2.3
Papaver somniferum
K186V
moderate effects on substrate affinity, exhibits a turnover number similar to that of the wild-type
Papaver somniferum
L185A
increase in Km, decrease in kcat
Papaver somniferum
L266A
decrease in kcat
Papaver somniferum
M271A
decrease in kcat; increase in Km
Papaver somniferum
N272A
increase in Km, decrease in kcat
Papaver somniferum
S181A
moderate effects on substrate affinity, increase in kcat by 50%
Papaver somniferum
T182A
moderate effects on substrate affinity
Papaver somniferum
V106A
moderate effects on substrate affinity, increase in kcat by 90%
Papaver somniferum
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
additional information
strong substrate inhibition of SalR
Papaver somniferum
NADP+
product inhibition
Papaver somniferum
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.0012
-
NADP+
-
Papaver somniferum
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0021
-
salutaridine
wild-type
Papaver somniferum
0.0034
-
NADPH
mutant K186V
Papaver somniferum
0.0035
-
NADPH
wild-type
Papaver somniferum
0.0041
-
salutaridine
mutant T182A
Papaver somniferum
0.0045
-
NADPH
mutant S181A
Papaver somniferum
0.005
-
salutaridine
mutant S181A
Papaver somniferum
0.007
-
NADPH
mutant V106A
Papaver somniferum
0.0078
-
salutaridine
mutant K186V
Papaver somniferum
0.0089
-
salutaridine
mutant V106A
Papaver somniferum
0.0097
-
NADPH
mutant F104A
Papaver somniferum
0.0132
-
salutaridine
mutant L185A
Papaver somniferum
0.0148
-
salutaridine
mutant D107A
Papaver somniferum
0.0155
-
NADPH
mutant L266A
Papaver somniferum
0.0156
-
NADPH
mutant N272A
Papaver somniferum
0.0198
-
NADPH
mutant M271A
Papaver somniferum
0.0211
-
NADPH
mutant I275A
Papaver somniferum
0.0213
-
NADPH
mutant T182A
Papaver somniferum
0.0225
-
salutaridine
mutant M271A
Papaver somniferum
0.0297
-
NADPH
mutant L185A
Papaver somniferum
0.0305
-
NADPH
mutant D107A
Papaver somniferum
0.0318
-
salutaridine
mutant F104A
Papaver somniferum
0.0331
-
salutaridine
mutant I275A
Papaver somniferum
0.043
-
NADPH
mutant F104A/I275A
Papaver somniferum
0.0494
-
salutaridine
mutant L266A
Papaver somniferum
0.0901
-
salutaridine
mutant N272A
Papaver somniferum
0.4086
-
salutaridine
mutant F104A/I275A
Papaver somniferum
Purification (Commentary) (protein specific)
Commentary
Organism
to homogeneity
Papaver somniferum
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
salutaridine + NADPH + H+
-
698937
Papaver somniferum
(7S)-salutaridinol + NADP+
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.006
-
salutaridine
mutant M271A
Papaver somniferum
0.016
-
salutaridine
mutant L185A
Papaver somniferum
0.021
-
salutaridine
mutant N272A
Papaver somniferum
0.126
-
salutaridine
mutant T182A
Papaver somniferum
0.501
-
salutaridine
mutant L266A
Papaver somniferum
0.61
-
salutaridine
mutant D107A
Papaver somniferum
0.998
-
salutaridine
mutant K186V
Papaver somniferum
1.306
-
salutaridine
wild-type
Papaver somniferum
1.933
-
salutaridine
mutant S181A
Papaver somniferum
2.45
-
salutaridine
mutant F104A/I275A
Papaver somniferum
2.458
-
salutaridine
mutant V106A
Papaver somniferum
2.727
-
salutaridine
mutant F104A
Papaver somniferum
3.188
-
salutaridine
mutant I275A
Papaver somniferum
Other publictions for EC 1.1.1.248
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
720707
Wijekoon
Systematic knockdown of morphi ...
Papaver somniferum
Plant J.
69
1052-1063
2012
-
-
1
-
1
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
725460
Higashi
Atomic structure of salutaridi ...
Papaver somniferum
J. Biol. Chem.
286
6532-6541
2011
-
-
1
1
15
-
-
-
-
-
-
1
-
3
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
1
15
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
3
3
-
-
-
710745
Higashi
Crystallization and preliminar ...
Papaver somniferum
Acta Crystallogr. Sect. F
66
163-166
2010
-
-
1
1
-
-
-
-
-
-
-
-
-
4
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
698937
Ziegler
Removal of substrate inhibitio ...
Papaver somniferum
J. Biol. Chem.
284
26758-26767
2009
-
-
1
-
12
-
2
26
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
13
-
-
-
1
1
-
-
-
-
1
1
-
12
-
-
2
1
26
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
13
-
-
-
-
-
-
-
-
-
-
700676
Ziegler
Evolution of morphine biosynth ...
Papaver arenarium, Papaver bracteatum, Papaver pilosum, Papaver pyrenaicum, Papaver somniferum
Phytochemistry
70
1696-1707
2009
-
-
1
-
-
-
-
-
-
-
-
-
-
8
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
5
-
-
-
-
-
1
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
700683
Kempe
RNAi suppression of the morphi ...
Papaver somniferum
Phytochemistry
70
579-589
2009
-
-
1
-
-
-
1
-
1
-
-
-
-
3
-
-
1
-
-
1
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
1
-
-
1
-
-
-
-
-
-
1
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
689571
Geissler
Molecular modeling and site-di ...
Papaver bracteatum
Plant Physiol.
143
1493-1503
2007
-
-
1
1
13
-
1
6
-
-
2
2
-
4
-
-
1
-
-
-
-
-
4
1
1
-
-
5
2
1
-
4
1
1
-
-
-
1
4
1
13
-
-
1
1
6
-
-
2
2
-
-
-
1
-
-
-
-
4
1
1
-
-
5
2
1
-
1
-
-
-
-
-
-
670562
Ziegler
Comparative transcript and alk ...
Papaver somniferum, Papaver somniferum salR
Plant J.
48
177-192
2006
-
-
1
-
-
-
-
4
-
-
3
4
-
6
-
-
1
-
-
-
-
-
8
1
1
1
-
-
2
2
-
4
-
1
-
-
-
1
4
-
-
-
-
-
-
4
-
-
3
4
-
-
-
1
-
-
-
-
8
1
1
1
-
-
2
2
-
1
-
-
-
-
-
-
9554
Gerardy
-
Purification and characterizat ...
no activity in Papaver alpinum, no activity in Papaver atlanticum, no activity in Papaver dubium, no activity in Papaver feddei, no activity in Papaver lateritium, no activity in Papaver oreophilum, no activity in Papaver orientale, no activity in Papaver persicum, no activity in Papaver pilosum, no activity in Papaver rhoeas, no activity in Papaver rupifragum, no activity in Papaver strigosum, no activity in Papaver tauricula, no activity in Papaver triniifolium, Papaver bracteatum, Papaver somniferum
Phytochemistry
34
125-132
1993
-
-
-
-
-
-
-
4
1
-
1
1
-
16
-
-
1
1
-
6
1
1
2
1
1
-
-
-
2
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
1
-
1
1
-
-
-
1
-
6
1
1
2
1
1
-
-
-
2
2
1
-
-
-
-
-
-
-