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Literature summary extracted from

  • Angelatou, F.; Litsas, S.B.; Kontomichalou, P.
    Purification and properties of two gentamicin-modifying enzymes, coded by a single plasmid pPK237 originating from Pseudomonas aeruginosa (1982), J. Antibiot., 35, 235-244.
    View publication on PubMed

Application

EC Number Application Comment 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 (Comment) 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 Comment 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 Comment Organism Structure
2.3.1.81 0.066
-
 gentamicin C1
-
 Pseudomonas aeruginosa

Metals/Ions

EC Number Metals/Ions Comment 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] Comment Organism
2.3.1.81 additional information
-
 amino acid analysis Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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
-
 r

Organism

EC Number Organism UniProt Comment 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 (Comment) 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 [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.1.81 34
-
-
 Pseudomonas aeruginosa

Storage Stability

EC Number Storage Stability Organism
2.3.1.81 -20°C, partially purified, several months Pseudomonas aeruginosa
2.3.1.81 -20°C, purified, several days Pseudomonas aeruginosa
2.7.7.46 4°C, activity remains only for a few hours Pseudomonas aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.81 acetyl-CoA + dibekacin acetylation at 8% the rate of gentamicin C1 Pseudomonas aeruginosa CoA + N3''-acetyldibekacin
-
 ?
2.3.1.81 acetyl-CoA + gentamicin A acetylation at 6% the rate of gentamicin C1 Pseudomonas aeruginosa CoA + N3'-acetylgentamicin A
-
 ?
2.3.1.81 acetyl-CoA + gentamicin C acetylation at 86% the rate of gentamicin C1 Pseudomonas aeruginosa CoA + N3'-acetylgentamicin C
-
 ?
2.3.1.81 acetyl-CoA + gentamicin C1 best substrate Pseudomonas aeruginosa CoA + N3'-acetylgentamicin C1
-
 ?
2.3.1.81 acetyl-CoA + gentamicin C1a acetylation at 60% the rate of gentamicin C1 Pseudomonas aeruginosa CoA + N3'-acetylgentamicin C1a
-
 ?
2.3.1.81 acetyl-CoA + gentamicin C2 acetylation at 84% the rate of gentamicin C1 Pseudomonas aeruginosa CoA + N3'-acetylgentamicin C2
-
 ?
2.3.1.81 acetyl-CoA + kanamycin A acetylation at 7% the rate of gentamicin C1 Pseudomonas aeruginosa CoA + N3'-acetylkanamycin A
-
 ?
2.3.1.81 acetyl-CoA + sisomicin acetylation at 51% the rate of gentamicin C1 Pseudomonas aeruginosa CoA + N3'-acetylsisomicin
-
 ?
2.3.1.81 acetyl-CoA + tobramycin acetylation at 6% the rate of gentamicin C1 Pseudomonas aeruginosa CoA + N3'-acetyltobramycin
-
 ?
2.3.1.81 additional information no substrates are amikacin Pseudomonas aeruginosa ?
-
 ?
2.3.1.81 additional information lividomycin A, butirosin A and B Pseudomonas aeruginosa ?
-
 ?
2.7.7.46 ATP + dibekacin
-
 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 Pseudomonas aeruginosa diphosphate + 2''-adenylylgentamicin A
-
 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 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 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 Pseudomonas aeruginosa diphosphate + 2''-adenylylgentamicin C2
-
 ?
2.7.7.46 ATP + kanamycin A poor substrate Pseudomonas aeruginosa diphosphate + 2''-adenylylkanamycin A
-
 r
2.7.7.46 ATP + sisomicin
-
 Pseudomonas aeruginosa diphosphate + 2''-adenylylsisomicin
-
 r
2.7.7.46 ATP + tobramycin poor substrate Pseudomonas aeruginosa diphosphate + 2''-adenylyltobramycin
-
 r
2.7.7.46 additional information amikacin, neomycin, lividomycin and butirosin A and B are no substrates Pseudomonas aeruginosa ?
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.81 37
-
 assay at Pseudomonas aeruginosa

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.3.1.81 40
-
 above, inactivation within 2 min Pseudomonas aeruginosa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.81 7 7.6
-
 Pseudomonas aeruginosa
2.7.7.46 7.9
-
-
 Pseudomonas aeruginosa