BRENDA - Enzyme Database
show all sequences of 2.7.7.47

Characterization of the bifunctional aminoglycoside-modifying enzyme ANT(3)-Ii/AAC(6)-IId from Serratia marcescens

Kim, C.; Hesek, D.; Zajicek, J.; Vakulenko, S.B.; Mobashery, S.; Biochemistry 45, 8368-8377 (2006)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
cloned in Escherichia coli BL21
Serratia marcescens
Inhibitors
Inhibitors
Commentary
Organism
Structure
9-O-(adenosine-5'-phophoryl)spectinomycin
product inhibition of ANT(3'')-Ii domain
Serratia marcescens
AMP-CPP
dead-end inhibition of ANT(3'')-Ii domain
Serratia marcescens
kanamycin A
dead-end inhibition of ANT(3'')-Ii domain
Serratia marcescens
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0013
-
streptomycin
kinetic parameter of the ANT(3'')-Ii domain of the enzyme
Serratia marcescens
0.0014
-
spectinomycin
kinetic parameter of the ANT(3'')-Ii domain of the enzyme
Serratia marcescens
0.014
-
ATP
kinetic parameter of the ANT(3'')-Ii domain of the enzyme
Serratia marcescens
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
assay with 15 mM MgCl2
Serratia marcescens
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Serratia marcescens
-
-
-
Purification (Commentary)
Commentary
Organism
DEAE anion-exchange column and gentamycin affinity column
Serratia marcescens
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + spectinomycin
-
672194
Serratia marcescens
diphosphate + 9-adenylylspectinomycin
-
-
-
-
ATP + streptomycin
bifunctional enzyme, adenylation of aminoglycoside antibiotics takes place at the ANT(3'')-Ii domain, aceylation of aminoglycoside antibiotics takes place at the AAC(6')-domain
672194
Serratia marcescens
diphosphate + 3''-adenylylstreptomycin
-
-
-
?
additional information
adenyltransferase domain is highly specific for spectinomycin and streptomycin and catalyzes the reaction by a Theorell-Chance kinetic mechanism, where ATP binds to the enzyme prior to the aminoglycoside and the modified antibiotic is the last product to be released
672194
Serratia marcescens
?
-
-
-
-
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
additional information
-
assay is performed at room temperature
Serratia marcescens
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.5
-
ATP
turnover number of ANT(3'')-Ii domain
Serratia marcescens
0.5
-
spectinomycin
turnover number of ANT(3'')-Ii domain
Serratia marcescens
0.6
-
streptomycin
turnover number of ANT(3'')-Ii domain
Serratia marcescens
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Serratia marcescens
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.004
-
kanamycin A
dead-end inhibition of ANT(3'')-Ii domain, substrate 0.02 mM spectinomycin, competitive inhibition
Serratia marcescens
0.01
-
kanamycin A
Kii = 0.01 mM, dead-end inhibition of ANT(3'')-Ii domain, substrate 0.1 mM ATP, uncompetitive inhibition
Serratia marcescens
0.017
-
AMP-CPP
dead-end inhibition of ANT(3'')-Ii domain, substrate 0.1 mM ATP, competitive inhibition
Serratia marcescens
0.021
-
AMP-CPP
dead-end inhibition of ANT(3'')-Ii domain, substrate 0.02 mM spectinomycin, Kii = 0.031 mM, noncompetitive/mixed inhibition
Serratia marcescens
0.076
-
9-O-(adenosine-5'-phophoryl)spectinomycin
Kii = 0.089, product inhibition of ANT(3'')-Ii domain, substrate 0.02 mM spectinomycin, noncompetitive/mixed inhibition
Serratia marcescens
0.093
-
9-O-(adenosine-5'-phophoryl)spectinomycin
product inhibition of ANT(3'')-Ii domain, substrate 0.1 mM ATP, competitive inhibition
Serratia marcescens
Cloned(Commentary) (protein specific)
Commentary
Organism
cloned in Escherichia coli BL21
Serratia marcescens
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
9-O-(adenosine-5'-phophoryl)spectinomycin
product inhibition of ANT(3'')-Ii domain
Serratia marcescens
AMP-CPP
dead-end inhibition of ANT(3'')-Ii domain
Serratia marcescens
kanamycin A
dead-end inhibition of ANT(3'')-Ii domain
Serratia marcescens
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.004
-
kanamycin A
dead-end inhibition of ANT(3'')-Ii domain, substrate 0.02 mM spectinomycin, competitive inhibition
Serratia marcescens
0.01
-
kanamycin A
Kii = 0.01 mM, dead-end inhibition of ANT(3'')-Ii domain, substrate 0.1 mM ATP, uncompetitive inhibition
Serratia marcescens
0.017
-
AMP-CPP
dead-end inhibition of ANT(3'')-Ii domain, substrate 0.1 mM ATP, competitive inhibition
Serratia marcescens
0.021
-
AMP-CPP
dead-end inhibition of ANT(3'')-Ii domain, substrate 0.02 mM spectinomycin, Kii = 0.031 mM, noncompetitive/mixed inhibition
Serratia marcescens
0.076
-
9-O-(adenosine-5'-phophoryl)spectinomycin
Kii = 0.089, product inhibition of ANT(3'')-Ii domain, substrate 0.02 mM spectinomycin, noncompetitive/mixed inhibition
Serratia marcescens
0.093
-
9-O-(adenosine-5'-phophoryl)spectinomycin
product inhibition of ANT(3'')-Ii domain, substrate 0.1 mM ATP, competitive inhibition
Serratia marcescens
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0013
-
streptomycin
kinetic parameter of the ANT(3'')-Ii domain of the enzyme
Serratia marcescens
0.0014
-
spectinomycin
kinetic parameter of the ANT(3'')-Ii domain of the enzyme
Serratia marcescens
0.014
-
ATP
kinetic parameter of the ANT(3'')-Ii domain of the enzyme
Serratia marcescens
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
assay with 15 mM MgCl2
Serratia marcescens
Purification (Commentary) (protein specific)
Commentary
Organism
DEAE anion-exchange column and gentamycin affinity column
Serratia marcescens
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + spectinomycin
-
672194
Serratia marcescens
diphosphate + 9-adenylylspectinomycin
-
-
-
-
ATP + streptomycin
bifunctional enzyme, adenylation of aminoglycoside antibiotics takes place at the ANT(3'')-Ii domain, aceylation of aminoglycoside antibiotics takes place at the AAC(6')-domain
672194
Serratia marcescens
diphosphate + 3''-adenylylstreptomycin
-
-
-
?
additional information
adenyltransferase domain is highly specific for spectinomycin and streptomycin and catalyzes the reaction by a Theorell-Chance kinetic mechanism, where ATP binds to the enzyme prior to the aminoglycoside and the modified antibiotic is the last product to be released
672194
Serratia marcescens
?
-
-
-
-
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
additional information
-
assay is performed at room temperature
Serratia marcescens
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.5
-
ATP
turnover number of ANT(3'')-Ii domain
Serratia marcescens
0.5
-
spectinomycin
turnover number of ANT(3'')-Ii domain
Serratia marcescens
0.6
-
streptomycin
turnover number of ANT(3'')-Ii domain
Serratia marcescens
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Serratia marcescens
Other publictions for EC 2.7.7.47
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)
737356
Chen
Structure of AadA from Salmone ...
Salmonella enterica subsp. enterica serovar Typhimurium, Salmonella enterica subsp. enterica serovar Typhimurium LT2
Acta Crystallogr. Sect. D
71
2267-2277
2015
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1
8
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2
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4
1
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1
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1
8
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4
1
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1
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-
739973
Papadovasilaki
Biophysical and enzymatic prop ...
Pseudomonas aeruginosa, Pseudomonas aeruginosa Ps100
Biochem. Biophys. Rep.
4
152-157
2015
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1
1
1
-
2
6
-
1
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2
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1
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1
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6
2
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-
2
6
-
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1
-
1
1
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2
-
6
-
1
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1
-
-
1
-
6
2
-
-
2
6
-
-
-
-
-
-
-
-
6
6
723011
Shahi
Interaction of dihydrofolate r ...
Klebsiella pneumoniae
J. Mol. Model.
19
973-983
2013
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1
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4
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706857
Singh
Plastid transformation in eggp ...
Escherichia coli
Transgenic Res.
19
113-119
2010
-
1
1
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4
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1
1
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701575
Ahmed
Genetic basis of multidrug res ...
Salmonella enterica subsp. enterica serovar Enteritidis, Salmonella enterica subsp. enterica serovar Typhimurium
Acta Trop.
111
144-149
2009
-
2
-
-
-
-
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6
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2
2
-
-
-
703283
Ajiboye
Global spread of mobile antimi ...
Escherichia coli, Salmonella enterica subsp. enterica serovar Typhimurium
Clin. Infect. Dis.
49
365-371
2009
-
2
-
-
-
-
-
-
-
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16
-
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10
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-
-
-
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703804
Mathew
Evidence of class 1 integron t ...
Escherichia coli, Salmonella
Foodborne Pathog. Dis.
6
959-964
2009
-
-
2
-
-
-
-
-
-
-
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-
7
-
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2
2
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704129
Randhawa
Multiplex PCR-based simultaneo ...
Gossypium hirsutum
J. Agric. Food Chem.
57
5167-5172
2009
-
1
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1
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1
1
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706886
Ahmed
Genetic analysis of antimicrob ...
Escherichia coli
Vet. Microbiol.
136
397-402
2009
-
-
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-
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3
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1
1
-
-
-
672194
Kim
Characterization of the bifunc ...
Serratia marcescens
Biochemistry
45
8368-8377
2006
-
-
1
-
-
-
3
3
-
1
-
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1
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1
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3
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1
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3
1
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6
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1
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3
6
3
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1
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1
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3
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1
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3
1
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672299
Jana
Effects of guanidine hydrochlo ...
Escherichia coli
Biochemistry (Moscow)
71
1230-1237
2006
-
-
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1
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1
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1
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1
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1
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1
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675323
Chen
Insertion sequence ISEcp1-like ...
Enterococcus casseliflavus, Enterococcus casseliflavus HZ95
J. Med. Microbiol.
55
1521-1525
2006
-
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1
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1
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8
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661382
Jana
Kinetic mechanism of streptomy ...
Escherichia coli
Biotechnol. Lett.
27
519-524
2005
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2
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663311
Jana
Purification of streptomycin a ...
Escherichia coli
Protein Expr. Purif.
40
86-90
2005
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1
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2
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1
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1
1
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643347
Tauch
The 27.8-kb R-plasmid pTET3 fr ...
Corynebacterium glutamicum, Corynebacterium glutamicum LP-6
Plasmid
48
117-129
2002
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1
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1
2
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8
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6
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1
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1
2
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6
-
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643346
Clark
Detection of a streptomycin/sp ...
Enterococcus faecalis
Antimicrob. Agents Chemother.
43
157-160
1999
-
1
1
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1
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1
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4
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1
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1
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4
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643345
Kazama
A new gene, aadA2b, encoding a ...
Escherichia coli, Pseudomonas aeruginosa
Microbios
86
77-83
1996
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1
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1
2
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2
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4
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