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Information on EC 1.14.13.231 - tetracycline 11a-monooxygenase and Organism(s) Bacteroides fragilis and UniProt Accession Q01911

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IUBMB Comments
A flavoprotein (FAD). This bacterial enzyme confers resistance to all clinically relevant tetracyclines when expressed under aerobic conditions. The hydroxylated products are very unstable and lead to intramolecular cyclization and non-enzymic breakdown to undefined products.
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Bacteroides fragilis
UNIPROT: Q01911
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The taxonomic range for the selected organisms is: Bacteroides fragilis
The enzyme appears in selected viruses and cellular organisms
Synonyms
tetx2, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetracycline resistance protein from transposon Tn4351/Tn4400
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
RH + [reduced NADPH-hemoprotein reductase] + O2 = ROH + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
tetracycline substrates, initial monohydroxylation at position 11a followed by intramolecular cyclization and non-enzymatic breakdown to undefined products
-
SYSTEMATIC NAME
IUBMB Comments
tetracycline,NADPH:oxygen oxidoreductase (11a-hydroxylating)
A flavoprotein (FAD). This bacterial enzyme confers resistance to all clinically relevant tetracyclines when expressed under aerobic conditions. The hydroxylated products are very unstable and lead to intramolecular cyclization and non-enzymic breakdown to undefined products.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
tetracycline + NADPH + H+ + O2
11a-hydroxytetracycline + NADP+ + H2O
show the reaction diagram
-
-
-
?
anhydrotetracycline + NADPH + H+ + O2
11a-hydroxyanhydrotetracycline + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
chlorotetracycline + NADPH + H+ + O2
11a-hydroxychlorotetracycline + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
chlortetracycline + reduced flavoprotein + O2
11a-hydroxy-chlortetracycline + oxidized flavoprotein + H2O
show the reaction diagram
-
-
-
-
?
demeclocycline + reduced flavoprotein + O2
11a-hydroxy-demeclocycline + oxidized flavoprotein + H2O
show the reaction diagram
-
-
-
-
?
doxycycline + NADPH + H+ + O2
11a-hydroxydoxycycline + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
doxycycline + reduced flavoprotein + O2
11a-hydroxy-doxycycline + oxidized flavoprotein + H2O
show the reaction diagram
-
-
-
-
?
minocycline + reduced flavoprotein + O2
11a-hydroxy-minocycline + oxidized flavoprotein + H2O
show the reaction diagram
-
-
-
-
?
oxytetracycline + reduced flavoprotein + NADPH + O2
11a-hydroxy-oxytetracycline + oxidized flavoprotein + H2O + NADP+
show the reaction diagram
-
-
-
-
?
tetracycline + reduced flavoprotein + O2
11a-hydroxy-tetracycline + oxidized flavoprotein + H2O
show the reaction diagram
-
-
-
-
?
tigecycline + NADPH + H+ + O2
11a-hydroxytigecycline + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
tetracycline substrates, initial monohydroxylation at position 11a followed by intramolecular cyclization and non-enzymatic breakdown to undefined products
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.13 - 0.16
anhydrotetracycline
0.11
chlorotetracycline
-
presence of 0.0002% arabinose, pH not specified in the publication, temperature not specified in the publication
0.11
chlortetracycline
-
pH 8.5
0.0199
demeclocycline
-
pH 8.5
0.083 - 0.0837
doxycycline
0.0284
minocycline
-
pH 8.5
0.133
NADPH
-
pH 8.5
0.076
oxytetracycline
-
pH 8.5
0.054
tetracycline
-
pH 8.5
0.044
tigecycline
-
pH 8.5, temperature not specified in the publication
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.4 - 0.7
anhydrotetracycline
0.3
chlorotetracycline
-
presence of 0.0002% arabinose, pH not specified in the publication, temperature not specified in the publication
0.3
chlortetracycline
-
pH 8.5
0.2
demeclocycline
-
pH 8.5
0.6 - 0.63
doxycycline
0.12
minocycline
-
pH 8.5
1.11
NADPH
-
pH 8.5
1.3
oxytetracycline
-
pH 8.5
0.32
tetracycline
-
pH 8.5
0.36
tigecycline
-
pH 8.5, temperature not specified in the publication
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
8.3
tigecycline
-
pH 8.5, temperature not specified in the publication
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
-
tigecycline is a substrate for TetX and bacterial strains containing the TetX gene are resistant to tigecycline due to the modification of tigecycline by TetX to form 11a-hydroxytigecycline, which has a weakened ability to inhibit protein translation compared with tigecycline. 11a-Hydroxytigecycline forms a weaker complex with magnesium than tigecycline, magnesium coordination is critical for binding tetracycline to the ribosome
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
TETX_BACFG
388
0
43725
Swiss-Prot
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
43700
x * 43700, calculated
44000
-
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 43700, calculated
monomer
-
x * 44000, SDS-PAGE and deduced from gene sequence
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D311A
-
mutation in a highly conserved residue in the FAD-binding site, inactive
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant protein, purified protein gives a yellow solution
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
construction of a sensitive fluorescent reporter system to detect and characterize the activity of enzymes that act upon the antibiotic, tetracycline and its derivatives. In this system, expression of the lux operon is regulated by the tetracycline repressor, TetR, which is expressed from the same plasmid under the control of an arabinose-inducible promoter. Addition of very low concentrations of tetracycline derivatives, well below growth inhibitory concentrations, result in luminescence production. Introduction of a plasmid expressing TetX, a tetracycline-inactivating enzyme, causes a marked loss in luminescence due to enzyme-mediated reduction in the intracellular tetracycline concentration
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Yang, W.; Moore, I.F.; Koteva, K.P.; Bareich, D.C.; Hughes, D.W.; Wright, G.D.
TetX is a flavin-dependent monooxygenase conferring resistance to tetracycline antibiotics
J. Biol. Chem.
279
52346-52352
2004
Bacteroides fragilis, Bacteroides thetaiotaomicron (Q93L51)
Manually annotated by BRENDA team
Moore, I.F.; Hughes, D.W.; Wright, G.D.
Tigecycline is modified by the flavin-dependent monooxygenase TetX
Biochemistry
44
11829-11835
2005
Bacteroides fragilis
Manually annotated by BRENDA team
Yu, Z.; Reichheld, S.E.; Cuthbertson, L.; Nodwell, J.R.; Davidson, A.R.
Characterization of tetracycline modifying enzymes using a sensitive in vivo reporter system
BMC Biochem.
11
34
2010
Bacteroides fragilis
Manually annotated by BRENDA team
Speer, B.S.; Bedzyk, L.; Salyers, A.A.
Evidence that a novel tetracycline resistance gene found on two Bacteroides transposons encodes an NADP-requiring oxidoreductase
J. Bacteriol.
173
176-183
1991
Bacteroides fragilis (Q01911)
Manually annotated by BRENDA team