BRENDA - Enzyme Database

Purification and properties of two gentamicin-modifying enzymes, coded by a single plasmid pPK237 originating from Pseudomonas aeruginosa

Angelatou, F.; Litsas, S.B.; Kontomichalou, P.; J. Antibiot. 35, 235-244 (1982)

Data extracted from this reference:

Application
EC Number
Application
Commentary
Organism
2.7.7.46
medicine
gentamicin resistance, mediated by multiple resistance plasmids in different species of Enterobacteriaceae, can be a problem in hospitals
Pseudomonas aeruginosa
Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
2.3.1.81
Pseudomonas aeruginosa plasmid pPK237 transferred to and expressed in Escherichia coli K12
Pseudomonas aeruginosa
General Stability
EC Number
General Stability
Organism
2.3.1.81
dialysis against EDTA with subsequent Mg2+-addition to dialyzed enzyme has no effect on activity
Pseudomonas aeruginosa
2.3.1.81
gel filtration leads to complete inactivation
Pseudomonas aeruginosa
2.7.7.46
highly unstable after purification
Pseudomonas aeruginosa
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.1.81
EDTA
above 0.5 mM
Pseudomonas aeruginosa
2.3.1.81
additional information
no inhibition by NH4+, K+, Na+, DTT, N-methylmaleimide
Pseudomonas aeruginosa
2.3.1.81
Zn2+
-
Pseudomonas aeruginosa
2.7.7.46
EDTA
above 0.5 mM
Pseudomonas aeruginosa
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.1.81
0.066
-
gentamicin C1
-
Pseudomonas aeruginosa
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.3.1.81
Ca2+
1.0 mM, about 100% activity enhancement
Pseudomonas aeruginosa
2.3.1.81
Mg2+
1.5 mM, about 100% activity enhancement
Pseudomonas aeruginosa
2.3.1.81
NH4+
slight activation
Pseudomonas aeruginosa
2.7.7.46
Ca2+
shows enzymatic activity only in presence of cations, optimal concentration 1.0 mM
Pseudomonas aeruginosa
2.7.7.46
Mg2+
shows enzymatic activity only in presence of cations, optimal concentration 2.0 mM
Pseudomonas aeruginosa
2.7.7.46
additional information
K+ and Na+ have almost no effect, NH4+ has a low enhancing effect
Pseudomonas aeruginosa
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.1.81
additional information
-
amino acid analysis
Pseudomonas aeruginosa
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2.7.7.46
ATP + gentamicin A
Pseudomonas aeruginosa
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
diphosphate + 2''-adenylylgentamicin A
-
Pseudomonas aeruginosa
r
Organism
EC Number
Organism
UniProt
Commentary
Textmining
2.3.1.81
Pseudomonas aeruginosa
-
plasmid pPK237 transferred to and expressed in Escherichia coli K12
-
2.7.7.46
Pseudomonas aeruginosa
-
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
2.3.1.81
plasmid pPK237 transferred to and expressed in E. coli K12, affinity chromatography
Pseudomonas aeruginosa
2.7.7.46
plasmid pPK237, purified from an E. coli K12 host RC85F-
Pseudomonas aeruginosa
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
2.3.1.81
34
-
-
Pseudomonas aeruginosa
Storage Stability
EC Number
Storage Stability
Organism
2.3.1.81
-20C, partially purified, several months
Pseudomonas aeruginosa
2.3.1.81
-20C, purified, several days
Pseudomonas aeruginosa
2.7.7.46
4C, activity remains only for a few hours
Pseudomonas aeruginosa
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2.3.1.81
acetyl-CoA + dibekacin
acetylation at 8% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3''-acetyldibekacin
-
-
-
?
2.3.1.81
acetyl-CoA + gentamicin A
acetylation at 6% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin A
-
-
-
?
2.3.1.81
acetyl-CoA + gentamicin C
acetylation at 86% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin C
-
-
-
?
2.3.1.81
acetyl-CoA + gentamicin C1
best substrate
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin C1
-
-
-
?
2.3.1.81
acetyl-CoA + gentamicin C1a
acetylation at 60% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin C1a
-
637300
Pseudomonas aeruginosa
?
2.3.1.81
acetyl-CoA + gentamicin C2
acetylation at 84% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin C2
-
-
-
?
2.3.1.81
acetyl-CoA + kanamycin A
acetylation at 7% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylkanamycin A
-
-
-
?
2.3.1.81
acetyl-CoA + sisomicin
acetylation at 51% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylsisomicin
-
-
-
?
2.3.1.81
acetyl-CoA + tobramycin
acetylation at 6% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetyltobramycin
-
-
-
?
2.3.1.81
additional information
no substrates are amikacin
637300
Pseudomonas aeruginosa
?
-
-
-
-
2.3.1.81
additional information
lividomycin A, butirosin A and B
637300
Pseudomonas aeruginosa
?
-
-
-
-
2.7.7.46
ATP + dibekacin
-
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylyldibekacin
-
-
-
r
2.7.7.46
ATP + gentamicin A
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylgentamicin A
-
637300
Pseudomonas aeruginosa
r
2.7.7.46
ATP + gentamicin C1
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylgentamicin C1
-
-
-
-
2.7.7.46
ATP + gentamicin C1a
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylgentamicin C1a
-
-
-
-
2.7.7.46
ATP + gentamicin C2
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylgentamicin C2
-
-
-
-
2.7.7.46
ATP + kanamycin A
poor substrate
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylkanamycin A
-
-
-
r
2.7.7.46
ATP + sisomicin
-
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylsisomicin
-
-
-
r
2.7.7.46
ATP + tobramycin
poor substrate
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylyltobramycin
-
-
-
r
2.7.7.46
additional information
amikacin, neomycin, lividomycin and butirosin A and B are no substrates
637300
Pseudomonas aeruginosa
?
-
-
-
-
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
2.3.1.81
37
-
assay at
Pseudomonas aeruginosa
Temperature Stability [C]
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
2.3.1.81
40
-
above, inactivation within 2 min
Pseudomonas aeruginosa
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.1.81
7
7.6
-
Pseudomonas aeruginosa
2.7.7.46
7.9
-
-
Pseudomonas aeruginosa
Application (protein specific)
EC Number
Application
Commentary
Organism
2.7.7.46
medicine
gentamicin resistance, mediated by multiple resistance plasmids in different species of Enterobacteriaceae, can be a problem in hospitals
Pseudomonas aeruginosa
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.1.81
Pseudomonas aeruginosa plasmid pPK237 transferred to and expressed in Escherichia coli K12
Pseudomonas aeruginosa
General Stability (protein specific)
EC Number
General Stability
Organism
2.3.1.81
dialysis against EDTA with subsequent Mg2+-addition to dialyzed enzyme has no effect on activity
Pseudomonas aeruginosa
2.3.1.81
gel filtration leads to complete inactivation
Pseudomonas aeruginosa
2.7.7.46
highly unstable after purification
Pseudomonas aeruginosa
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.1.81
EDTA
above 0.5 mM
Pseudomonas aeruginosa
2.3.1.81
additional information
no inhibition by NH4+, K+, Na+, DTT, N-methylmaleimide
Pseudomonas aeruginosa
2.3.1.81
Zn2+
-
Pseudomonas aeruginosa
2.7.7.46
EDTA
above 0.5 mM
Pseudomonas aeruginosa
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.1.81
0.066
-
gentamicin C1
-
Pseudomonas aeruginosa
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.3.1.81
Ca2+
1.0 mM, about 100% activity enhancement
Pseudomonas aeruginosa
2.3.1.81
Mg2+
1.5 mM, about 100% activity enhancement
Pseudomonas aeruginosa
2.3.1.81
NH4+
slight activation
Pseudomonas aeruginosa
2.7.7.46
Ca2+
shows enzymatic activity only in presence of cations, optimal concentration 1.0 mM
Pseudomonas aeruginosa
2.7.7.46
Mg2+
shows enzymatic activity only in presence of cations, optimal concentration 2.0 mM
Pseudomonas aeruginosa
2.7.7.46
additional information
K+ and Na+ have almost no effect, NH4+ has a low enhancing effect
Pseudomonas aeruginosa
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.1.81
additional information
-
amino acid analysis
Pseudomonas aeruginosa
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2.7.7.46
ATP + gentamicin A
Pseudomonas aeruginosa
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
diphosphate + 2''-adenylylgentamicin A
-
Pseudomonas aeruginosa
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.1.81
plasmid pPK237 transferred to and expressed in E. coli K12, affinity chromatography
Pseudomonas aeruginosa
2.7.7.46
plasmid pPK237, purified from an E. coli K12 host RC85F-
Pseudomonas aeruginosa
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
2.3.1.81
34
-
-
Pseudomonas aeruginosa
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
2.3.1.81
-20C, partially purified, several months
Pseudomonas aeruginosa
2.3.1.81
-20C, purified, several days
Pseudomonas aeruginosa
2.7.7.46
4C, activity remains only for a few hours
Pseudomonas aeruginosa
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2.3.1.81
acetyl-CoA + dibekacin
acetylation at 8% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3''-acetyldibekacin
-
-
-
?
2.3.1.81
acetyl-CoA + gentamicin A
acetylation at 6% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin A
-
-
-
?
2.3.1.81
acetyl-CoA + gentamicin C
acetylation at 86% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin C
-
-
-
?
2.3.1.81
acetyl-CoA + gentamicin C1
best substrate
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin C1
-
-
-
?
2.3.1.81
acetyl-CoA + gentamicin C1a
acetylation at 60% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin C1a
-
637300
Pseudomonas aeruginosa
?
2.3.1.81
acetyl-CoA + gentamicin C2
acetylation at 84% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylgentamicin C2
-
-
-
?
2.3.1.81
acetyl-CoA + kanamycin A
acetylation at 7% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylkanamycin A
-
-
-
?
2.3.1.81
acetyl-CoA + sisomicin
acetylation at 51% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetylsisomicin
-
-
-
?
2.3.1.81
acetyl-CoA + tobramycin
acetylation at 6% the rate of gentamicin C1
637300
Pseudomonas aeruginosa
CoA + N3'-acetyltobramycin
-
-
-
?
2.3.1.81
additional information
no substrates are amikacin
637300
Pseudomonas aeruginosa
?
-
-
-
-
2.3.1.81
additional information
lividomycin A, butirosin A and B
637300
Pseudomonas aeruginosa
?
-
-
-
-
2.7.7.46
ATP + dibekacin
-
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylyldibekacin
-
-
-
r
2.7.7.46
ATP + gentamicin A
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylgentamicin A
-
637300
Pseudomonas aeruginosa
r
2.7.7.46
ATP + gentamicin C1
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylgentamicin C1
-
-
-
-
2.7.7.46
ATP + gentamicin C1a
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylgentamicin C1a
-
-
-
-
2.7.7.46
ATP + gentamicin C2
gentamicin A, C1, C1a, or C2 can act as acceptors, dATP, CTP, ITP or GTP can act as donors
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylgentamicin C2
-
-
-
-
2.7.7.46
ATP + kanamycin A
poor substrate
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylkanamycin A
-
-
-
r
2.7.7.46
ATP + sisomicin
-
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylylsisomicin
-
-
-
r
2.7.7.46
ATP + tobramycin
poor substrate
637300
Pseudomonas aeruginosa
diphosphate + 2''-adenylyltobramycin
-
-
-
r
2.7.7.46
additional information
amikacin, neomycin, lividomycin and butirosin A and B are no substrates
637300
Pseudomonas aeruginosa
?
-
-
-
-
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
2.3.1.81
37
-
assay at
Pseudomonas aeruginosa
Temperature Stability [C] (protein specific)
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
2.3.1.81
40
-
above, inactivation within 2 min
Pseudomonas aeruginosa
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.1.81
7
7.6
-
Pseudomonas aeruginosa
2.7.7.46
7.9
-
-
Pseudomonas aeruginosa