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Literature summary extracted from

  • Arnling Baath, J.; Mazurkewich, S.; Poulsen, J.N.; Olsson, L.; Lo Leggio, L.; Larsbrink, J.
    Structure-function analyses reveal that a glucuronoyl esterase from Teredinibacter turnerae interacts with carbohydrates and aromatic compounds (2019), J. Biol. Chem., 294, 6635-6644 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
3.1.1.117 degradation Glucuronoyl esterases (GEs) catalyze the cleavage of ester linkages found between lignin and glucuronic acid moieties on glucuronoxylan in plant biomass. As such, GEs represent promising biochemical tools in industrial processing of these recalcitrant resources Teredinibacter turnerae

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.117 recombinant expression of His-tagged wild-type and mutant enzymes Teredinibacter turnerae

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.1.117 purified recombinant enzyme TtCE15A, sitting drop vapour diffusion method, mixing of protein:reservoir volume ratios of 3:1 or 1:1 using 35 mg/ml protein in 20 mM Tris, pH 8.0, and reservoir solution containing 0.09 M halogens, 0.1 M imidazole and MES buffer, and 50% v/v precipitant mixture 2 containing 40% v/v ethylene glycol and 20% w/v PEG 8000, X-ray diffraction structure determination and analysis at 2.5 A, molecular replacement and model building Teredinibacter turnerae

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.117 E374A site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme Teredinibacter turnerae
3.1.1.117 F174A site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme Teredinibacter turnerae
3.1.1.117 F174D site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme Teredinibacter turnerae
3.1.1.117 S281A site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme Teredinibacter turnerae
3.1.1.117 S304E site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme Teredinibacter turnerae
3.1.1.117 S304E/E374A site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme Teredinibacter turnerae
3.1.1.117 W376A site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme Teredinibacter turnerae
3.1.1.117 W376D site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme Teredinibacter turnerae

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.1.117 4-coumaric acid
-
 Teredinibacter turnerae
3.1.1.117 ferulic acid
-
 Teredinibacter turnerae
3.1.1.117 reduced 23-(4-O-methyl-D-glucuronyl)-alpha-D-xylotetraitol XUXXR, inhibits the wild-type, mutant F174D, and mutant F174A, but not mutant W376A Teredinibacter turnerae
3.1.1.117 Sinapic acid
-
 Teredinibacter turnerae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.117 3.47
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, recombinant His-tagged enzyme Teredinibacter turnerae
3.1.1.117 4
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant S281A Teredinibacter turnerae
3.1.1.117 12.7
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant S304E/E374A Teredinibacter turnerae
3.1.1.117 15.4
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant W376D Teredinibacter turnerae
3.1.1.117 21.8
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant E374A Teredinibacter turnerae
3.1.1.117 22.7
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant W376A Teredinibacter turnerae
3.1.1.117 25.8
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant F174A Teredinibacter turnerae
3.1.1.117 29.4
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant S304E Teredinibacter turnerae
3.1.1.117 49
-
 allyl-D-glucuronate pH 8.5, temperature not specified in the publication, recombinant His-tagged enzyme Teredinibacter turnerae
3.1.1.117 53.1
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant F174D Teredinibacter turnerae

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.117 Teredinibacter turnerae C5BN23 Teredinibacter turnerae encodes three CE15 enzymes: TtCE15A, -B, and -C, locus tags are TERTU_0517, TERTU_3447, and TERTU_3514, respectively
-
3.1.1.117 Teredinibacter turnerae ATCC 39867 C5BN23 Teredinibacter turnerae encodes three CE15 enzymes: TtCE15A, -B, and -C, locus tags are TERTU_0517, TERTU_3447, and TERTU_3514, respectively
-
3.1.1.117 Teredinibacter turnerae T7901 C5BN23 Teredinibacter turnerae encodes three CE15 enzymes: TtCE15A, -B, and -C, locus tags are TERTU_0517, TERTU_3447, and TERTU_3514, respectively
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.117 recombinant His-tagged wild-type and mutant enzymes by nickel affinity chromatography Teredinibacter turnerae

Reaction

EC Number Reaction Comment Organism Reaction ID
3.1.1.117 a 4-O-methyl-D-glucopyranuronate ester + H2O = 4-O-methyl-D-glucuronic acid + an alcohol The serine and histidine residues of the catalytic triad found in all solved GE structures (fungal and bacterial) are conserved in TtCE15A: Ser281 and His427, only the proposed canonical glutamate of the catalytic triad is absent (similar to MZ0003), and this position is occupied by a serine residue, Ser304. TtCE15A is fine-tuned to utilize Glu374 as the acidic residue in the catalytic mechanism, supporting the enzyme's high turnover rate, and that the utilization of the residue in this position is distinct from CE15 members exhibiting the canonical acidic residue Teredinibacter turnerae
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.117 allyl-D-glucuronate + H2O low activity Teredinibacter turnerae prop-2-en-1-ol + glucuronic acid
-
 ?
3.1.1.117 allyl-D-glucuronate + H2O low activity Teredinibacter turnerae ATCC 39867 prop-2-en-1-ol + glucuronic acid
-
 ?
3.1.1.117 allyl-D-glucuronate + H2O low activity Teredinibacter turnerae T7901 prop-2-en-1-ol + glucuronic acid
-
 ?
3.1.1.117 benzyl-D-glucuronate + H2O
-
 Teredinibacter turnerae benzyl alcohol + D-glucuronic acid
-
 ?
3.1.1.117 benzyl-D-glucuronate + H2O
-
 Teredinibacter turnerae ATCC 39867 benzyl alcohol + D-glucuronic acid
-
 ?
3.1.1.117 benzyl-D-glucuronate + H2O
-
 Teredinibacter turnerae T7901 benzyl alcohol + D-glucuronic acid
-
 ?
3.1.1.117 methyl-D-galacturonate + H2O very low activity Teredinibacter turnerae methanol + galacturonic acid
-
 ?
3.1.1.117 methyl-D-galacturonate + H2O very low activity Teredinibacter turnerae ATCC 39867 methanol + galacturonic acid
-
 ?
3.1.1.117 methyl-D-galacturonate + H2O very low activity Teredinibacter turnerae T7901 methanol + galacturonic acid
-
 ?
3.1.1.117 methyl-D-glucuronate + H2O very low activity Teredinibacter turnerae methanol + glucuronic acid
-
 ?
3.1.1.117 methyl-D-glucuronate + H2O very low activity Teredinibacter turnerae ATCC 39867 methanol + glucuronic acid
-
 ?
3.1.1.117 methyl-D-glucuronate + H2O very low activity Teredinibacter turnerae T7901 methanol + glucuronic acid
-
 ?
3.1.1.117 additional information glucuronoyl esterase from Teredinibacter turnerae interacts with carbohydrates and aromatic compounds, enzyme-substrate interaction analysis, two aromatic residues, Phe174 and Trp376, conserved in bacterial GEs, interact with aromatic and carbohydrate structures of these substrates in the enzyme active site, respectively. The substrate affinity decreases drastically for glucuronoate esters with smaller alcohol portions (allyl and methyl) as compared to benzyl-D-glucuronate. Poor activity with 4-nitrophenylacetate Teredinibacter turnerae ?
-
-
3.1.1.117 additional information glucuronoyl esterase from Teredinibacter turnerae interacts with carbohydrates and aromatic compounds, enzyme-substrate interaction analysis, two aromatic residues, Phe174 and Trp376, conserved in bacterial GEs, interact with aromatic and carbohydrate structures of these substrates in the enzyme active site, respectively. The substrate affinity decreases drastically for glucuronoate esters with smaller alcohol portions (allyl and methyl) as compared to benzyl-D-glucuronate. Poor activity with 4-nitrophenylacetate Teredinibacter turnerae ATCC 39867 ?
-
-
3.1.1.117 additional information glucuronoyl esterase from Teredinibacter turnerae interacts with carbohydrates and aromatic compounds, enzyme-substrate interaction analysis, two aromatic residues, Phe174 and Trp376, conserved in bacterial GEs, interact with aromatic and carbohydrate structures of these substrates in the enzyme active site, respectively. The substrate affinity decreases drastically for glucuronoate esters with smaller alcohol portions (allyl and methyl) as compared to benzyl-D-glucuronate. Poor activity with 4-nitrophenylacetate Teredinibacter turnerae T7901 ?
-
-

Synonyms

EC Number Synonyms Comment Organism
3.1.1.117 carbohydrate esterase family 15 domain protein UniProt Teredinibacter turnerae
3.1.1.117 glucuronoyl esterase
-
 Teredinibacter turnerae
3.1.1.117 TERTU_0517 locus name Teredinibacter turnerae
3.1.1.117 TtCE15A
-
 Teredinibacter turnerae

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.117 52 54 differential scanning fluorimetry confirms similar melting temperatures (Tm) for the Phe174 variant as 52-54°C, and for the wild-type enzyme as 52°C Teredinibacter turnerae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.1.117 0.012
-
 methyl-D-galacturonate pH 8.5, temperature not specified in the publication, recombinant His-tagged enzyme Teredinibacter turnerae
3.1.1.117 0.36
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant S281A Teredinibacter turnerae
3.1.1.117 1.92
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant S304E Teredinibacter turnerae
3.1.1.117 2.83
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant S304E/E374A Teredinibacter turnerae
3.1.1.117 4.12
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant E374A Teredinibacter turnerae
3.1.1.117 12.8
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant W376D Teredinibacter turnerae
3.1.1.117 18.3
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant W376A Teredinibacter turnerae
3.1.1.117 43.1
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant F174D Teredinibacter turnerae
3.1.1.117 132
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, recombinant His-tagged enzyme Teredinibacter turnerae
3.1.1.117 136
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant F174A Teredinibacter turnerae
3.1.1.117 200
-
 allyl-D-glucuronate pH 8.5, temperature not specified in the publication, recombinant His-tagged enzyme Teredinibacter turnerae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.117 8.5
-
 TtCE15A Teredinibacter turnerae

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.1.1.117 0.084
-
 pH 8.5, temperature not specified in the publication, recombinant mutant F174A Teredinibacter turnerae ferulic acid
3.1.1.117 0.19
-
 pH 8.5, temperature not specified in the publication, recombinant mutant F174A Teredinibacter turnerae Sinapic acid
3.1.1.117 0.4
-
 pH 8.5, temperature not specified in the publication, recombinant mutant F174D Teredinibacter turnerae ferulic acid
3.1.1.117 0.55
-
 pH 8.5, temperature not specified in the publication, recombinant wild-type enzyme Teredinibacter turnerae ferulic acid
3.1.1.117 0.56
-
 pH 8.5, temperature not specified in the publication, recombinant wild-type enzyme Teredinibacter turnerae Sinapic acid
3.1.1.117 0.77
-
 pH 8.5, temperature not specified in the publication, recombinant mutant F174D Teredinibacter turnerae Sinapic acid
3.1.1.117 0.83
-
 pH 8.5, temperature not specified in the publication, recombinant wild-type enzyme Teredinibacter turnerae 4-coumaric acid
3.1.1.117 4.84
-
 pH 8.5, temperature not specified in the publication, recombinant mutant F174A Teredinibacter turnerae 4-coumaric acid
3.1.1.117 5.51
-
 pH 8.5, temperature not specified in the publication, recombinant wild-type enzyme Teredinibacter turnerae reduced 23-(4-O-methyl-D-glucuronyl)-alpha-D-xylotetraitol
3.1.1.117 26
-
 pH 8.5, temperature not specified in the publication, recombinant mutant F174D Teredinibacter turnerae 4-coumaric acid

General Information

EC Number General Information Comment Organism
3.1.1.117 evolution the enzyme belongs to the carbohydrate esterase family 15 Teredinibacter turnerae
3.1.1.117 additional information enzyme active site structure and structure comparisons, overview. TtCE15A has an alpha/beta-hydrolase fold similar to other CE15 structures, consisting of a three-layer sandwich with a solvent-exposed cleft comprising the active site with its catalytic triad. Three TtCE15A molecules are found in the asymmetric unit, with a Calpha root mean square deviation below 0.2A, indicating a high degree of similarity between the protein chains. The serine and histidine residues of the catalytic triad found in all solved GE structures (fungal and bacterial) are conserved in TtCE15A: Ser281 and His427, only the proposed canonical glutamate of the catalytic triad is absent (similar to MZ0003), and this position is occupied by a serine residue, Ser304. TtCE15A is fine-tuned to utilize Glu374 as the acidic residue in the catalytic mechanism, supporting the enzyme's high turnover rate, and that the utilization of the residue in this position is distinct from CE15 members exhibiting the canonical acidic residue Teredinibacter turnerae

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 0.065
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant S304E Teredinibacter turnerae
3.1.1.117 0.09
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant S281A Teredinibacter turnerae
3.1.1.117 0.189
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant E374A Teredinibacter turnerae
3.1.1.117 0.223
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant S304E/E374A Teredinibacter turnerae
3.1.1.117 0.806
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant W376A Teredinibacter turnerae
3.1.1.117 0.811
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant F174D Teredinibacter turnerae
3.1.1.117 0.831
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant W376D Teredinibacter turnerae
3.1.1.117 4.08
-
 allyl-D-glucuronate pH 8.5, temperature not specified in the publication, recombinant His-tagged enzyme Teredinibacter turnerae
3.1.1.117 5.27
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, mutant F174A Teredinibacter turnerae
3.1.1.117 38
-
 benzyl-D-glucuronate pH 8.5, temperature not specified in the publication, recombinant His-tagged enzyme Teredinibacter turnerae