Information on EC 1.1.1.30 - 3-hydroxybutyrate dehydrogenase

New: Word Map on EC 1.1.1.30
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
1.1.1.30
-
RECOMMENDED NAME
GeneOntology No.
3-hydroxybutyrate dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(R)-3-hydroxybutanoate + NAD+ = acetoacetate + NADH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Butanoate metabolism
-
-
ketogenesis
-
-
ketolysis
-
-
Metabolic pathways
-
-
Synthesis and degradation of ketone bodies
-
-
SYSTEMATIC NAME
IUBMB Comments
(R)-3-hydroxybutanoate:NAD+ oxidoreductase
Also oxidizes other 3-hydroxymonocarboxylic acids.
CAS REGISTRY NUMBER
COMMENTARY hide
9028-38-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Acidovorax sp.
strain SA1
SwissProt
Manually annotated by BRENDA team
strain T
-
-
Manually annotated by BRENDA team
strain T
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
enzyme shows hysteresis, lag phase in progress curve
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
Gln94, His144, Lys152, and Gln196 form hydrogen bonds with carboxyl and/or ketone group of acetoacetate, Trp187, Trp257 form hydrophobic interactions with the carbon atoms of acetoacetate, and Ser142 and Tyr155 are directly related to the catalytic mechanism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-2-hydroxybutanoate + NAD+
acetoacetate + NADH
show the reaction diagram
-
isoenzyme from heavy mitochondria and isoenzyme from light mitochondria show no significant difference in activity
-
-
?
(R)-3-hydroxybutanoate + 3-acetylpyridine adenine dinucleotide
acetoacetate + 3-acetylpyridine adenine dinucleotideH2
show the reaction diagram
-
-
-
r
(R)-3-hydroxybutanoate + NAD+
acetoacetate + NADH
show the reaction diagram
(R)-3-hydroxybutanoate + NAD+
acetoacetate + NADH + H+
show the reaction diagram
(R)-3-hydroxybutanoate + NADH + H+
acetoacetate + NAD+
show the reaction diagram
(R)-3-hydroxybutanoate + NADP+
acetoacetate + NADPH
show the reaction diagram
activity with NADP+ is 2% of the activity with NAD+
-
-
?
(R)-3-hydroxybutyrate + NAD+
acetoacetate + NADH + H+
show the reaction diagram
2-hydroxypropansulfonate + NAD+
acetonylsulfonate + NADH
show the reaction diagram
-
-
-
r
2-methyl-3-hydroxybutyrate + NAD+
2-methylacetoacetate + NADH
show the reaction diagram
-
-
-
r
4-hydroxybutanoate + NAD+
acetoacetate + NADH
show the reaction diagram
-
isoenzyme from heavy mitochondria shows 40% lower activity than enzyme fromlight mitochondria
-
-
?
D-3-hydroxybutyrate + NAD+
acetoacetate + NADH + H+
show the reaction diagram
L-threonine + NAD+
?
show the reaction diagram
-
-
-
?
poly-3-hydroxybutyrate + NAD+
acetoacetate + NADH + H+
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(R)-3-hydroxybutanoate + NAD+
acetoacetate + NADH
show the reaction diagram
(R)-3-hydroxybutanoate + NAD+
acetoacetate + NADH + H+
show the reaction diagram
-
-
-
-
r
(R)-3-hydroxybutanoate + NADH + H+
acetoacetate + NAD+
show the reaction diagram
-
the enzyme is involved in poly(3-hydroxybutyrate) biosynthesis together with acetoacetyl-CoA thiolase and PHB synthase, pathway overview, intracellular concentrations of key metabolites, i.e. CoA, acetyl-CoA, 3HB-CoA, NAD+/NADH, determine whether a cell accumulates or degrades PHB
-
-
r
(R)-3-hydroxybutyrate + NAD+
acetoacetate + NADH + H+
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADP+
-
2% of the activity with NAD+
additional information
-
no activity with NADP+/NADPH
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
10 mM, 2fold increase in activity
Cu2+
-
10 mM, 2fold increase in activity
Zn2+
-
10 mM, 2fold increase in activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(DL)-2-hydroxybutyrate
-
5 mM, pH 8.5, 25, 69% inhibition
(DL)-lactate
-
5 mM, pH 8.5, 25C, 65% inhibition
2,4-Dichlorophenoxyacetic acid
-
inhibits the enzyme, in vivo treatment of the animals with 2,4-dichlorophenoxyacetic acid leads to strong decrease of triglycerides level and HDL cholesterol and an increase in GOT level and LDL cholesterol, and necrosis of seminiferous tubules cells in testis, hyperplasia of hepatocytes in liver and presence of multinucleated giant cells in brain, overview
2-Hydroxybutyrate
-
10 mM, 57% inhibition
2-oxobutyrate
-
10 mM, 29% inhibition
3-hydroxybutyrate
-
substrate inhibition at concentrations above 20 mM, 50% inhibition at 200 mM, cyanylated enzyme is 3fold more active at high substrate concentration
4-mercapto-3-nitrobenzoate
-
derivatization of enzyme sulfhydryl groups, less than 1% residual activity
5,5'-dithiobis(2-nitrobenzoic acid)
-
0.15 mM, 63% inhibition
acetoacetate
acetoacetyl-CoA
-
1 mM, 29% inhibition
acetyl-CoA
-
1 mM, 82% inhibition
acetylphosphonate
-
16.7 mM, pH 7, 28% inhibition
ADP
-
2.85 mM, 0.025 mM NADH, 63% inhibition, competitive inhibitor vs. NADH and NAD+, pH 7-7.5, noncompetitive vs. acetoacetate
ADP-ribose
-
competitive inhibition vs. coenzyme, noncompetitive vs. substrate
AMP
-
2.85 mM, 0.025 mM NADH, 55% inhibition, competitive inhibitor vs. NADH and NAD+, pH 7-7.5
ATP
-
2.85 mM, 0.025 mM NADH, 79% inhibition, competitive inhibitor vs. NADH and NAD+, pH 7-7.5, noncompetitive vs. acetoacetate
bromomalonate
Butanedione
-
-
Butyrate
cacodylate
-
competitive
chloromalonate
crotonate
-
10 mM, 20% inhibition
CuSO4
-
1 mM, 81% inhibition
D-lactate
-
5 mM, pH 8.5, 25, 85% inhibition, uncompetitive vs. coenzyme, competitive vs. substrate
diazenedicarboxylic acid bis(dimethylamide)
dimethyl malonate
-
-
dimethyl phosphate
-
16.7 mM, pH 7, 12% inhibition
dimethylmalonate
dimethyloxyphosphinylacetate
-
competitive inhibitor vs. acetoacetate
DL-2-Hydroxybutyrate
-
-
DL-lactate
-
-
EDTA
Acidovorax sp.
inhibition and destabilization of the enzyme at 10 mM
glucose
Acidovorax sp.
inhibits the enzyme when contained in the growth medium
Hg2+
Acidovorax sp.
inhibition at 1 mM, and destabilization at 10 mM
Hydroxymalonate
KCl
-
80-100 mM, 50% inhibition
L-3-hydroxybutyrate
LiCl
-
80-100 mM, 50% inhibition
malonate
mesoxalate
methanephosphonic acid
-
3.3 mM, pH 7, 11% inhibition
methyl methylphosphonate
-
3.3 mM, pH 7, 13% inhibition
methyl phosphate
-
16.7 mM, pH 7, 21% inhibition
Methyl-2-methoxyphosphinylacetate
-
competitive inhibitor vs. acetoacetate
-
methylmalonate
MnCl2
-
1 mM, 50% inhibition
N-ethylmaleimide
NaCl
-
80-100 mM, 50% inhibition
NAD+
-
substrate inhibition, high concentrations, noncompetitive vs. beta-hydroxybutyrate
NH4Cl
-
80-100 mM, 50% inhibition
oxalate
-
16.7 mM, pH 7, 9% inhibition
p-chloromercuribenzoate
Phenylarsine oxide
-
NAD+ and NADH protect against inhibition
Phenylglyoxal
Propionate
-
5 mM, pH 8.5, 25C, 9% inhibition
pyruvate
sodium acetonylphosphonate
-
3.3 mM, pH 7, 21% inhibition
sodium monomethyl acetylphosphonate
-
16.7 mM, pH 7, 60% inhibition
sodium monomethylacetonylphosphate
-
16.7 mM, pH 7, 30% inhibition
Sodium sulfide
-
competitive vs. beta-hydroxybutyrate
sodium sulfite
-
uncompetitive vs. NAD+ and acetoacetate, competitive vs. beta-hydroxybutyrate
succinate
-
10 mM, 12% inhibition
Zn2+
Acidovorax sp.
inhibition and destabilization of the enzyme at 10 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
10 mM, 41% increase
3-hydroxybutyrate
Acidovorax sp.
induces the enzyme when contained in the growth medium
dithiothreitol
EDTA
-
0.5 mM, 21% increase
glycerol
Acidovorax sp.
-
KCN
-
0.1 mM, 37% increase
Lactate
Acidovorax sp.
induces the enzyme when contained in the growth medium
phosphatidylcholine
Phospholipids
-
absolutely dependent on
reduced glutathione
-
10 mM, 38% increase
succinate
Acidovorax sp.
induces the enzyme when contained in the growth medium
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.34 - 131
(R)-3-hydroxybutanoate
0.45 - 2
(R)-3-hydroxybutyrate
0.28
3-acetylpyridine adenine dinucleotide
-
-
0.00547 - 32
acetoacetate
0.0391 - 260
beta-hydroxybutyrate
0.34 - 100
D-3-hydroxybutyrate
388
L-threonine
-
-
0.000445 - 2.2
NAD+
0.00102 - 1.5
NADH
7.4
NADP+
-
-
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
13 - 1390
(R)-3-hydroxybutanoate
4.5 - 150
acetoacetate
6.1 - 705
D-3-hydroxybutyrate
87 - 360
NAD+
58 - 740
NADH
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.38 - 5.74
2,4-Dichlorophenoxyacetic acid
5.6
cacodylate
-
-
80
dimethyl malonate
-
-
0.81
DL-2-Hydroxybutyrate
-
-
0.9
DL-lactate
-
-
3.8
malonate
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.02
-
cofactor 3-acetylpyridine adenine dinucleotide
0.038
-
brain 3-hydroxybutyrate dehydrogenase, mitochondrial extracts
0.07
-
heart 3-hydroxybutyrate dehydrogenase, mitochondrial extracts; kidney 3-hydroxybutyrate dehydrogenase, mitochondrial extracts
0.09
-
purified enzyme
0.17
-
solubilized apo-3-hydroxybutyrate dehydrogenase, reconstituted with phosphatidylcholine/phosphatidylethanolamine/diphosphatidylglycerol, 5/4/1
0.22
-
apo-3-hydroxybutyrate dehydrogenase, reconstituted with phosphatidylcholine/phosphatidylethanolamine/diphosphatidylglycerol, 5/4/1
0.5
-
enzyme expressed in SF9 cells, S24T mutatant
0.52
-
submitochondrial vesicles, membrane bound enzyme
0.54
-
submitochondrial vesicles
0.6
-
enzyme expressed in SF9 cells, M92V mutatant
0.63
-
enzyme expressed in SF9 cells, membrane bound
0.8
-
enzyme expressed in SF9 cells, C242S mutatant
1.15
-
liver 3-hydroxybutyrate dehydrogenase, reconstituted with microsomal membranes
1.26
-
brain 3-hydroxybutyrate dehydrogenase, mitochondrial extracts
1.275
-
liver 3-hydroxybutyrate dehydrogenase, reconstituted with mitochondrial inner membranes
2.7
-
submitochondrial vesicles
23.8
-
-
41
-
purified enzyme, reconstituted with mitochondrial phospholipids
72
Acidovorax sp.
purified native enzyme
86
-
purified enzyme, reconstituted with mitochondrial phospholipids
91
-
liver 3-hydroxybutyrate dehydrogenase, reconstituted with mitochondrial phospholipids
100
-
-
123
-
-
145
-
heart 3-hydroxybutyrate dehydrogenase, reconstituted with mitochondrial phospholipids
300
Acidovorax sp.
purified recombinant enzyme
360
recombinant enzyme
450
recombinant enzyme
additional information
-
0.0018 mM/min/g wet weight
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 5
Acidovorax sp.
reduction reaction
6
-
reduction reaction
6 - 6.5
-
reduction of acetoacetate
6.4 - 7
-
reduction of acetoacetate
6.8
-
acetoacetate reduction
7.4
-
assay at
8.4
-
oxidation of beta-hydroxybutyrate
8.6
-
oxidation of (R)-3-hydroxybutanoate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 7.5
-
acetoacetate reduction
5.9 - 8.6
-
acetoacetate reduction, 50% activity at pH 5.0, 66% at pH 8.6
6.5 - 8.5
optimal pH of BDH3 is 8.5 in the oxidation reaction and 6.5 in the reduction reaction
7.6 - 9
-
oxidation of beta-hydroxybutyrate
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
55
Acidovorax sp.
-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 60
-
30% of maximal activity at 20C, 63% of maximal activity at 60C
25 - 49
-
80% activity at 25C and at 49C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
gastric glandular mucosa
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
68000
-
nondenaturing PAGE
89000
-
gel filtration
106000
-
submitochondrial vesicles, radiation inactivation
110000
112000
-
gel filtration
119000
-
reconstituted enzyme, radiation inactivation, irradiation leads to loss of activity and fragmentation
120000
-
gel filtration
132000
-
gel filtration
140000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystallization of the enzyme in the apo form and in the holo form with acetate as a substrate analogue, method screening, mother liquor consists of 30% w/v PEG 4000, 0.2 M sodium acetate trihydrate and 100 mM Tris-HCl, pH 8.5, at 20C, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular-replacement method
-
in the presence of the substrate D-3-hydroxybutyrate and the cofactor NAD+ at the optimum pH for the catalytic reaction. At 277 and 293 K using the hanging-drop vapour-diffusion method, to 2.3 A resolution. Structure is isomorphous to that of the complex with the substrate analogue acetate
-
sitting drop vapor diffusion method at 20C, structure determined at a resolution of 1.8 A in complex with NAD(H)
-
crystals of ternary complex of HBDH-NAD+-L-3-hydroxybutyrate and the binary complex of HBDH-NAD+. The former structure shows a closed-form conformation, which is considered an active form for catalysis, while the latter stays mostly in a open-form conformation. Crystals of mutants T190S and T190A
-
hanging-drop vapor-diffusion method, ligand-free enzyme and enzyme-NAD+ complex, 2.0 A resolution
-
hanging-drop vapour-diffusion method using PEG 3000 as a precipitating agent. The crystals belong to the orthorhombic group P2(1)2(1)2, with unit-cell parameters a = 64.3, b = 99.0, c = 110.2 A. The crystals are most likely to contain two tetrameric subunits in the asymmetric unit
hanging drop vapour diffusion method with 17-20% polyethylene glycol 1500, 0.1 M Tris-HCl, pH 7.1, 0.2 mM CaCl2 and 10 mM acetoacetate; recombinant enzyme, hanging drop vapor diffusion method, 0.003 ml of HBDH, 10 mg/ml, is mixed with an equal volume of crystallization buffer containing 17-20% PEG 1500, 0.1 M Tris-HCl, pH 7.1, 0.2 mM CaCl2, and 10 mM acetoacetate, room temperature/22C, three different crystal forms, X-ray diffraction structure determination and analysis at resolutions between 1.9 and 2.1 A
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0 - 40
Acidovorax sp.
-
0 - 37
-
stable at, 30% remaining activity after 40 min at 37C, oxidation reaction
0 - 45
-
stable at 0C and 25C for 90 min, 18% activity after 90 min at 45C, activity restored after 60 min at 37C, loss of activity at 60C
45
-
25% remaining activity after 20 min, oxidation reaction
60
-
28% remaining activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 1 mg enzyme/ml, 0.2 mM potassium phosphate pH 8.0, 10% glycerol, 10 mM 2-mercaptoethanol, 6 months, 0% inactivation, 0C, 0.1 mg enzyme/ml, 1 day, 50% inactivation
-
-20C, 20 mM phosphate buffer, pH 7.0, 20% glycerol
-20C, crude extract, partially purified enzyme, several weeks, 0% inactivation
-
-20C, several months, 0%, 4C, lyophilized, 2 month, 0% inactivation, 4C, 0.1 M sodium phosphate buffer pH 7-8, 5 days, 50% inactivation, stability increases in the presence of 1-10 mM EDTA
-
4C, Tris-HCl buffer, 50-100 mM Mg2+, Mn2+, Ba2+, or Ca2+, enzyme loses all activity in the presence of phosphate buffer or EDTA within 2 min
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
about 1000fold, to homogeneity from liver mitochondrial membranes by solubilization with Triton X-100, DEAE-ion-exchange chromatography and phenyl-resin chromatography
-
affinity chromatography on two triazine dyes
-
ammonium sulfate precipitation, DEAE-cellulose
-
ammonium sulfate precipitation, DEAE-Sephadex, Sephadex G-200
-
ammonium sulfate, acid precipitation, DEAE-Sepharose, glass beads
-
ammonium sulfate, DEAE-Sepharose, 60C, 10 min, hydroxylapatite, octyl-Sepharose, Sephadex G-200, DEAE-Sepharose
-
BDH3 partially purified from bdh2 mutant by three steps of column chromatography. Gene product of bdh3 expressed in Escherichia coli purified with 100% yield by two steps of column chromatography
charge-controlled hydrophobic chromatography and Sephadex G-100 gel filtration
-
DEAE-cellulose, Matrex gel blue A, Sephadex G-200, chromatofocusing
-
native by 3 chromatographic steps, recombinant from Escherichia coli in 2chromatographic steps
Acidovorax sp.
native enzyme by ammonium sulfate fractionation and affinity chromatography
-
phospholipase A2 treatment, controlled-pore glass beads
protamine sulfate precipitation, Sephacryl S-400, S200, DEAE-Sepharose
-
recombinant
-
recombinant C-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography
-
recombinant enzyme from Escherichia coli strain DH1 by anion exchange chromatography, hydrophobic interaction chromatography using a gradient of 1-20% ammonium sulfate, and ultrafiltration
-
wild-type and mutants purified by metal chelating affinity chromatography followed by ion-exchange chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
BDH3 expressed in Escherichia coli BLR (DE3)/pLysS harboring pETT13
DNA and amino acid sequence determination and analysis
-
DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain BL21(DE3)
Acidovorax sp.
efficiently expressed in Escherichia coli cells harboring pHBDH11
-
enzyme expression in Escherichia coli strain DH1
-
enzyme expression in Escherichia coli strain XL-1 Blue; expressed in Escherichia coli XL1-Blue cells
-
expression in Escherichia coli
-
expression in Escherichia coli; expression in Escherichia coli
expression of C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3)
-
functional co-expression with acetoacetyl-CoA thiolase and PHB synthase in Escherichia coli, recombinant production of poly(3-hydroxybutanoate) granules, overview
-
His-tagged 3-hydroxybutyrate dehydrogenase, expressed in Escherichia coli
-
mature form, expressed in Spodoptera frugiperda, SF9 cells
-
wild-type and mutants expressed in Escherichia coli XL1Blue harbouring pHBDH11. Wild-type HBDH and mutants containing an N-terminal His-tag expressed in Escherichia coli XL1Blue using pQE30 as a vector
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H144L/W187F
-
site-directed mutagenesis, the mutant shows activity with levulinnic acid, in contrast to the wild-type enzyme, and is engineered for production of 4-hydroxyvaleric acid, molecular docking simulation, overview
C242S
-
PCR derived cDNA clone
M92V
-
PCR derived cDNA clone
S24T
-
PCR derived cDNA clone
H144A
-
catalytic efficiency (kcat/Km) is 0.2% of the activity of wild-type HBDH
K152A
-
no activity
K152E
-
very low activity
K152Q
-
very low activity
K152R
-
retains a significant level of activity
L215A
-
both Km and kcat values are largely affected and the catalytic efficiency (kcat/Km) is less than 3% that of the wild-type enzyme
L215V
-
Km values increase 3.5- and 4.3fold and the kcat values are 73-118% those of the wild-type toward D-3-hydroxybutyrate and acetoacetate, respectively. Mutation does not significantly change Km and kcat toward NAD+ and NADH
Q196A
-
kcat/Km value is 0.6% that of the wild-type
Q196E
-
substantially reduced activity
Q196N
-
substantially reduced activity
Q94A
-
catalytic efficiency (kcat/Km) is 1.4% of the activity of wild-type HBDH
T190A
-
activity decreases to 0.1% that of the wild-type enzyme
T190C
-
decreased activity
T190S
-
retains 37% of the activity
W187A
-
very low activity
W187F
-
shows significant activity levels, 65% that of the wild-type enzyme
W187T
-
shows faint activity
W187Y
-
shows significant activity levels, 41% that of the wild-type enzyme
W257A
-
no activity
W257F
-
shows low activity levels, 2% that of the wild-type enzyme
W257Y
-
shows low activity levels, 1% that of the wild-type enzyme
Y155F
-
no activity
K149A
-
inactive mutant enzyme
K149R
-
kcat/KM for (R)-3-hydroxybutanoate is 184.2fold lower than wild-type value, kcat/Km for NAD+ is 7.2fold lower than wild-type enzyme
L149A
-
inactive
Q133A
-
decreased activity
Q193A
-
kcat/KM for (R)-3-hydroxybutanoate is 307fold lower than wild-type value, kcat/Km for NAD+ is 6.2fold lower than wild-type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
diagnostics
-
the enzyme is a useful marker in the assay of diabetes mellitus and/or ketoacidosis
synthesis
-
the engineered enzyme mutant H144L/W187F is used for production of 4-hydroxyvaleric acid, a monomer of bio-polyester and a precursor of bio-fuels, from levulinic acid
Show AA Sequence (1707 entries)
Please use the Sequence Search for a certain query.