Information on EC 3.5.1.19 - nicotinamidase

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

EC NUMBER
COMMENTARY hide
3.5.1.19
-
RECOMMENDED NAME
GeneOntology No.
nicotinamidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
nicotinamide + H2O = nicotinate + NH3
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
aldoxime degradation
-
-
Metabolic pathways
-
-
NAD metabolism
-
-
NAD salvage pathway I
-
-
Nicotinate and nicotinamide metabolism
-
-
pyridine nucleotide cycling (plants)
-
-
SYSTEMATIC NAME
IUBMB Comments
nicotinamide amidohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9033-32-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Borrelia burgdorferi
-
-
-
Manually annotated by BRENDA team
strain 544, ATCC 23448, gene pncA, a facultative intracellular pathogen
-
-
Manually annotated by BRENDA team
strain 544, ATCC 23448, gene pncA, a facultative intracellular pathogen
-
-
Manually annotated by BRENDA team
K12
-
-
Manually annotated by BRENDA team
Flavobacterium peregrinum
-
-
-
Manually annotated by BRENDA team
Leishmania infantum MHOM/MA/67/ITMAP-263
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
i.e. Micrococcus lysodeiktikus
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
deep-sea extremely halotolerant and alkaliphilic organism
UniProt
Manually annotated by BRENDA team
deep-sea extremely halotolerant and alkaliphilic organism
UniProt
Manually annotated by BRENDA team
strain GDI 211
-
-
Manually annotated by BRENDA team
strain GDI 211
-
-
Manually annotated by BRENDA team
strain CA401, strain FA18, strain FA64, strain FA86, strain FA131, strain 569B
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
-
nicotinamidases are a family of peptide hydrolases that generally contain a Zn2+ ion
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-methylnicotinamide + H2O
1-methylnicotinate + NH3
show the reaction diagram
-
-
-
-
?
4-methoxypyridine-3-carboxamide + H2O
?
show the reaction diagram
5-methoxypyridine-3-carboxamide + H2O
?
show the reaction diagram
5-methyl nicotinamide + H2O
5-methyl nicotinate + NH3
show the reaction diagram
5-methylnicotinamide + H2O
5-methylnicotinate + NH3
show the reaction diagram
5-methylpyridine-3-carboxamide + H2O
?
show the reaction diagram
6-aminopyridine-3-carboxamide + H2O
?
show the reaction diagram
6-chloropyridine-3-carboxamide + H2O
?
show the reaction diagram
6-methylpyridine-3-carboxamide + H2O
?
show the reaction diagram
benzamide + H2O
benzoate + NH3
show the reaction diagram
-
-
-
-
?
ethyl nicotinate + H2O
nicotinate + ethanol
show the reaction diagram
-
-
-
?
ethyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
methyl nicotinate + H2O
nicotinate + methanol
show the reaction diagram
-
-
-
?
methyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
nicotinamide + H2O
NH3 + nicotinic acid
show the reaction diagram
nicotinamide + H2O
nicotinate + NH3
show the reaction diagram
nicotinamide + H2O
nicotinic acid + NH3
show the reaction diagram
p-nitrophenol + H2O
phenol + NH3
show the reaction diagram
-
-
-
-
?
p-nitrophenyl nicotinate + H2O
?
show the reaction diagram
-
-
-
-
?
phenyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
pyrazinamide + H2O
NH3 + pyrazinoic acid
show the reaction diagram
pyrazinamide + H2O
pyrazine-2-carboxylic acid + NH3
show the reaction diagram
pyrazinamide + H2O
pyrazinic acid + NH3
show the reaction diagram
pyrazinamide + H2O
pyrazinoic acid + NH3
show the reaction diagram
pyridine-3-carbothioamide + H2O
?
show the reaction diagram
-
44% activity compared to nicotinamide
-
-
?
thionicotinamide + H2O
pyridine-3-carbothioic O-acid
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
4-methoxypyridine-3-carboxamide + H2O
?
show the reaction diagram
5-methoxypyridine-3-carboxamide + H2O
?
show the reaction diagram
5-methylpyridine-3-carboxamide + H2O
?
show the reaction diagram
6-aminopyridine-3-carboxamide + H2O
?
show the reaction diagram
6-chloropyridine-3-carboxamide + H2O
?
show the reaction diagram
6-methylpyridine-3-carboxamide + H2O
?
show the reaction diagram
ethyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
methyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
nicotinamide + H2O
NH3 + nicotinic acid
show the reaction diagram
nicotinamide + H2O
nicotinate + NH3
show the reaction diagram
nicotinamide + H2O
nicotinic acid + NH3
show the reaction diagram
phenyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
pyrazinamide + H2O
NH3 + pyrazinoic acid
show the reaction diagram
pyrazinamide + H2O
pyrazine-2-carboxylic acid + NH3
show the reaction diagram
pyrazinamide + H2O
pyrazinic acid + NH3
show the reaction diagram
pyrazinamide + H2O
pyrazinoic acid + NH3
show the reaction diagram
-
a nicotinamide prodrug. In Mycobacterium tuberculosis the enzyme converts the nicotinamide analogue prodrug pyrazinamide into the bacteriostatic pyrazinoic acid, hence the alternative name, pyrazinamidase, PncA
a bacteriostatic drug
-
?
pyridine-3-carbothioamide + H2O
?
show the reaction diagram
-
44% activity compared to nicotinamide
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
activation
Ni2+
-
enhances activity
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-dipyridyl
-
Mg2+ reverses inhibition
2-chloropyridine-3-carbaldehyde
2-mercaptoethanol
-
-
3-(3-pyridyl)-1-propanol
-
moderate
3-acetylpyridine
3-aminomethylpyridine
-
potent
3-chloropyridine
-
moderate
3-Cyanopyridine
3-hydroxypyridine
-
-
3-indolepropionic acid
-
inhibits amidase and esterase activity
3-pyridine carboxaldehyde
-
competitive inhibitor
3-Pyridineacetonitrile
-
moderate
3-pyridinealdoxime
-
potent
3-Pyridinecarboxaldehyde
-
potent
3-Pyridylcarbinol
-
potent
4-aminonicotinamide
-
potent
4-methoxylnicotinaldehyde
5-bromonicotinaldehyde
competitive inhibition
-
5-bromopyridine-3-carbaldehyde
5-methoxynicotinaldehyde
Q97PM2
competitive inhibitor
5-methoxypyridine-3-carbaldehyde
5-methylnicotinaldehyde
5-Methylnicotinamide
5-methylpyridine-3-carbaldehyde
6-fluoropyridine-3-carbaldehyde
8-hydroxyquinoline
-
-
benzaldehyde
-
competitive inhibition
Benzoic acid
-
inhibits amidase and esterase activity
Carbobenzoxyamido-2-phenyl-ethyl-chloromethyl-ketone
-
-
diisopropylfluorophosphate
-
-
dithiothreitol
-
-
Fe3+
1 mM, 94% inhibition
guanidinium hydrochloride
-
3.4 M, complete inhibition
HgCl2
Flavobacterium peregrinum
-
-
Hydrocinnamic acid
-
inhibits amidase and esterase activity
iodoacetamide
-
reversible, competitive inhibitor
Isoniazid
-
moderate
L-thyroxine
-
-
N,N'-diethylnicotinamide
Flavobacterium peregrinum
-
competitive
N-ethylmaleimide
-
-
N2-ethylnicotinamide
Flavobacterium peregrinum
-
competitive
N2-methylnicotinamide
Flavobacterium peregrinum
-
competitive
NAD+
-
competitive with respect to nicotinamide
nicotinal hydroxamic acid
-
moderate
nicotinaldehyde
nicotinamide
-
inhibits esterolysis of p-nitrophenyl acetate
nicotinamide analogs
Flavobacterium peregrinum
-
analogs with a trivalent nitrogen, competitive inhibition
-
nicotinic acid
nicotinic acid hydrazide
nipecotamide
-
moderate
o-bromobenzamide
-
moderate
p-nitrophenol
-
inhibits amidase and esterase activity
Pyrazinamide
pyrazinecarbonitrile
-
irreversible inactivator
pyrazinoic acid
-
competitive inhibition
pyridine-3-carbonitrile
-
reversible, competitive inhibitor
Sodium diethyl dithiocarbamate
-
-
sulfhydryl reagents
-
-
Thionicotinamide
-
moderate
Triton X-100
-
0.1%, 24% inhibition
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
dithiothreitol
Serum albumin
-
0.02%, 9% activation
-
additional information
-
stress suppresses the inhibitory effect of nicotinamide through the induction of PNC1 expression
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.36
5'-methylnicotinamide
-
-
0.55 - 0.68
5-methyl nicotinamide
0.061
5-Methylnicotinamide
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25C
0.025
Benzamide
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25C
0.34 - 1.43
ethyl nicotinate
0.44 - 1.03
methyl nicotinate
0.0002 - 50
nicotinamide
0.016 - 1.2
Pyrazinamide
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.16 - 15.9
5-methyl nicotinamide
1.75
5-Methylnicotinamide
Saccharomyces cerevisiae
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25C
0.0088
Benzamide
Saccharomyces cerevisiae
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25C
0.05 - 0.19
ethyl nicotinate
0.17 - 1.02
methyl nicotinate
0.0005 - 76.9
nicotinamide
0.1 - 135.7
Pyrazinamide
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0086
1-methylnicotinamide
Saccharomyces cerevisiae
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25C
12283
3.4 - 29.04
5-methyl nicotinamide
0.061 - 29
5-Methylnicotinamide
0.025 - 0.35
Benzamide
0.05 - 0.41
ethyl nicotinate
0.31 - 1.17
methyl nicotinate
0.0096 - 427
nicotinamide
0.157 - 331
Pyrazinamide
0.05
Thionicotinamide
Saccharomyces cerevisiae
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25C
8199
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.37 - 5
2-chloropyridine-3-carbaldehyde
0.316
3-acetylpyridine
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25C
0.085 - 2
3-Cyanopyridine
0.00029
3-pyridine carboxaldehyde
-
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25C
0.00044 - 0.153
4-methoxylnicotinaldehyde
0.0044
5-bromonicotinaldehyde
recombinant wild-type enzyme, pH 7.3, 37C
-
0.00059 - 0.004
5-bromopyridine-3-carbaldehyde
0.000039 - 0.0038
5-methoxypyridine-3-carbaldehyde
0.000023
5-methylnicotinaldehyde
0.00019 - 0.00065
5-methylpyridine-3-carbaldehyde
0.004 - 0.018
6-fluoropyridine-3-carbaldehyde
0.0206
benzaldehyde
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25C
0.000011 - 0.0035
nicotinaldehyde
0.12 - 2
nicotinic acid
6.7
pyrazinoic acid
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25C
0.1
pyridine-3-carbonitrile
-
Ki about 0.1 mM, wild type enzyme, in 100 mM HEPES at pH 8.0 and 25C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.06
purified recombinant mutant Q96K, pH 7.3, 37C, substrate ethyl nicotinate
0.13
purified recombinant wild-type enzyme, pH 7.3, 37C, substrate ethyl nicotinate
0.22
purified recombinant mutant Q96A, pH 7.3, 37C, substrate ethyl nicotinate
0.31
purified recombinant mutant Q96K, pH 7.3, 37C, substrate methyl nicotinate
0.34
purified recombinant mutant C133A, pH 7.3, 37C, substrate ethyl nicotinate
0.36
purified recombinant mutant K104A, pH 7.3, 37C, substrate nicotinamide
0.37
purified recombinant mutant T12Q, pH 7.3, 37C, substrate nicotinamide
0.45
purified recombinant wild-type enzyme, pH 7.3, 37C, substrate methyl nicotinate
0.46
purified recombinant mutant Q96K, pH 7.3, 37C, substrate pyrazinamide
0.7
purified recombinant mutant E65H, pH 7.3, 37C, substrate pyrazinamide
1.16
purified recombinant mutant Q96A, pH 7.3, 37C, substrate methyl nicotinate
1.4
purified recombinant mutant C133A, pH 7.3, 37C, substrate methyl nicotinate
1.8
purified recombinant mutant Q96A, pH 7.3, 37C, substrate pyrazinamide
2.17
Flavobacterium peregrinum
-
-
3.62
purified recombinant wild-type enzyme, pH 7.3, 37C, substrate pyrazinamide
3.96
purified recombinant mutant Q96K, pH 7.3, 37C, substrate 5-methyl nicotinamide
6.39
purified recombinant mutant C133A, pH 7.3, 37C, substrate pyrazinamide
7.43
purified recombinant mutant F68W/C133A, pH 7.3, 37C, substrate pyrazinamide
7.5
purified recombinant mutant F68W, pH 7.3, 37C, substrate pyrazinamide
8.7
purified recombinant mutant Q96K, pH 7.3, 37C, substrate nicotinamide
9.01
purified recombinant wild-type enzyme, pH 7.3, 37C, substrate 5-methyl nicotinamide
11.74
purified recombinant mutant C133A, pH 7.3, 37C, substrate 5-methyl nicotinamide
13.5
purified recombinant mutant E65H, pH 7.3, 37C, substrate nicotinamide
17.5
purified recombinant mutant C133A, pH 7.3, 37C, substrate nicotinamide
19.97
purified recombinant mutant F68W, pH 7.3, 37C, substrate nicotinamide
22.9
purified recombinant mutant F68W/C133A, pH 7.3, 37C, substrate nicotinamide
23.3 - 25
purified recombinant wild-type enzyme, pH 7.3, 37C, substrate nicotinamide
24.8
purified recombinant mutant Q96A, pH 7.3, 37C, substrate nicotinamide
28
purified recombinant mutant Q96A, pH 7.3, 37C, substrate 5-methyl nicotinamide
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
Flavobacterium peregrinum
-
-
6.5 - 8.5
-
amidase activity
6.6
Flavobacterium peregrinum
-
-
6.8 - 7.5
-
-
8 - 10
-
esterase
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
activity range, marked decrease in activity below pH 5.0 and above pH 8.0
5 - 10.5
-
pH 5.0: about 70% of maximal activity, pH 10.5: about 75% of maximal activity
5.5 - 9.5
-
pH 5.5: about 60% of maximal activity, pH 9.5: about 45% of maximal activity
6.5 - 10
-
hydrolysis of nicotinamide of a wide range of pH values
7 - 11
-
pH 7.0: about 35% of maximal activity, pH 11.0: about 30% of maximal activity
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 55
activity range, below 25C or above 55C the enzyme loses most of its activity
55 - 80
-
about 35% of maximal activity at 55C and at 80C
60 - 100
-
60C: about 45% of maximal activity, 100C: 85% of maximal activity
65 - 98
-
65C: about 50% of maximal activity, 98C: about 80% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
C1300 neuroblastoma M1 clonal cell line
Manually annotated by BRENDA team
-
embryonic axis
Manually annotated by BRENDA team
-
human erythrocytes infected with Plasmodium falciparum
Manually annotated by BRENDA team
-
activity during germination, overview
Manually annotated by BRENDA team
-
highest level of expression
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
surface location
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Acinetobacter baumannii (strain AYE)
Acinetobacter baumannii (strain AYE)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20890
-
determined by analytical ultracentrifugation and mass spectrometry
39700
dimeric enzyme, gel filtration
42900
gel filtration
43000
-
gel filtration
47000
-
gel filtratrion and analytical ultracentrifugation
48000
Flavobacterium peregrinum
-
gel filtration
211000
-
gel filtration
230000
-
gel electrophoresis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
homotetramer
Q97PM2
4 * 21400, calculated from amino acid sequence
monomer
tetramer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
contains 1.7% w/w mannose and 1.5% w/w N-acetylglucosamine
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure (PDB ID 2WT9) analysis, and modeling of substrate binding and unbinding, overview
sitting-drop-based and sparse-matrix screening , using 0.7 M Na-citrate and 0.1 M HEPES (pH 7.5)
-
in complex with nicotinaldehyde, hanging drop vapor diffusion method, using 1.6 M NaOAc, 10% (w/v) ethylene glycol, and 0.1 M HEPES (pH 7.4)
-
resolution 2.9 Angstrom
-
a trapped nicotinoyl-thioester complexed with wild type enzyme and C136S mutant in complex with nicotinamide, hanging drop vapor diffusion method, using 18-22% (w/v) polyethylene glycol 3350, 0.2-0.3 M NaCl and 0.2 M sodium malonate, pH 6.3, at 18C
Q97PM2
crystal structure (PDB ID 3O94) analysis, and modeling of substrate binding and unbinding, overview
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
-
37C, 1 h, stable
246570
6 - 8
-
-
679806
6 - 7.3
purified recombinant wild-type enzyme, very stable at pH 6.0 and pH 7.3, where it maintains 40% and 30% residual activity, respectively, after 20 hours of incubation
735084
7 - 9.5
-
37C, 10 min, stable
246573
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 20
purified recombinant wild-type enzyme, 20 h, stable
40
-
pH 6.8, in presence of Co2+, 10 min, stable up to
55
-
half-life: 2.5 min
65
-
half-life: 1 min
70
-
10 min, complete loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme is stable in buffer solution containing Co2+ and Ni2+
-
freezing and defreezing inactivates
-
inactivated by dialysis in absence of Mn2+. After overnight dialysis in presence of Mn2+ almost all activity is retained. A dialysis against 0.01 M maleate buffer, pH 6.5, containing 0.005 mM HgCl2 results in complete retention of activity
Flavobacterium peregrinum
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
photooxidation of two His residues by methylene blue leads to loss of deaminase activity
-
246572
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 50% glycerol, stable for months
-
-20C, 50% loss of activity after 4 weeks
Flavobacterium peregrinum
-
0C, phosphate buffer, pH 7.0, weak loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
immobilized metal affinity chromatography
-
Ni-NTA agarose column chromatography
-
Ni-NTA column chromatography
-
Ni-NTA column chromatography and Sephacryl S-300 gel filtration
-
Ni-NTA resin column chromatography
Q97PM2
nickel chelate chromatography and molecular sieve
-
nickel-based affinity chromatography
-
recombinant enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography, ultrafiltration, and gel filtration
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes methylnicotinamide from Escherichia coli strain Rosetta 2 (DE3) by ultrafiltration and nickel affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a single enzyme accounts for both pyrazinamidase activity and nicotinamidase activity
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli Rosetta 2(DE3) pLysS cells
Q97PM2
expression in Escherichia coli
gene PNC1, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
gene pncA, DNA and amino acid sequence determination and analysis, recombinant enzyme expression in Escherichia coli strain BL21(DE3)pLysS
isolation of a pnc-1a promoter consisting of exon 1a and the intervening region between exon 1a and the adjacent gene, expression as GFP-fusion protein. Expression of His-tagged PNC proteins in Escherichia coli strain BL21(DE3)
-
phylogenetic analysis, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta 2 (DE3)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C138A
-
significant decrease in enzyme activity
D49A
-
significant decrease in enzyme activity
D8A
-
significant decrease in enzyme activity
H51A
-
significant decrease in enzyme activity
H57A
-
significant decrease in enzyme activity
H57D
-
the mutant has reduced Mn2+ content and shows a 6fold and 38fold decrease in kcat value for nicotinamide and pyrazinamide, respectively
H71A
-
significant decrease in enzyme activity
K96A
-
significant decrease in enzyme activity
S104A
-
partial loss of enzyme activity
S95A
-
partial loss of enzyme activity
H57D
-
the mutant has reduced Mn2+ content and shows a 6fold and 38fold decrease in kcat value for nicotinamide and pyrazinamide, respectively
-
C133A
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
E65H
site-directed mutagenesis, the mutation highly reduces the activity with pyrazinamide compared to the wild-type enzyme
F68W
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
F68W/C133A
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
K104A
site-directed mutagenesis, the mutant shows 98.55% reduced activity compared to the wild-type enzyme
Q96A
site-directed mutagenesis, the mutant shows similar activity with nicotinamide and increased activity with 5-methyl nicotinamide compared to the wild-type enzyme
Q96K
site-directed mutagenesis, the mutant shows 62.7% reduced activity compared to the wild-type enzyme
T12Q
site-directed mutagenesis, the mutant shows 98.42% reduced activity compared to the wild-type enzyme
C133A
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site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
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K104A
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site-directed mutagenesis, the mutant shows 98.55% reduced activity compared to the wild-type enzyme
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Q96A
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site-directed mutagenesis, the mutant shows similar activity with nicotinamide and increased activity with 5-methyl nicotinamide compared to the wild-type enzyme
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T12Q
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site-directed mutagenesis, the mutant shows 98.42% reduced activity compared to the wild-type enzyme
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D51A
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the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme
D51N
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the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme
D8A
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the mutant shows 770fold decreased kcat value compared to the wild type enzyme
D8E
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the mutant shows 100fold decreased kcat value compared to the wild type enzyme
D8N
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the mutant shows 1000fold decreased kcat value compared to the wild type enzyme
H53A
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the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme
H94A
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the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme
K122A
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the mutant shows 16fold decreased kcat value compared to the wild type enzyme
K122R
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the mutant shows 770fold decreased kcat value compared to the wild type enzyme
C136A
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the mutant is unable to catalyze nicotinamide hydrolysis
K103A
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the mutant has only 0.15% of the catalytic activity of the native enzyme
R97A
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the mutant is a robust nicotinamidase and is nearly as good as the native enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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determination of nicotinamide
drug development
Show AA Sequence (1186 entries)
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