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Literature summary extracted from

  • Charavgi, M.D.; Dimarogona, M.; Topakas, E.; Christakopoulos, P.; Chrysina, E.D.
    The structure of a novel glucuronoyl esterase from Myceliophthora thermophila gives new insights into its role as a potential biocatalyst (2013), Acta Crystallogr. Sect. D, 69, 63-73 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.117 gene ge2, sequence comparisons, recombinant expression of His-tagged wild-type and mutant enzymes in Pichia pastoris Thermothelomyces thermophilus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.1.117 purified unliganded wild-type and mutant S213A enzymes, and S213A mutant in complex with a substrate analogue methyl 4-O-methyl-beta-D-glucopyranuronate, sitting drop vapour diffusion method, mixing of 10 mg/ml protein in 20 mM Tris-HCl, pH 8.0, with 30% w/v PEG 3350, 0.1 M Tris-HCl, pH 8.0, for the mutant and 0.1 M sodium acetate, pH 4.6, 25% v/v PEG 550 MME for the wild-type as precipitant solutions, at 16°C, X-ray diffraction structure determination and analysis at 1.55 A, 1.9 A, and 2.35 A resolution, respectively Thermothelomyces thermophilus
3.1.1.117 wild-type and mutant S213A, to 1.55 A and 1.9 A resolution, and mutant S213A in complex with substrate analogue, methyl 4-O-methyl-beta-D-glucopyranuronate. Serine-type hydrolase, its overall architecture follows the three-layer alphabetaalpha-sandwich hydrolase fold with a Rossmann-fold topology. Ser213, His346 and Glu236 are putative catalytic triad residues Thermothelomyces thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.117 S213A mutation in putative catalytic triad residue, crystallization data Thermothelomyces thermophilus
3.1.1.117 S213A site-directed mutagenesis, the S213A mutant exhibits a complete loss of enzyme activity towards methyl 4-O-methyl-D-glucopyranuronate, inactive mutant Thermothelomyces thermophilus

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.117 Thermothelomyces thermophilus G2QJR6 Sporotrichum thermophile
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3.1.1.117 Thermothelomyces thermophilus G2QJR6 member of Carbohydrate Esterase Family 15
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3.1.1.117 Thermothelomyces thermophilus ATCC 42464 G2QJR6 Sporotrichum thermophile
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3.1.1.117 Thermothelomyces thermophilus DSM 1799 G2QJR6 member of Carbohydrate Esterase Family 15
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.117 recombinant His-tagged wild-type and mutant enzymes from Pichia pastoris by nickel affinity chromatography Thermothelomyces thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.117 methyl 4-O-methyl-D-glucopyranuronate + H2O
-
 Thermothelomyces thermophilus methanol + 4-O-methyl-D-glucopyranuronate
-
 ?
3.1.1.117 methyl 4-O-methyl-D-glucopyranuronate + H2O
-
 Thermothelomyces thermophilus ATCC 42464 methanol + 4-O-methyl-D-glucopyranuronate
-
 ?
3.1.1.117 methyl 4-O-methyl-D-glucopyranuronate + H2O
-
 Thermothelomyces thermophilus DSM 1799 methanol + 4-O-methyl-D-glucopyranuronate
-
 ?
3.1.1.117 methyl-4-O-methyl-D-glucuronate + H2O
-
 Thermothelomyces thermophilus methanol + 4-O-methyl-D-glucuronate
-
 ?
3.1.1.117 methyl-4-O-methyl-D-glucuronate + H2O
-
 Thermothelomyces thermophilus ATCC 42464 methanol + 4-O-methyl-D-glucuronate
-
 ?
3.1.1.117 methyl-4-O-methyl-D-glucuronate + H2O
-
 Thermothelomyces thermophilus DSM 1799 methanol + 4-O-methyl-D-glucuronate
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.1.117 ? x * 43000, recombinant His-tagged enzyme, SDS-PAGE Thermothelomyces thermophilus
3.1.1.117 More the three-dimensional protein structures of wild-type and mutant enzymes have an alpha/beta-hydrolase fold with a three-layer alphabetaalpha-sandwich architecture and a Rossmann topology and comprise one molecule per asymmetric unit Thermothelomyces thermophilus

Synonyms

EC Number Synonyms Comment Organism
3.1.1.117 4-O-methyl-glucuronoyl methylesterase
-
 Thermothelomyces thermophilus
3.1.1.117 Ge2
-
 Thermothelomyces thermophilus
3.1.1.117 glucuronoyl esterase
-
 Thermothelomyces thermophilus
3.1.1.117 StGE2
-
 Thermothelomyces thermophilus

General Information

EC Number General Information Comment Organism
3.1.1.117 evolution the enzyme is a member of the CE15 family of carbohydrate esterases Thermothelomyces thermophilus
3.1.1.117 additional information three-dimensional homology structure modelling of StGE2 by molecular replacement using the structure of Cip2_GE (PDB ID 3pic, UniProt ID G0RV93) from Hypocrea jecorina strain QM6a as a starting model. The three-dimensional protein structures of wild-type and mutant enzymes have an alpha/beta-hydrolase fold with a three-layer alphabetaalpha-sandwich architecture and a Rossmann topology and comprise one molecule per asymmetric unit. The residues lining the H12 alpha-helix and the preceding loop region are distorted compared with those in the Cip2_GE structure since the N-linked glycosylation sequence motif (Asn-X-Ser) and the N-acetylglucosamine molecule bound at Asn447 observed in the latter are missing in StGE2 (Asn-X-Ala) is the corresponding motif in StGE2. Determination of the StGE2 catalytic site structure, overview. The catalytic triad residues, namely Ser213, Glu236 and His346, participate in a concrete ready-for-nucleophilic-attack configuration Thermothelomyces thermophilus