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Literature summary extracted from

  • Kim, C.; Hesek, D.; Zajicek, J.; Vakulenko, S.B.; Mobashery, S.
    Characterization of the bifunctional aminoglycoside-modifying enzyme ANT(3)-Ii/AAC(6)-IId from Serratia marcescens (2006), Biochemistry, 45, 8368-8377.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.47 cloned in Escherichia coli BL21 Serratia marcescens

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.1.82 6'-N-acetylkanamycin A competitive versus kanamycin A and noncompetitive/mixed versus acetyl-CoA Serratia marcescens
2.3.1.82 butyryl-CoA uncompetitive versus kanamycin A and competitive versus acetyl CoA Serratia marcescens
2.3.1.82 CoASH uncompetitive versus kanamycin A and noncompetitive/mixed versus acetyl-CoA Serratia marcescens
2.3.1.82 paromomycin competitive versus kanamycin A and noncompetitive/mixed versus acetyl-CoA Serratia marcescens
2.7.7.47 9-O-(adenosine-5'-phophoryl)spectinomycin product inhibition of ANT(3'')-Ii domain Serratia marcescens
2.7.7.47 AMP-CPP dead-end inhibition of ANT(3'')-Ii domain Serratia marcescens
2.7.7.47 kanamycin A dead-end inhibition of ANT(3'')-Ii domain Serratia marcescens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.82 0.012
-
 kanamycin A pH 7.5 Serratia marcescens
2.3.1.82 0.015
-
 gentamicin pH 7.5 Serratia marcescens
2.3.1.82 0.036
-
 netilmicin pH 7.5 Serratia marcescens
2.3.1.82 0.044
-
 acetyl-CoA pH 7.5 Serratia marcescens
2.3.1.82 0.313
-
 amikacin pH 7.5 Serratia marcescens
2.7.7.47 0.0013
-
 streptomycin kinetic parameter of the ANT(3'')-Ii domain of the enzyme Serratia marcescens
2.7.7.47 0.0014
-
 spectinomycin kinetic parameter of the ANT(3'')-Ii domain of the enzyme Serratia marcescens
2.7.7.47 0.014
-
 ATP kinetic parameter of the ANT(3'')-Ii domain of the enzyme Serratia marcescens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.47 Mg2+ assay with 15 mM MgCl2 Serratia marcescens

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.82 Serratia marcescens
-
-
-
2.7.7.47 Serratia marcescens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.47 DEAE anion-exchange column and gentamycin affinity column Serratia marcescens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.82 acetyl-CoA + amikacin
-
 Serratia marcescens CoA + N6'-acetylamikacin
-
 ?
2.3.1.82 acetyl-CoA + gentamicin
-
 Serratia marcescens CoA + N6'-acetylgentamicin
-
 ?
2.3.1.82 acetyl-CoA + kanamycin A the bifunctional antibiotic resistance enzyme from Serratia marcescens catalyzes adenylation and acetylation of aminoglycoside antibiotics. The structure assignment of the enzymic products indicated that acetylation takes place on the 6'-amine of kanamycin A and the adenylation on 3''- and 9-hydroxyl groups of streptomycin and spectinomycin, respectively. The adenyltransferase domain appears to be highly specific to spectinomycin and streptomycin, while the acetyltransferase domain shows a broad substrate profile. Initial velocity patterns indicate that both domains follow a sequential kinetic mechanism Serratia marcescens CoA + N6'-acetylkanamycin A
-
 ?
2.3.1.82 acetyl-CoA + netilmicin
-
 Serratia marcescens CoA + N6'-acetylnetilmicin
-
 ?
2.7.7.47 ATP + spectinomycin
-
 Serratia marcescens diphosphate + 9-adenylylspectinomycin
-
 ?
2.7.7.47 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 Serratia marcescens diphosphate + 3''-adenylylstreptomycin
-
 ?
2.7.7.47 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 Serratia marcescens ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.1.82 ANT(3'')-Ii/AAC(6')-IId bifunctional enzyme catalyzes adenylation and acetylation of aminoglycoside antibiotics Serratia marcescens
2.7.7.47 ANT(3'')-Ii/AAC(6')-IId
-
 Serratia marcescens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.47 additional information
-
 assay is performed at room temperature Serratia marcescens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.82 0.05
-
 amikacin pH 7.5 Serratia marcescens
2.3.1.82 0.2
-
 netilmicin pH 7.5 Serratia marcescens
2.3.1.82 0.5
-
 gentamicin pH 7.5 Serratia marcescens
2.3.1.82 0.8
-
 acetyl-CoA pH 7.5 Serratia marcescens
2.3.1.82 0.8
-
 kanamycin A pH 7.5 Serratia marcescens
2.7.7.47 0.5
-
 ATP turnover number of ANT(3'')-Ii domain Serratia marcescens
2.7.7.47 0.5
-
 spectinomycin turnover number of ANT(3'')-Ii domain Serratia marcescens
2.7.7.47 0.6
-
 streptomycin turnover number of ANT(3'')-Ii domain Serratia marcescens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.47 7.5
-
 assay at Serratia marcescens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.3.1.82 0.025
-
 butyryl-CoA versus acetyl-CoA, K(is) Serratia marcescens
2.3.1.82 0.04
-
 6'-N-acetylkanamycin A versus kanamycin A, K(is) Serratia marcescens
2.3.1.82 0.042
-
 CoASH versus acetyl-CoA, K(is) Serratia marcescens
2.3.1.82 0.043
-
 CoASH versus acetyl-CoA, K(ii) Serratia marcescens
2.3.1.82 0.06
-
 paromomycin versus kanamycin A, K(is) Serratia marcescens
2.3.1.82 0.069
-
 CoASH versus kanamycin A, K(ii) Serratia marcescens
2.3.1.82 0.117
-
 paromomycin versus acetyl-CoA, K(is) Serratia marcescens
2.3.1.82 0.145
-
 paromomycin versus acetyl-CoA, K(ii) Serratia marcescens
2.3.1.82 0.15
-
 6'-N-acetylkanamycin A versus acetyl-CoA, K(is) Serratia marcescens
2.3.1.82 0.188
-
 butyryl-CoA versus kanamycin A, K(ii) Serratia marcescens
2.3.1.82 0.385
-
 6'-N-acetylkanamycin A versus acetyl-CoA, K(ii) Serratia marcescens
2.7.7.47 0.004
-
 kanamycin A dead-end inhibition of ANT(3'')-Ii domain, substrate 0.02 mM spectinomycin, competitive inhibition Serratia marcescens
2.7.7.47 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
2.7.7.47 0.017
-
 AMP-CPP dead-end inhibition of ANT(3'')-Ii domain, substrate 0.1 mM ATP, competitive inhibition Serratia marcescens
2.7.7.47 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
2.7.7.47 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
2.7.7.47 0.093
-
 9-O-(adenosine-5'-phophoryl)spectinomycin product inhibition of ANT(3'')-Ii domain, substrate 0.1 mM ATP, competitive inhibition Serratia marcescens