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Literature summary for 3.1.1.117 extracted from
- Functional expression and characterization of a glucuronoyl esterase from the fungus Neurospora crassa Identification of novel consensus sequences containing the catalytic triad (2016), J. Gen. Appl. Microbiol., 62, 217-224 .
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| Tween 80 | 10 mM, 1.5fold activation | Neurospora crassa |  |
| Tween 80 | activates 50% at 10 mM | Neurospora crassa | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| C-terminally c-myc and hexahistidine-tagged enzyme is heterologously expressed in the methylotrophic yeast Pichia pastoris | Neurospora crassa |
| gene cip2, DNA and amino acid sequence determination and analysis, sequence comparison with Cip2 from Hypocrea jecorina (UniProt ID G0RV93), recombinant expression of C-terminally c-myc- and His6-tagged NcGE from shuttle vector pPICZalphaA in the methylotrophic yeast Pichia pastoris strain KM71H, the enzyme is secreted, enzyme subcloning in Escherichia coli strain DH5alpha | Neurospora crassa |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | 10 mM, 16% loss of activity; 16% inhibition at 10 mM | Neurospora crassa | |
| Acetic anhydride | 10 mM, 7% loss of activity | Neurospora crassa | |
| ascorbic acid | 10 mM, 2.5% loss of activity | Neurospora crassa | |
| Cu2+ | inhibits 37% at 1 mM | Neurospora crassa | |
| CuCl2 | 1 mM, 37% loss of activity | Neurospora crassa | |
| dimethyl sulfoxide | 10 mM, 14% loss of activity; 16% inhibition at 10 mM | Neurospora crassa | |
| dithiothreitol | 10 mM, 29% loss of activity; 29% inhibition at 10 mM | Neurospora crassa | |
| Li+ | inhibits 24% at 1 mM | Neurospora crassa | |
| LiCl | 1 mM, 24% loss of activity | Neurospora crassa | |
| MnCl2 | 1 mM, 6% loss of activity | Neurospora crassa | |
| additional information | the enzyme is not affected by 5 mM EDTA, suggesting that it is not a metallo enzyme | Neurospora crassa | |
| Ni2+ | inhibits 40% at 1 mM | Neurospora crassa | |
| NiSO4 | 1 mM, 40% loss of activity | Neurospora crassa | |
| PMSF | 0.5-1 mM, 59% loss of activity; 60& inhibition at 0.5-1 mM | Neurospora crassa | |
| SDS | 10 mM, complete loss of activity; complete inhibition at 10 mM | Neurospora crassa | |
| Urea | 10 mM, 15% loss of activity | Neurospora crassa | |
| Zn2+ | inhibits 16% at 1 mM | Neurospora crassa | |
| ZnSO4 | 1 mM, 16% loss of activity | Neurospora crassa | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| extracellular | the enzyme is secreted | Neurospora crassa | - |
- |
| extracellular | the enzyme contains an N-terminal signal peptide sequence | Neurospora crassa | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Al3+ | activates 18% at 1 mM | Neurospora crassa | |
| AlCl3 | 1 mM, 1.2fold activation | Neurospora crassa | |
| Ca2+ | activates 27% at 1 mM | Neurospora crassa | |
| CaCl2 | 1 mM, 1.3fold activation | Neurospora crassa | |
| Fe3+ | activates 15% at 1 mM | Neurospora crassa | |
| FeCl3 | 1 mM, 1.1fold activation | Neurospora crassa | |
| Mg2+ | activates 18% at 1 mM, pH 7.0, 30°C, 10 min | Neurospora crassa | |
| MgCl2 | 1 mM, 1.2fold activation | Neurospora crassa | |
| additional information | poor effects on enzyme activity at 1 mM by Mn2+ and Fe2+, and by 5 mM EDTA, 10 mM acetic anhydride, 10 mM ascorbic acid, and 10 mM glycerol | Neurospora crassa |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 41300 | - |
calculated from sequence | Neurospora crassa |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Neurospora crassa | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | ? | - |
- |
|
| additional information | Neurospora crassa CBS 708.71 | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | ? | - |
- |
|
| additional information | Neurospora crassa 74-OR23-1A | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | ? | - |
- |
|
| additional information | Neurospora crassa DSM 1257 | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | ? | - |
- |
|
| additional information | Neurospora crassa ATCC 24698 | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | ? | - |
- |
|
| additional information | Neurospora crassa FGSC 987 | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | ? | - |
- |
|
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Neurospora crassa | Q7S1X0 | - |
- |
| Neurospora crassa 74-OR23-1A | Q7S1X0 | - |
- |
| Neurospora crassa ATCC 24698 | Q7S1X0 | - |
- |
| Neurospora crassa CBS 708.71 | Q7S1X0 | - |
- |
| Neurospora crassa DSM 1257 | Q7S1X0 | - |
- |
| Neurospora crassa FGSC 987 | Q7S1X0 | - |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| no glycoprotein | the genomic sequence encoding NcGE does not contain any intron and the deduced protein lacks the carbohydrate binding module (CBM) | Neurospora crassa |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Neurospora crassa |
| recombinant extracellular c-myc/His6-tagged NcGE from Pichia pastoris, cell culture supernatant by nickel affinity and anion exchange chromatography | Neurospora crassa |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | - |
Neurospora crassa | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | the substrate mimics the ester linkage of 4-O-methyl-D-glucuronic acid in lignin-carbohydrate complexes (LCCs) | Neurospora crassa | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | - |
Neurospora crassa CBS 708.71 | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | the substrate mimics the ester linkage of 4-O-methyl-D-glucuronic acid in lignin-carbohydrate complexes (LCCs) | Neurospora crassa CBS 708.71 | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | - |
Neurospora crassa 74-OR23-1A | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | the substrate mimics the ester linkage of 4-O-methyl-D-glucuronic acid in lignin-carbohydrate complexes (LCCs) | Neurospora crassa 74-OR23-1A | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | - |
Neurospora crassa DSM 1257 | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | the substrate mimics the ester linkage of 4-O-methyl-D-glucuronic acid in lignin-carbohydrate complexes (LCCs) | Neurospora crassa DSM 1257 | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | - |
Neurospora crassa ATCC 24698 | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | the substrate mimics the ester linkage of 4-O-methyl-D-glucuronic acid in lignin-carbohydrate complexes (LCCs) | Neurospora crassa ATCC 24698 | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | - |
Neurospora crassa FGSC 987 | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O | the substrate mimics the ester linkage of 4-O-methyl-D-glucuronic acid in lignin-carbohydrate complexes (LCCs) | Neurospora crassa FGSC 987 | methyl 4-O-methyl-D-glucuronic acid + 3-(4-methoxyphenyl) propyl alcohol | - |
? | |
| additional information | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | Neurospora crassa | ? | - |
- |
|
| additional information | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | Neurospora crassa CBS 708.71 | ? | - |
- |
|
| additional information | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | Neurospora crassa 74-OR23-1A | ? | - |
- |
|
| additional information | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | Neurospora crassa DSM 1257 | ? | - |
- |
|
| additional information | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | Neurospora crassa ATCC 24698 | ? | - |
- |
|
| additional information | the enzyme cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | Neurospora crassa FGSC 987 | ? | - |
- |
|
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 41300, about, sequence calculation, x * 44000, recombinant c-myc- and His6-tagged enzyme, SDS-PAGE | Neurospora crassa |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 4-O-methyl-glucuronoyl methylesterase | UniProt | Neurospora crassa |
| Cip2 | - |
Neurospora crassa |
| NcGE | - |
Neurospora crassa |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | 50 | - |
Neurospora crassa |
| 40 | 50 | recombinant enzyme | Neurospora crassa |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 70 | purified recombinant enzyme NcGE is stable at 30°C, but it completely loses its activity when preincubated at 70°C for 30 min | Neurospora crassa |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
- |
Neurospora crassa |
| 7 | - |
recombinant enzyme | Neurospora crassa |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 4 | 7 | purified recombinant enzyme NcGE is stable in the relatively broad pH range of pH 4.0-7.0 | Neurospora crassa |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Neurospora crassa | sequence calculation | - |
8.8 |
| Neurospora crassa | calculated from sequence | - |
8.8 |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to the C15 family of carbohydrate esterases, the characteristic serine residue in the consensus sequence G-C-S-R-X-G involved in the catalytic mechanism is also conserved in enzyme NcGE. The enzyme harbors another two additional consensus sequences containing the glutamic acid and histidine residues that constitute the catalytic triad of enzyme NcGE | Neurospora crassa |
| additional information | the genomic sequence encoding NcGE does not contain any intron and the deduced protein lacks the carbohydrate binding module (CBM). NcGE has a consensus G-C-S-R-X-G motif conserved in the CE15 family where the serine residue serves as the catalytic nucleophile | Neurospora crassa |
| physiological function | the complete hydrolysis of lignocellulose requires the actions of a variety of enzymes, including those that cleave the linkage between lignin and hemicellulose. The enzyme glucuronoyl esterase cleaves the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | Neurospora crassa |
| physiological function | the complete hydrolysis of lignocellulose requires the actions of a variety of enzymes, including those that cleave the linkage between lignin and hemicellulose. The enzyme glucuronoyl esterase (GE) that constitutes the C15 family of carbohydrate esterases, displays a unique ability to cleave the ester linkage between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid of hemicellulose | Neurospora crassa |
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