Information on EC 1.14.13.B30 - rifampicin monooxygenase

for references in articles please use BRENDA:EC1.14.13.B30
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.13.B30
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
rifampicin monooxygenase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
rifampicin + NAD(P)H + O2 = 2'-N-hydroxyrifampicin + NAD(P)+ + H2O
show the reaction diagram
SYSTEMATIC NAME
IUBMB Comments
rifampicin:NAD(P)H:oxygen oxidoreductase (2'-N-hydroxyrifampicin-forming)
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
rifampicin + NAD(P)H + O2
2'-N-hydroxyrifampicin + NAD(P)+ + H2O
show the reaction diagram
rifampicin + NADH + O2
2'-N-hydroxy-4-oxo-rifampicin + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
rifampicin + NADPH + O2
2'-N-hydroxy-4-oxo-rifampicin + NADP+ + H2O
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
rifampicin + NADH + O2
2'-N-hydroxy-4-oxo-rifampicin + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
rifampicin + NADPH + O2
2'-N-hydroxy-4-oxo-rifampicin + NADP+ + H2O
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADH
-
RifMO exhibits only a 3fold coenzyme preference for NADPH over NADH
NADPH
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.266
NADH
-
pH 7.5, 25°C, recombinant enzyme, oxygen consumption measurement
0.075 - 0.09
NADPH
0.0045 - 0.006
rifampicin
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.3
NADH
-
pH 7.5, 25°C, recombinant enzyme, oxygen consumption measurement
1.3 - 4
NADPH
3.2 - 4
rifampicin
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
12.6
NADH
-
pH 7.5, 25°C, recombinant enzyme, oxygen consumption measurement
17.6 - 43.1
NADPH
222.4 - 701.6
rifampicin
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
51360
-
calculated from amino acid sequence
51890
calculated from amino acid sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 51400, detagged recombinant enzyme, SDS-PAGE
homodimer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
enzyme RIFMO complexed with rifampicin represents the precatalytic conformation that occurs before formation of the ternary E-RIF-NADPH complex, crystallization of the apoenzyme and the enzyme in complex with substrate rifamicin, hanging drop vapor diffusion method, mixing in a 1:1 ratio of 10.8 mg/ml protein in 25 mM HEPES and 100 mM NaCl, pH 7.5, and for the complex crystals 5 mM rifampicin, with reservoir solution containing 2.4 M sodium acetate trihydrate, pH 7.0, and PEG 3350 or PEG 4000 as precipitant, room temperature, microseeding, X-ray diffraction structure determination and analysis at 1.8-2.1 A resolution, small angle x-ray scattering, modeling
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His6-tagged enzyme from Escherichia coli to homogeneity by nickel affinity chromatography, cleavage of the His6-tag by thrombin
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3)
-
expressed in Escherichia coli MM294 and in Rhodococcus erythropolis Ri8, a nocardioform mutant of R. erythropolis ATCC 12674 selected on the basis of its inability to inactivate rifampin. The cloned gene refers resistance to rifamycin
gene rox, recombinant expression of His6-tagged enzyme in Escherichia coli
-
gene rox, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli
-