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Literature summary for 3.1.1.117 extracted from
- Structural and biochemical studies of the glucuronoyl esterase OtCE15A illuminate its interaction with lignocellulosic components (2019), J. Biol. Chem., 294, 19978-19987 .
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified OtCE15A S267A mutant in complex with methylgalacturonate and in complex with the glucuronoxylan oligosaccharide XUX, X-ray diffraction structure determination analysis | Opitutus terrae |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D356A | site-directed mutagenesis | Opitutus terrae |
| E290A | site-directed mutagenesis | Opitutus terrae |
| E290A/D356A | site-directed mutagenesis | Opitutus terrae |
| H408A | site-directed mutagenesis of the catalytic His reduces kcat by 1700fold | Opitutus terrae |
| L269Y | site-directed mutagenesis, the mutant shows altered kinetics and substrate specificity compared to wild-type | Opitutus terrae |
| additional information | kinetic characterization of catalytic residue substitutions, structure, ligand interactions, and catalytic mechanism of catalytic OtCE15A variants in complex with GlcA and benzyl glucuronoate, overview | Opitutus terrae |
| S267A | site-directed mutagenesis of the catalytic Ser reduces kcat by 17000fold, the S267A variant of OtCE15A is dramatically crippled | Opitutus terrae |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Opitutus terrae | |
| 0.184 | - |
benzyl-D-glucuronate | mutant H408A, pH and temperature not specified in the publication | Opitutus terrae |  |
| 0.502 | - |
benzyl-D-glucuronate | mutant E290A/D356A, pH and temperature not specified in the publication | Opitutus terrae | |
| 1.69 | - |
methyl-D-glucuronate | mutant L269Y, pH and temperature not specified in the publication | Opitutus terrae | |
| 1.86 | - |
benzyl-D-glucuronate | mutant D356A, pH and temperature not specified in the publication | Opitutus terrae | |
| 2.03 | - |
benzyl-D-glucuronate | mutant E290A, pH and temperature not specified in the publication | Opitutus terrae | |
| 2.77 | - |
methyl-D-glucuronate | wild-type enzyme, pH and temperature not specified in the publication | Opitutus terrae | |
| 2.83 | - |
benzyl-D-glucuronate | mutant S267A, pH and temperature not specified in the publication | Opitutus terrae | |
| 3.57 | - |
benzyl-D-glucuronate | wild-type enzyme, pH and temperature not specified in the publication | Opitutus terrae | |
| 5.31 | - |
methyl-D-galacturonate | wild-type enzyme, pH and temperature not specified in the publication | Opitutus terrae | |
| 11.9 | - |
methyl-D-galacturonate | mutant L269Y, pH and temperature not specified in the publication | Opitutus terrae |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Opitutus terrae | - |
- |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| benzyl-D-glucuronate + H2O | - |
Opitutus terrae | benzyl alcohol + D-glucuronic acid | - |
? | |
| methyl-D-galacturonate + H2O | - |
Opitutus terrae | methanol + galacturonic acid | - |
? | |
| methyl-D-glucuronate + H2O | - |
Opitutus terrae | methanol + glucuronic acid | - |
? | |
| additional information | the enzyme is active with the glucuronoxylan oligosaccharide (XUX). The oligosaccharide is produced from beech xylan and contains the 4-O-methyl-alpha-D-glucuronate moiety. Docking study, structure analysis of enzyme OtCE15A in complex with glucuronoxylooligosaccharide 23-(4-O-methyl-alpha-D-glucuronyl)-xylotriose (commonly referred to as XUX). The structure of the enzyme with the disaccharide xylobiose reveals a surface binding site that possibly indicates a recognition mechanism for long glucuronoxylan chains | Opitutus terrae | ? | - |
- |
|
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| glucuronoyl esterase | - |
Opitutus terrae |
| OtCE15A | - |
Opitutus terrae |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.000983 | - |
benzyl-D-glucuronate | mutant S267A, pH and temperature not specified in the publication | Opitutus terrae | |
| 0.00999 | - |
benzyl-D-glucuronate | mutant H408A, pH and temperature not specified in the publication | Opitutus terrae | |
| 0.196 | - |
benzyl-D-glucuronate | mutant E290A/D356A, pH and temperature not specified in the publication | Opitutus terrae | |
| 2.7 | - |
methyl-D-galacturonate | mutant L269Y, pH and temperature not specified in the publication | Opitutus terrae | |
| 5.21 | - |
benzyl-D-glucuronate | mutant D356A, pH and temperature not specified in the publication | Opitutus terrae | |
| 10.3 | - |
benzyl-D-glucuronate | mutant E290A, pH and temperature not specified in the publication | Opitutus terrae | |
| 16.1 | - |
methyl-D-glucuronate | mutant L269Y, pH and temperature not specified in the publication | Opitutus terrae | |
| 16.6 | - |
benzyl-D-glucuronate | wild-type enzyme, pH and temperature not specified in the publication | Opitutus terrae | |
| 19 | - |
methyl-D-glucuronate | wild-type enzyme, pH and temperature not specified in the publication | Opitutus terrae | |
| 28.8 | - |
methyl-D-galacturonate | wild-type enzyme, pH and temperature not specified in the publication | Opitutus terrae |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | sequence analysis of CE15 has revealed that several members, such as OtCE15A from the soil bacterium Opitutus terrae, have acidic residues at both the canonical and noncanonical positions. OtCE15A is a member of the CE15 family and the alpha/beta-hydrolase superfamily. It features acidic residues at both the canonical and noncanonical positions (Glu290 and Asp356). The observation of CE15 members with acidic residues at both positions may implicate evolutionary transitions between the two positions that could affect interactions with substrates and/or products. All CE15 members contain a catalytic triad comprised of Ser-His-Glu/Asp, as found in other serine-type hydrolases | Opitutus terrae |
| additional information | kinetic characterization of catalytic residue substitutions, structure, ligand interactions, and catalytic mechanism of catalytic OtCE15A variants in complex with GlcA and benzyl glucuronoate, structure analysis of OtCE15A S267A mutant in complex with methylgalacturonate and with the glucuronoxylan oligosaccharide XUX, overview | Opitutus terrae |
| physiological function | enzyme OtCE15A, and likely most of the CE15 family, can utilize esters of glucuronoxylooligosaccharides supporting the proposal that these enzymes work on lignin-carbohydratecomplexes in plant biomass | Opitutus terrae |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00035 | - |
benzyl-D-glucuronate | mutant S267A, pH and temperature not specified in the publication | Opitutus terrae | |
| 0.054 | - |
benzyl-D-glucuronate | mutant H408A, pH and temperature not specified in the publication | Opitutus terrae | |
| 0.23 | - |
methyl-D-galacturonate | mutant L269Y, pH and temperature not specified in the publication | Opitutus terrae | |
| 0.39 | - |
benzyl-D-glucuronate | mutant E290A/D356A, pH and temperature not specified in the publication | Opitutus terrae | |
| 2.8 | - |
benzyl-D-glucuronate | mutant D356A, pH and temperature not specified in the publication | Opitutus terrae | |
| 4.65 | - |
benzyl-D-glucuronate | wild-type enzyme, pH and temperature not specified in the publication | Opitutus terrae | |
| 5.07 | - |
benzyl-D-glucuronate | mutant E290A, pH and temperature not specified in the publication | Opitutus terrae | |
| 5.42 | - |
methyl-D-galacturonate | wild-type enzyme, pH and temperature not specified in the publication | Opitutus terrae | |
| 6.86 | - |
methyl-D-glucuronate | wild-type enzyme, pH and temperature not specified in the publication | Opitutus terrae | |
| 9.53 | - |
methyl-D-glucuronate | mutant L269Y, pH and temperature not specified in the publication | Opitutus terrae |
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