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Literature summary for 2.1.1.315 extracted from

  • Xu, J.; Mahmud, T.; Floss, H.G.
    Isolation and characterization of 27-O-demethylrifamycin SV methyltransferase provides new insights into the post-PKS modification steps during the biosynthesis of the antitubercular drug rifamycin B by Amycolatopsis mediterranei S699 (2003), Arch. Biochem. Biophys., 411, 277-288.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli as a polyhistidine-tagged polypeptide Amycolatopsis mediterranei

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ inhibits slightly Amycolatopsis mediterranei
Co2+ strong inhibition Amycolatopsis mediterranei
Mg2+ inhibits slightly Amycolatopsis mediterranei
Ni2+ strong inhibition Amycolatopsis mediterranei
Zn2+ strong inhibition Amycolatopsis mediterranei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.018
-
27-O-demethylrifamycin SV pH 8.0, 30°C Amycolatopsis mediterranei
0.0193
-
S-adenosyl-L-methionine pH 8.0, 30°C Amycolatopsis mediterranei

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the purified enzyme does not require a divalent cation for catalytic activity Amycolatopsis mediterranei

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
33800
-
x * 33800, His-tagged protein, calculated from sequence Amycolatopsis mediterranei
100000
-
non-denaturing PAGE Amycolatopsis mediterranei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-adenosyl-L-methionine + 27-O-demethylrifamycin SV Amycolatopsis mediterranei the enzyme is involved in biosynthesis of rifamycin B S-adenosyl-L-homocysteine + rifamycin SV
-
?
S-adenosyl-L-methionine + 27-O-demethylrifamycin SV Amycolatopsis mediterranei S699 the enzyme is involved in biosynthesis of rifamycin B S-adenosyl-L-homocysteine + rifamycin SV
-
?

Organism

Organism UniProt Comment Textmining
Amycolatopsis mediterranei
-
-
-
Amycolatopsis mediterranei S699
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information poor methylation of S-adenosyl-L-methionine Amycolatopsis mediterranei ?
-
?
additional information poor methylation of S-adenosyl-L-methionine Amycolatopsis mediterranei S699 ?
-
?
S-adenosyl-L-methionine + 27-O-demethylrifamycin SV the enzyme is involved in biosynthesis of rifamycin B Amycolatopsis mediterranei S-adenosyl-L-homocysteine + rifamycin SV
-
?
S-adenosyl-L-methionine + 27-O-demethylrifamycin SV the enzyme does not catalyze the methylation of rifamycin SV or rifamycin W Amycolatopsis mediterranei S-adenosyl-L-homocysteine + rifamycin SV
-
?
S-adenosyl-L-methionine + 27-O-demethylrifamycin SV the enzyme is involved in biosynthesis of rifamycin B Amycolatopsis mediterranei S699 S-adenosyl-L-homocysteine + rifamycin SV
-
?
S-adenosyl-L-methionine + 27-O-demethylrifamycin SV the enzyme does not catalyze the methylation of rifamycin SV or rifamycin W Amycolatopsis mediterranei S699 S-adenosyl-L-homocysteine + rifamycin SV
-
?

Subunits

Subunits Comment Organism
? x * 33800, His-tagged protein, calculated from sequence Amycolatopsis mediterranei

Synonyms

Synonyms Comment Organism
AdoMet:27-O-demethylrifamycin SV methyltransferase
-
Amycolatopsis mediterranei

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Amycolatopsis mediterranei

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
87
-
27-O-demethylrifamycin SV pH 8.0, 30°C Amycolatopsis mediterranei

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5 8
-
Amycolatopsis mediterranei
8
-
assay at Amycolatopsis mediterranei

pH Range

pH Minimum pH Maximum Comment Organism
6 8.5 pH 6.0: about 20% of maximal activity, pH 8.5: about 70% of maximal activity Amycolatopsis mediterranei

General Information

General Information Comment Organism
malfunction frameshift inactivation of rif orf14 generates a mutant that produces no rifamycin B, but accumulates 27-O-demethylrifamycin SV as the major new metabolite, together with a small quantity of 27-O-demethyl-25-O-desacetylrifamycin SV Amycolatopsis mediterranei

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4833
-
27-O-demethylrifamycin SV pH 8.0, 30°C Amycolatopsis mediterranei