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Literature summary for 1.14.13.231 extracted from

  • 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 (2010), BMC Biochem., 11, 34.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
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 Bacteroides fragilis

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Bacteroides fragilis

Protein Variants

Protein Variants Comment Organism
D311A mutation in a highly conserved residue in the FAD-binding site, inactive Bacteroides fragilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.083
-
doxycycline presence of 0.0002% arabinose, pH not specified in the publication, temperature not specified in the publication Bacteroides fragilis
0.11
-
chlorotetracycline presence of 0.0002% arabinose, pH not specified in the publication, temperature not specified in the publication Bacteroides fragilis
0.13
-
anhydrotetracycline presence of 0.0002% arabinose, pH not specified in the publication, temperature not specified in the publication Bacteroides fragilis
0.16
-
anhydrotetracycline presence of 0.02% arabinose, pH not specified in the publication, temperature not specified in the publication Bacteroides fragilis

Organism

Organism UniProt Comment Textmining
Bacteroides fragilis
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
anhydrotetracycline + NADPH + H+ + O2
-
Bacteroides fragilis 11a-hydroxyanhydrotetracycline + NADP+ + H2O
-
?
chlorotetracycline + NADPH + H+ + O2
-
Bacteroides fragilis 11a-hydroxychlorotetracycline + NADP+ + H2O
-
?
doxycycline + NADPH + H+ + O2
-
Bacteroides fragilis 11a-hydroxydoxycycline + NADP+ + H2O
-
?

Synonyms

Synonyms Comment Organism
TetX
-
Bacteroides fragilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.3
-
chlorotetracycline presence of 0.0002% arabinose, pH not specified in the publication, temperature not specified in the publication Bacteroides fragilis
0.4
-
anhydrotetracycline presence of 0.02% arabinose, pH not specified in the publication, temperature not specified in the publication Bacteroides fragilis
0.6
-
doxycycline presence of 0.0002% arabinose, pH not specified in the publication, temperature not specified in the publication Bacteroides fragilis
0.7
-
anhydrotetracycline presence of 0.0002% arabinose, pH not specified in the publication, temperature not specified in the publication Bacteroides fragilis