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Literature summary for 3.1.1.117 extracted from

  • Ernst, H.A.; Mosbech, C.; Langkilde, A.E.; Westh, P.; Meyer, A.S.; Agger, J.W.; Larsen, S.
    The structural basis of fungal glucuronoyl esterase activity on natural substrates (2020), Nat. Commun., 11, 1026 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.1.117

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes in Pichia pastoris strain X-33 Cerrena unicolor

Crystallization (Commentary)

Crystallization (Comment) Organism
purified isolated deglycosylated wild-type and mutant S207A catalytic domains, X-ray diffraction structure determination and analysis at 1.46-1.96 A resolution, structure modelling by molecular replacement using Cip2 (PDB ID 3PIC) as search model Cerrena unicolor

Protein Variants

Protein Variants Comment Organism
additional information construction of a truncated mutant (DELTAWT), comprising residues 78-458 (corresponding to the catalytic domain only) and lacking the carbohydrate-binding domain (CBM1). Neither the truncation nor the deglycosylation affects protein stability significantly Cerrena unicolor
S270A site-directed mutagenesis, truncated mutant S270A is an inactive variant of the truncated mutant DELTAWT where the catalytic serine is replaced with an alanine. The inactive truncated variant S270A is destabilized by ca 3°C compared to variants with an intact catalytic triad, implying that some stabilizing interactions have been lost, but the overall conclusion is that CuGE is a highly stable enzyme with Ti's in the range of 72.2-75.8°C for all variants. Binding between truncated S270A and the 4-O-methyl-glucuronoyl moiety alone has about 10times higher dissociation constant compared to binding of aldotetrauronic acid (Um4XX-OH) Cerrena unicolor

Organism

Organism UniProt Comment Textmining
Cerrena unicolor A0A0A7EQR3 Daedalea unicolor
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Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein heterogenous N- and O-glycosylation. Deglycosylation by Endo H and alpha-1-2,3,6 mannosidase Cerrena unicolor

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His6-tagged wild-type and mutant enzymes from Pichia pastoris strain X-33 by nickel affinity chromatography Cerrena unicolor

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
methyl-4-O-methyl-D-glucuronate + H2O
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 Cerrena unicolor methanol + 4-O-methyl-D-glucuronate
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 ?
additional information analysis of the interactions between CuGE and the alpha-1,2-linked 4-O-methyl-D-glucuronoyl moieties on xylo-oligomers, the binding of the 4-O-methyl-alpha-D-glucuronoyl moiety is not influenced by the nature of the attached xylo-oligosaccharide. Enzyme-substrate binding analysis, overview Cerrena unicolor ?
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Subunits

Subunits Comment Organism
More the enzyme is modular, comprised of a catalytic and a carbohydrate-binding domain. CuGE is an elongated rigid molecule where the two domains are connected by a rigid linker. The overall fold of the catalytic domain of CuGE features a large ten-stranded twisted beta-sheet flanked by alpha-helical elements and a surface-exposed active site. The structure complies with the characteristics of a serine esterase belonging to the alpha/beta-hydrolase superfamily Cerrena unicolor

Synonyms

Synonyms Comment Organism
4-O-methyl-glucuronoyl methylesterase UniProt Cerrena unicolor
CE15 enzyme
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 Cerrena unicolor
CE15-B enzyme
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 Cerrena unicolor
CuGE
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 Cerrena unicolor
glucuronoyl esterase
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 Cerrena unicolor

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
72.2 75.8 inactivation temperatures for all enzyme variants Cerrena unicolor

General Information

General Information Comment Organism
evolution CuGE is an alpha/beta-hydrolase belonging to carbohydrate esterase family 15 (CE15), part of the alpha/beta-hydrolase superfamily. CuGE belongs to the group of fungal CE15-B enzymes with an open and flat substrate-binding site. Conservation of carbohydrate recognition among fungal CE15s. The fungal members of CE15-A display low overall sequence identity compared to the members of CE15-B, furthermore they differ in the configuration of the catalytic triad. The overall sequence identity within the fungal CE15-A group is 40-75%, whereas the sequence identity between the two groups is merely 25-36%. Residues involved in carbohydrate recognition are almost invariant among fungal CE15-B enzymes and most of these residues appear to be conserved in fungal CE15-A members Cerrena unicolor
additional information catalytic domain structure analysis, overview. Wild-type CuGE has a modular architecture with a N-terminal carbohydrate-binding module 1 (CBM1) domain connected to the catalytic domain by a proline-rich linker region. The structure of CuGE is potentially flexible and shows heterogenous N- and O-glycosylation. Active site architecture and ligand-induced oxyanion hole. Structure and substrate binding comparisons of CE15-A and CE15-B enzymes. Detailed structure-function analysis of CE15 enzymes, overview Cerrena unicolor
physiological function the glucuronoyl esterase (GE) from Cerrena unicolor (CuGE) catalyzes cleavage of lignin-carbohydrate ester bonds Cerrena unicolor