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Literature summary extracted from

  • Huynh, H.; Ishii, N.; Matsuo, I.; Arioka, M.
    A novel glucuronoyl esterase from Aspergillus fumigatus - the role of conserved Lys residue in the preference for 4-O-methyl glucuronoyl esters (2018), Appl. Microbiol. Biotechnol., 102, 2191-2201 .
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.1.1.117 additional information no significant effect by EDTA, glycerol, acetic anhydride, ascorbic acid, dimethyl sulfoxide, SDS, and urea at 10 mM Aspergillus fumigatus
3.1.1.117 Tween 80 activates 2.3fold at 10 mM Aspergillus fumigatus

Application

EC Number Application Comment Organism
3.1.1.117 industry the enzyme is a potentially industrially applicable enzyme due to its characteristic as a thermophilic enzyme with the favorable temperature of 40-50°C at pH 5 Aspergillus fumigatus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.117 gene Afu6g14390, recombinant expression of Myc-His-tagged enzyme AfGE in Aspergillus oryzae strain RIB40 from vector pUNA-AfGE-Myc-His under the control of a strong promoter of alpha-amylase (amyB), subcloning of the enzyme in Escherichia coli strain DH5alpha and of the vector in Aspergillus oryzae strain niaD300, the recombinant enzyme is secreted into the medium. Recombinant expression of wild-type and mutant enzymes in Pichia pastoris strain KM71H Aspergillus fumigatus
3.1.1.117 heterologously expressed in Aspergillus oryzae RIB40 strain (ATCC42149) under the control of a strong promoter of alpha-amylase (amyB) Aspergillus fumigatus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.117 K209A site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Aspergillus fumigatus
3.1.1.117 K209E site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Aspergillus fumigatus
3.1.1.117 K209Q site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Aspergillus fumigatus
3.1.1.117 K209R site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzymethe mutant shows altered substrate specificity compared to the wild-type enzyme Aspergillus fumigatus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.1.117 2-mercaptoethanol 10 mM Aspergillus fumigatus
3.1.1.117 Al3+ 1 mM, slight inhibition; slight inhibition at 1 mM Aspergillus fumigatus
3.1.1.117 Ca2+ 1 mM, about 40% inhibition; about 40% inhibition at 1 mM Aspergillus fumigatus
3.1.1.117 Cu2+ 1 mM, slight inhibition; slight inhibition at 1 mM Aspergillus fumigatus
3.1.1.117 dithiothreitol 10 mM Aspergillus fumigatus
3.1.1.117 Fe2+ 1 mM, slight inhibition; slight inhibition at 1 mM Aspergillus fumigatus
3.1.1.117 Fe3+ about 40% inhibition at 1 mM Aspergillus fumigatus
3.1.1.117 Li+ 1 mM, slight inhibition; slight inhibition at 1 mM Aspergillus fumigatus
3.1.1.117 Mn2+ 1 mM, slight inhibition; slight inhibition at 1 mM Aspergillus fumigatus
3.1.1.117 additional information AfGE activity is not affected by EDTA, indicating that this enzyme is not a metalloenzyme. No significant effect by glycerol, acetic anhydride, ascorbic acid, dimethyl sulfoxide, SDS, and urea at 10 mM; the enzyme is not affected by EDTA, indicating that this enzyme is not a metalloenzyme Aspergillus fumigatus
3.1.1.117 Ni2+ 1 mM, slight inhibition; about 40% inhibition at 1 mM Aspergillus fumigatus
3.1.1.117 PMSF 10 mM Aspergillus fumigatus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.117 15.8
-
 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate pH 5.0, 30°C Aspergillus fumigatus
3.1.1.117 16.4
-
 benzyl D-glucuronate pH 5.0, 30°C Aspergillus fumigatus
3.1.1.117 23.7
-
 benzyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate pH 5.0, 30°C Aspergillus fumigatus
3.1.1.117 38
-
 benzyl methyl alpha-D-glucopyranosiduronate pH 5.0, 30°C Aspergillus fumigatus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.1.1.117 extracellular
-
 Aspergillus fumigatus
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.1.117 Mg2+ 1 mM, increases activity Aspergillus fumigatus
3.1.1.117 Mg2+ activates at 1 mM Aspergillus fumigatus
3.1.1.117 additional information the enzyme is not affected by Zn2+ at 1 mM. AfGE activity is not affected by EDTA, indicating that this enzyme is not a metalloenzyme Aspergillus fumigatus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.1.117 44000
-
 calculated, enzyme with a Myc-His tag added to the C-terminal end of AfGE by the fusion PCR Aspergillus fumigatus

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.117 Aspergillus fumigatus Q4WL89
-
-
3.1.1.117 Aspergillus fumigatus Q4WL89 i.e. Aspergillus fumigatus
-
3.1.1.117 Aspergillus fumigatus ATCC MYA-4609 Q4WL89
-
-
3.1.1.117 Aspergillus fumigatus ATCC MYA-4609 Q4WL89 i.e. Aspergillus fumigatus
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.1.1.117 glycoprotein the enzyme has two potential N-glycosylation sites Aspergillus fumigatus

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.117
-
 Aspergillus fumigatus
3.1.1.117 recombinant Myc-His-tagged enzyme AfGE from Aspergillus oryzae strain RIB40 by nickel affinity chromatography, and anion exchange chromatography Aspergillus fumigatus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.117 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O
-
 Aspergillus fumigatus 3-(4-methoxyphenyl)propan-1-ol + methyl 4-O-methyl-alpha-D-glucopyranosiduronic acid
-
 ?
3.1.1.117 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O
-
 Aspergillus fumigatus ATCC MYA-4609 3-(4-methoxyphenyl)propan-1-ol + methyl 4-O-methyl-alpha-D-glucopyranosiduronic acid
-
 ?
3.1.1.117 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O the substrate mimics the ester linkage between lignin and hemicellulose Aspergillus fumigatus methanol + 3-(4-methoxyphenyl) propyl 4-O-methyl-alpha-D-glucopyranosiduronate
-
 ?
3.1.1.117 benzyl D-glucuronate + H2O
-
 Aspergillus fumigatus benzyl alcohol + D-glucuronic acid
-
 ?
3.1.1.117 benzyl D-glucuronate + H2O
-
 Aspergillus fumigatus ATCC MYA-4609 benzyl alcohol + D-glucuronic acid
-
 ?
3.1.1.117 benzyl D-glucuronate + H2O
-
 Aspergillus fumigatus benzyl alcohol + glucuronic acid
-
 ?
3.1.1.117 benzyl D-glucuronate + H2O
-
 Aspergillus fumigatus ATCC MYA-4609 benzyl alcohol + glucuronic acid
-
 ?
3.1.1.117 benzyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O
-
 Aspergillus fumigatus benzyl alcohol + methyl 4-O-methyl-alpha-D-glucopyranosiduronic acid
-
 ?
3.1.1.117 benzyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate + H2O
-
 Aspergillus fumigatus ATCC MYA-4609 benzyl alcohol + methyl 4-O-methyl-alpha-D-glucopyranosiduronic acid
-
 ?
3.1.1.117 benzyl methyl alpha-D-glucopyranosiduronate + H2O
-
 Aspergillus fumigatus benzyl alcohol + methyl alpha-D-glucopyranosiduronic acid
-
 ?
3.1.1.117 additional information Lys209 plays an important role in the preference for the substrates containing 4-O-methyl group in the glucopyranose ring. The purified recombinant enzyme displays the ability to degrade the synthetic substrates mimicking the ester linkage between hemicellulose and lignin. Self-degradation of the substrates under high alkaline condition Aspergillus fumigatus ?
-
-
3.1.1.117 additional information Lys209 plays an important role in the preference for the substrates containing 4-O-methyl group in the glucopyranose ring. The purified recombinant enzyme displays the ability to degrade the synthetic substrates mimicking the ester linkage between hemicellulose and lignin. Self-degradation of the substrates under high alkaline condition Aspergillus fumigatus ATCC MYA-4609 ?
-
-

Subunits

EC Number Subunits Comment Organism
3.1.1.117 ? x * 44000, recombinant deglycosylated Myc-His-tagged enzyme AfGE, SDS-PAGE Aspergillus fumigatus

Synonyms

EC Number Synonyms Comment Organism
3.1.1.117 AfGE
-
 Aspergillus fumigatus
3.1.1.117 Afu6g14390
-
 Aspergillus fumigatus
3.1.1.117 Cip2
-
 Aspergillus fumigatus
3.1.1.117 glucuronoyl esterase
-
 Aspergillus fumigatus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.117 40
-
-
 Aspergillus fumigatus
3.1.1.117 40
-
 recombinant enzyme Aspergillus fumigatus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.1.1.117 30 70 30°C: about 80% of maximal activity, 70°C: about 60% of maximal activity Aspergillus fumigatus
3.1.1.117 30 65 over 50% of maximal activity within this range, profile overview Aspergillus fumigatus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.117 30 50 30 min, pH 7.0, stable Aspergillus fumigatus
3.1.1.117 30 50 purified recombinant enzyme, pH 7.0, 30 min, completely stable at Aspergillus fumigatus
3.1.1.117 60
-
 30 min, pH 7.0, about 30% loss of activity Aspergillus fumigatus
3.1.1.117 70
-
 30 min, pH 7.0, about 70% loss of activity Aspergillus fumigatus
3.1.1.117 70
-
 purified recombinant enzyme, pH 7.0, 30 min, loss of 60% activity Aspergillus fumigatus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.1.117 23.8
-
 benzyl D-glucuronate pH 5.0, 30°C Aspergillus fumigatus
3.1.1.117 42.7
-
 benzyl methyl alpha-D-glucopyranosiduronate pH 5.0, 30°C Aspergillus fumigatus
3.1.1.117 66.4
-
 benzyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate pH 5.0, 30°C Aspergillus fumigatus
3.1.1.117 67.1
-
 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate pH 5.0, 30°C Aspergillus fumigatus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.117 5
-
-
 Aspergillus fumigatus
3.1.1.117 5
-
 recombinant enzyme Aspergillus fumigatus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.1.117 4 8 over 60% of maximal activity within this range, profile overview Aspergillus fumigatus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.1.1.117 3 9 purified recombinant enzyme, 30°C, 30 min, over 40% of maximal activity remaining within this range, 80% at pH 5.0 Aspergillus fumigatus
3.1.1.117 4
-
 30°C, 30 min, activity is significantly lost after pre-incubation in pHs lower than 4 Aspergillus fumigatus
3.1.1.117 5 7 30°C, 30 min, high stability Aspergillus fumigatus
3.1.1.117 8
-
 30°C, 30 min, activity is significantly lost after pre-incubation in pHs higher than 8 Aspergillus fumigatus

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.1.1.117 Aspergillus fumigatus sequence calculation
-
 6.1

General Information

EC Number General Information Comment Organism
3.1.1.117 evolution glucuronoyl esterases (GEs) belong to the family 15 of carbohydrate esterases (CE15). Sequence comparison with enzyme Cip2 (UniProt ID G0RV93) from Hypocrea jecorina strain QM6a Aspergillus fumigatus
3.1.1.117 additional information role of conserved Lys209 residue in the preference for 4-O-methyl glucuronoyl esters, AfGE docking study and structure homology molecular modeling using the crystal structure of StGE2 S213A mutant docked with methyl 4-O-methyl-beta-D-glucopyranuronate (PDB ID 4G4J) as the template, overview Aspergillus fumigatus
3.1.1.117 physiological function cellulose in plant cell walls is mainly covered by hemicellulose and lignin, and thus efficient removal of these components is thought to be a key step in the optimal utilization of lignocellulose. The carbohydrate esterase (CE) 15 family of glucuronoyl esterases (GEs), which cleave the linkages between the free carboxyl group of D-glucuronic acid in hemicellulose and the benzyl groups in lignin residues, can contribute to this process Aspergillus fumigatus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.1.117 1.12
-
 benzyl methyl alpha-D-glucopyranosiduronate pH 5.0, 30°C Aspergillus fumigatus
3.1.1.117 1.45
-
 benzyl D-glucuronate pH 5.0, 30°C Aspergillus fumigatus
3.1.1.117 2.8
-
 benzyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate pH 5.0, 30°C Aspergillus fumigatus
3.1.1.117 4.24
-
 3-(4-methoxyphenyl) propyl methyl 4-O-methyl-alpha-D-glucopyranosiduronate pH 5.0, 30°C Aspergillus fumigatus