Literature summary extracted from

  • Claxton, H.B.; Akey, D.L.; Silver, M.K.; Admiraal, S.J.; Smith, J.L.
    Structure and functional analysis of RifR, the type II thioesterase from the rifamycin biosynthetic pathway (2009), J. Biol. Chem., 284, 5021-5029.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.1.2.20 additional information movement of a helical lid controls access of substrates to the active site of RifR. The active site of RifR is a classic catalytic triad comprising residues Ser94, Asp200, and His228 Amycolatopsis mediterranei

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.2.20 expressed in Escherichia coli strain BL21 StarTM Amycolatopsis mediterranei

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.2.20 by hanging drop vapor diffusion at 4°C. Two RifR polypeptides in the asymmetric unit (P2(1): a=39.5 A, b=94.6 A, c=63.2 A, beta=90.55°) Amycolatopsis mediterranei

Engineering

EC Number Protein Variants Comment Organism
3.1.2.20 S94A thioesterase activity is effectively eliminated for all substrates except with isobutyryl-CoA and propionyl-CoA. No breakdown of [14C]acyl-S639A RifM1 substrates Amycolatopsis mediterranei

Organism

EC Number Organism UniProt Comment Textmining
3.1.2.20 Amycolatopsis mediterranei Q7BUF9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.2.20 by gel filtration and metal affinity column Amycolatopsis mediterranei

Storage Stability

EC Number Storage Stability Organism
3.1.2.20 -80°C, 50 mM HEPES buffer, pH 7.5, 50 mM NaCl, 1 mM EDTA, 1 mM tris-(2-carboxyethyl)phosphine, 10% (v/v) glycerol Amycolatopsis mediterranei

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.2.20 acetyl-CoA + H2O
-
Amycolatopsis mediterranei CoA + acetate
-
?
3.1.2.20 propionyl-CoA + H2O
-
Amycolatopsis mediterranei CoA + propionate
-
?
3.1.2.20 butyryl-CoA + H2O
-
Amycolatopsis mediterranei butanoate + CoA
-
?
3.1.2.20 octanoyl-CoA + H2O
-
Amycolatopsis mediterranei CoA + octanoate
-
?
3.1.2.20 decanoyl-CoA + H2O
-
Amycolatopsis mediterranei CoA + decanoate
-
?
3.1.2.20 hexanoyl-CoA + H2O
-
Amycolatopsis mediterranei CoA + hexanoate
-
?
3.1.2.20 malonyl-CoA + H2O
-
Amycolatopsis mediterranei CoA + malonate
-
?
3.1.2.20 isobutyryl-CoA + H2O
-
Amycolatopsis mediterranei CoA + isobutyrate
-
?
3.1.2.20 methylmalonyl-CoA + H2O
-
Amycolatopsis mediterranei CoA + methylmalonate
-
?
3.1.2.20 acetyl-RifM1 + H2O
-
Amycolatopsis mediterranei ?
-
?
3.1.2.20 methylmalonyl-RifM1 + H2O
-
Amycolatopsis mediterranei ?
-
?
3.1.2.20 propionyl-RifM1 + H2O
-
Amycolatopsis mediterranei ?
-
?

Subunits

EC Number Subunits Comment Organism
3.1.2.20 monomer in solution, gel filtration Amycolatopsis mediterranei

Synonyms

EC Number Synonyms Comment Organism
3.1.2.20 type II thioesterase
-
Amycolatopsis mediterranei
3.1.2.20 TEII
-
Amycolatopsis mediterranei
3.1.2.20 RifR
-
Amycolatopsis mediterranei