Information on EC 2.3.1.81 - aminoglycoside 3-N-acetyltransferase

-

-

?

4-bromo-thiophene-2-carbonyl-S-CoA + gentamicin

?

-

-

?

4-bromo-thiophene-2-carbonyl-S-CoA + neomycin B

?

-

-

?

4-bromo-thiophene-2-carbonyl-S-CoA + paromomycin

?

-

-

?

4-bromo-thiophene-2-carbonyl-S-CoA + tobramycin

?

-

-

?

5-bromo-thiophene-2-carbonyl-S-CoA + kanamycin A

?

-

-

?

5-bromo-thiophene-2-carbonyl-S-CoA + neomycin B

?

-

-

?

6-fluoropicolinyl-CoA + neomycin B

?

-

-

?

acetyl-CoA + a 2-deoxystreptamine antibiotic

CoA + N3-acetyl-2-deoxystreptamine antibiotic

acetyl-CoA + amikacin

CoA + N3'-acetyl-amikacin

-

-

?

acetyl-CoA + amikacin

CoA + N3'-acetylamikacin

acetyl-CoA + aminoglycoside-aminocyclitol

CoA + ?

Q4R0X5

-

675934

-

?

acetyl-CoA + apramycin

CoA + N3'-acetylapramycin

acetyl-CoA + butirosin

CoA + N3'-acetylbutirosin

-

-

?

acetyl-CoA + dactimicin

CoA + N4-acetyldactamicin + N4,N6'-diacetyldactimicin

-

the percentage of dactimicin affinity relative to gentamicin C1a is very high (74.6%) only with AAC(3)-I, to the most other enzymes tested, in spite of the bifunctional AAC(6')/APH(2''), dactamicin is stable

637305

-

?

acetyl-CoA + dibekacin

CoA + N3''-acetyldibekacin

acetyl-CoA + dibekacin

CoA + N3-acetyldibekacin

acetyl-CoA + fortimicin

CoA + N3-acetylfortimicin

-

-

?

acetyl-CoA + gentamicin

CoA + N3'-acetyl-gentamicin

-

-

?

acetyl-CoA + gentamicin

CoA + N3'-acetylgentamicin

acetyl-CoA + gentamicin A

CoA + N3'-acetylgentamicin A

-

acetylation at 6% the rate of gentamicin C1

-

?

acetyl-CoA + gentamicin C

CoA + N3'-acetylgentamicin C

-

acetylation at 86% the rate of gentamicin C1

-

?

acetyl-CoA + gentamicin C1

CoA + N3'-acetylgentamicin C1

acetyl-CoA + gentamicin C1a

CoA + N3'-acetylgentamicin C1a

acetyl-CoA + gentamicin C1A

CoA + N3-acetylgentamicin C1A

-

-

?

acetyl-CoA + gentamicin C2

CoA + N3'-acetylgentamicin C2

acetyl-CoA + isepamycin

CoA + N3'-acetylisepamycin

wild-type and recombinant enzyme

-

?

acetyl-CoA + kanamycin

CoA + N3'-acetylkanamycin

acetyl-CoA + kanamycin A

CoA + N3'-acetylkanamycin A

acetyl-CoA + kanamycin B

CoA + N3'-acetylkanamycin B

acetyl-CoA + kanamycin B

CoA + N3-acetylkanamycin B

acetyl-CoA + lividomycin A

CoA + ?

-

-

?

acetyl-CoA + neomycin

CoA + ?

-

-

?

acetyl-CoA + neomycin

CoA + N2'-acetylneomycin

wild-type and recombinant enzyme

-

?

acetyl-CoA + neomycin

CoA + N3-acetylneomycin

-

-

?

acetyl-CoA + neomycin B

CoA + ?

acetyl-CoA + neomycin B

CoA + N3-acetyl-neomycin B

acetyl-CoA + neomycin B

CoA + N3-acetylneomycin B

-

-

?

acetyl-CoA + netilmicin

CoA + N3'-acetylnetilmicin

acetyl-CoA + paromomycin

CoA + N3'-acetyl-paromomycin

acetyl-CoA + paromomycin

CoA + N3'-acetylparomomycin

-

-

?

acetyl-CoA + paromomycin

CoA + N3'-acetylparomomycin B

-

not E. coli

637299

-

?

acetyl-CoA + paromomycin

CoA + N3-acetylparamomycin

-

-

?

acetyl-CoA + ribostamycin

CoA + ?

acetyl-CoA + ribostamycin

CoA + N3-acetylribostamycin

-

-

?

acetyl-CoA + sisomicin

CoA + N3'-acetyl-sisomicin

acetyl-CoA + sisomicin

CoA + N3'-acetylsisomicin

acetyl-CoA + sisomicin

CoA + N3-acetylsisomicin

-

-

?

acetyl-CoA + sisomycin

CoA + N3'-acetylsisomycin

acetyl-CoA + spectinomycin

CoA + N3'-acetylspectinomycin

wild-type and recombinant enzyme

-

?

acetyl-CoA + streptomycin

CoA + ?

wild-type and recombinant enzyme

-

?

acetyl-CoA + tobramycin

CoA + N3'-acetyl-tobramycin

-

-

?

acetyl-CoA + tobramycin

CoA + N3'-acetyltobramycin

acetyl-CoA + tobramycin

CoA + N3-acetyltobramycin

-

-

?

butyryl-CoA + kanamycin A

CoA + N3'-butyryl-kanamycin A

-

-

?

butyryl-CoA + ribostamycin

CoA + ?

-

-

?

desulfo-CoA + ribostamycin

?

-

-

?

glutaryl-CoA + kanamycin A

?

-

-

?

glycyl-CoA + amikacin

CoA + N3'-glycyl-amikacin

-

-

?

glycyl-CoA + gentamicin

CoA + N3'-glycyl-gentamicin

-

-

?

glycyl-CoA + kanamycin A

CoA + N3'-glycyl-kanamycin A

-

-

?

glycyl-CoA + neomycin B

CoA + N3-glycylneomycin B

-

-

?

glycyl-CoA + sisomicin

CoA + N3'-glycyl-sisomicin

-

-

?

malonyl-CoA + amikacin

CoA + N3'-malonyl-amikacin

-

-

?

malonyl-CoA + gentamicin

CoA + N3'-malonyl-gentamicin

-

-

?

malonyl-CoA + kanamycin A

CoA + N3'-malonylkanamycin A

-

-

?

malonyl-CoA + neomycin B

CoA + N3-malonylneomycin B

-

-

?

malonyl-CoA + ribostamycin

CoA + ?

-

-

?

methylmalonyl-CoA + neomycin B

CoA + N3-methylmalonylneomycin B

-

-

?

N-propionyl-CoA + amikacin

CoA + N3'-propionyl-amikacin

-

-

?

N-propionyl-CoA + gentamicin

CoA + N3'-propionyl-gentamicin

-

-

?

n-propionyl-CoA + kanamycin A

CoA + N3'-n-propionylkanamycin A

-

-

?

N-propionyl-CoA + kanamycin A

CoA + N3'-propionyl-kanamycin A

-

-

?

N-propionyl-CoA + neomycin B

CoA + N3-propionylneomycin B

-

-

?

N-propionyl-CoA + paromomycin

CoA + N3'-propionyl-paromomycin

-

-

?

N-propionyl-CoA + sisomicin

CoA + N3'-propionyl-sisomicin

-

-

?

N-propionyl-CoA + tobramycin

CoA + N3'-propionyl-tobramycin

-

-

?

propionyl-CoA + ribostamycin

CoA + ?

-

-

?

additional information

?

-

17 entries

acetyl-CoA + a 2-deoxystreptamine antibiotic

CoA + N3-acetyl-2-deoxystreptamine antibiotic

acetyl-CoA + dibekacin

CoA + N3-acetyldibekacin

acetyl-CoA + kanamycin B

CoA + N3-acetylkanamycin B

additional information

?

-

acetyl-CoA

-

Ca2+

-

1.0 mM, about 100% activity enhancement

Mg2+

-

1.5 mM, about 100% activity enhancement

NH4+

-

slight activation

3-(dimethylamino)propylamine

-

0.5 mM, 30% residual activity

acetyltobramycin

-

amikacin

-

butyryl-CoA

-

dead-end inhibition pattern

CoA

-

product inhibition

desulfo-CoA

-

EDTA

-

above 0.5 mM

gentamicin C1a

-

substrate inhibition

kanamycin A

-

substrate inhibition

661049, 718870

kanamycin B

-

substrate inhibition

N-(2-naphthyl)-N-methyl-2,3-dioxo-1,2,3,4-tetrahydro-6-quinoxalinesulfonamide

lacks anti-Mycobacterium tuberculosis activity in the absence of kanamycin in either tested strain

-

N-(3-methoxyphenyl)-N-methyl-2,3-dioxo-1,2,3,4-tetrahydro-6-quinoxalinesulfonamide

inhibitor completely overcomes kanamycin resistance in Mycobacterium tuberculosis K204, lacks anti-Mycobacterium tuberculosis activity in the absence of kanamycin in either tested strain

N-cyclohexyl-3-(3-dimethylamino-propylamino)-propionamide

-

0.5 mM, 77% inhibition

672534

N-[2-(3,4-dimethoxyphenyl)-ethyl]-3-(3-dimethylamino-propylamino)-propionamide

-

0.5 mM, 63% inhibition

672534

N3-Acetyltobramycin

-

product inhibition

neomycin

-

netilmicin

-

ribostamycin

tobramycin

-

substrate inhibition

661049, 718870

Zn2+

-

additional information

-

0.0013 - 0.17

acetyl-CoA

8 entries

0.00096 - 11.07

amikacin

0.00067

apramycin

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

0.237

butirosin

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

1.16

butyryl-CoA

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

0.0092

dibekacin

0.0029 - 0.0189

gentamicin

0.002 - 0.066

gentamicin C1

0.00012

gentamicin C1a

-

0.003

gentamicin C2

-

pH 7.6, 25Â°C

0.00097

gentamycin

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

0.002 - 0.238

kanamycin A

0.0026 - 0.0802

kanamycin B

0.0036

lividomycin A

-

pH 7.6, 25Â°C

0.234 - 0.477

malonyl-CoA

0.0886

n-propionyl-CoA

-

cosubstrate kanamycin A, pH 7.6, 25Â°C

0.00036

neomycin

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

0.0012 - 0.0148

neomycin B

0.042

netilmicin

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

0.00015 - 0.0033

paromomycin

0.499

propionyl-CoA

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

0.0023 - 0.0031

ribostamycin

0.00057 - 0.015

sisomicin

0.00033 - 0.0015

sisomycin

0.0013 - 0.106

tobramycin

7 entries

additional information

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0.24 - 39.4

acetyl-CoA

0.124 - 0.5

amikacin

61

apramycin

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

3.5

butirosin

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

0.3

butyryl-CoA

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

0.17

dibekacin

0.041 - 0.182

gentamicin

9.7

gentamicin C1

-

pH 7.6, 25Â°C

18.4

gentamicin C2

-

pH 7.6, 25Â°C

40

gentamycin

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

0.012 - 80

kanamycin A

0.23 - 55

kanamycin B

33.8

lividomycin A

-

pH 7.6, 25Â°C

5.7 - 9

malonyl-CoA

10.2

n-propionyl-CoA

-

cosubstrate kanamycin A, pH 7.6, 25Â°C

10 - 60

neomycin

0.076 - 9.5

neomycin B

34

neomycin E

pH 7.6, 25Â°C

32

netilmicin

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

15 - 33.6

paromomycin

33

propionyl-CoA

in 50 mM HEPES, pH 7.5 and 0.1 mM 4,4'-dithiodipyridine, at pH 7.8 and 25Â°C

23.5 - 146

ribostamycin

0.121 - 24.5

sisomicin

46 - 51

sisomycin

0.089 - 162

tobramycin

89 - 2600

acetyl-CoA

18

dibekacin

4700

gentamicin C1

-

pH 7.6, 25Â°C

6100

gentamicin C2

-

pH 7.6, 25Â°C

10600 - 11200

kanamycin A

17 - 34400

kanamycin B

9500

lividomycin A

-

pH 7.6, 25Â°C

30

malonyl-CoA

-

cosubstrate kanamycin A, pH 7.6, 25Â°C

120

n-propionyl-CoA

-

cosubstrate kanamycin A, pH 7.6, 25Â°C

9100

neomycin B

-

pH 7.6, 25Â°C

10800

paromomycin

-

pH 7.6, 25Â°C

10600

ribostamycin

-

pH 7.6, 25Â°C

1600

sisomicin

-

pH 7.6, 25Â°C

30700 - 33400

sisomycin

400 - 60500

tobramycin

0.381

kanamycin A

-

pH 7.6, 25Â°C

0.0059

ribostamycin

-

pH 7.6, 25Â°C

0.00000024

N-(2-naphthyl)-N-methyl-2,3-dioxo-1,2,3,4-tetrahydro-6-quinoxalinesulfonamide

Mycobacterium tuberculosis

pH 8.0, 25Â°C

-

0.058

N-(3-methoxyphenyl)-N-methyl-2,3-dioxo-1,2,3,4-tetrahydro-6-quinoxalinesulfonamide

Mycobacterium tuberculosis

pH 8.0, 25Â°C

34

-

9.743

-

pH 7.2, 34Â°C

additional information

-

Vmax 0.102 (sisomicin), pH 7.2, 34Â°C

6

assay at

6.6

-

assay at

7 - 7.6

-

8

corresponds to the plateau region of the pH profile, where the velocity is maximal and the variations in velocity due to changes in pH are smallest

25

-

assay at

30

37

5.7

calculated from amino acid sequence

gene aacC2; diverse clinical isolates, gene aacC2

736260

UniProt

Arizona sp.

-

487549

-

Corynebacterium striatum BM4687

-

-

Escherichia coli BL21(DE3) pLysS

-

TrEMBL

Escherichia coli C600

containing R-factor JR88

Escherichia coli R135/C600

-

Escherichia coli JR88

-

637304

-

-

637299

-

-

672534

-

Klebsiella pneumoniae

-

487549

-

-

Mycobacterium tuberculosis H37Rv

UniProt

-

Mycobacteroides abscessus

UniProt

-

756710

B1MKV6

UniProt

Mycobacteroides abscessus DSM 44196

-

756710

B1MKV6

UniProt

Pseudomonas aeruginosa Travers PST

-

637299

-

Saccharophagus degradans

formerly Microbulbifer degradans

671371

-

Salmonella enterica

serovar Haifa, isolated from feces

703467

-

Salmonella enterica subsp. enterica serovar Kentucky

-

SwissProt

-

-

strain ATCC 21294

675934

Q4R0X5

TrEMBL

additional information

analysis of enzyme expression in several clinical isolates, overview

evolution

physiological function

additional information

Bacillus anthracis str. Ames

presence of genes encoding the following aminoglycoside-modifying enÂzymes: aph (3')-V1, aph (3')-Ia, aac (3)-Ia, aac (3) IIa, aac (6')-Ih, aac (6')-Ib and ant (2')-Ia responsible for resisÂtance in 4 different groups of clinical isolates, overview

7lao, download, 3D-view

7lap, download, 3D-view

18630

-

18649 Da predicted from nucleotide sequence, 18630 Da found by mass spectrometry

487550

28000

2 * 28000, enzyme in solution

39000

-

49000

SDS-PAGE

54000

dynamic light scattering

55750

calculated from amino acid sequence

60000

Sephacryl-S200 gel filtration

additional information

-

amino acid analysis

dimer

additional information

-

AAC-VIa displays a monomer-dimer equilibrium in the absence of substrates.The dimer formation is concentration-dependent between 5 microM and 100 microM enzyme with more dimeric form appearing at higher concentrations. Binding of ligands shifts the enzyme to monomeric state

side-chain modification

Serratia marcescens

-

cleavage of the hexahistidine tag by tobacco etch virus protease

487550

in complex with inhibitor N-(3-methoxyphenyl)-N-methyl-2,3-dioxo-1,2,3,4-tetrahydro-6-quinoxalinesulfonamide. Inhibitor binds Eis in the AG-binding pocket held by a conformationally malleable region of Eis (residues 28-37) bearing key hydrophobic residues

Mycobacteroides abscessus

structures of apo-enzyme and in complex with acetyl-CoA

B1MKV6

756710

in the apo form, in binary drug (sisomicin, neomycin, and paromomycin) and coenzyme A complexes, and in the ternary neomycin-CoASH complex

757396

X-ray structure of a recombinant enzyme, expressed in its selenomethionine substituted form, cocrystallization with 5 mM CoA at 4Â°C and pH: 7.8, crystal cryoprotection achieved with 17% glycerol and 8% 2R,3R-butanediol, active center found by X-ray analysis and model building

Serratia marcescens

-

487550

C204A

40

-

above, inactivation within 2 min

dialysis against EDTA with subsequent Mg2+-addition to dialyzed enzyme has no effect on activity

-

enzyme activity is significantly stabilized when preparations contain substrate

-

gel filtration leads to complete inactivation

-

-20Â°C, partially purified, several months

-

-20Â°C, purified, several days

-

-

Fast Flow Q-Sepharose anion exchange column chromatography and Phenyl Sepharose column chromatography

Ni2+-affinity chromatography

partial

-

plasmid pPK237 transferred to and expressed in E. coli K12, affinity chromatography

-

recombinant His-tagged enzyme from Escherichia coli strain BL21 Star(DE3) by nickel affinity chromatography and ultrafiltration

expressed in Escherichia coli

expressed in the pET-23a(+) expression vector

expression in Escherichia coli

expression in Escherichia coli DH5alpha

expression in Escherichia coli K802N by mating

expression of C- or N-terminally His-tagged enzyme from vectors pET22b, and pET28a or Int-pET19b-pps, respectively

-

gene aac(3)-Id, in a class 1 integron

Salmonella enterica

-

703467

gene aacC2, genotyping

736260

gne aac(3)-XI, DNA and amino acid sequence determination and analysis, genomic organization, the aac(3)-XI gene is located downstream from a gene for a transposase (TnpCx) and upstream from that for a two-component system sensor kinase, the sequence upstream from aac(3)-XI does not reveal any putative promoter, recombinant expression of His-tagged enzyme in Escherichia coli strains TOP10 and BL21 Star(DE3)

plasmid transfer pIJ702 to Streptomyces lividans TK21

Streptomyces griseus

-

487551

Pseudomonas aeruginosa plasmid pPK237 transferred to and expressed in Escherichia coli K12

-

wild-type and truncated enzymes expressed in Escherichia coli

-

medicine

synthesis

-

development of a methodology that utilizes aminoglycoside acetyltransferase and unnatural acyl-CoA analogues for the chemoenzymatic generation of regioselectively N-acylatedaminoglycoside analogues. Aminoglycosides are broad-spectrum antibiotics commonly used for the treatment of serious bacterial infections