Literature summary for 3.2.1.63 extracted from

Cao, H.; Walton, J.D.; Brumm, P.; Phillips, G.N.
Structure and substrate specificity of a eukaryotic fucosidase from Fusarium graminearum (2014), J. Biol. Chem., 289, 25624-25638.

Cloned(Commentary)

Cloned (Comment)	Organism
sequence comparisons, recombinant enzyme expression of the enzyme including its native signal peptide from pPICZAalpha without the C-terminal Myc/His6 tags in Pichia pastoris, expression of the His6-tagged crystallin domain, residues 501-585, of the enzyme fused to maltose-binding protein at the N-terminus in Escherichia coli	Fusarium graminearum

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified deglycosylated recombinant enzyme in open or closed form or complexed with L-fucose, sitting-drop vapor diffusion method or batch method, several different crystal forms, mixing 0.001 ml of 14-16 mg/ml Endo H-treated fucosidase in 25 mM Tris, pH 7.5, with 0.001 ml of precipitant solution containing 0.1 M Tris, pH 8.0, and either 30% or 40% w/v PEG MME 2000, 20°C, 2-3 days, X-ray diffraction structure determination and analysis at 1.38-156 A resolution, molecular replacement method	Fusarium graminearum

Crystallization (Comment)

Organism

purified deglycosylated recombinant enzyme in open or closed form or complexed with L-fucose, sitting-drop vapor diffusion method or batch method, several different crystal forms, mixing 0.001 ml of 14-16 mg/ml Endo H-treated fucosidase in 25 mM Tris, pH 7.5, with 0.001 ml of precipitant solution containing 0.1 M Tris, pH 8.0, and either 30% or 40% w/v PEG MME 2000, 20°C, 2-3 days, X-ray diffraction structure determination and analysis at 1.38-156 A resolution, molecular replacement method

Fusarium graminearum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	steady-state kinetics of deglycosylated and glycosylated enzyme, overview	Fusarium graminearum

Organism

Organism	UniProt	Comment	Textmining
Fusarium graminearum	J9UN47	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	prediction of three N-glycosylation sites on Asn187, Asn280, and Asn401 (residues numbered for the protein without signal peptide). Deglycosylation of the recombinant enzyme expressed from Pichia pastoris by treatment with Endo H or peptide N-glycosidase F. The N-glycans on Asn-280 of the TIM barrel domain retained a chitobiose core and high mannose type substitutions, GlcNAc-beta1-4GlcNAc-beta1-4(Man-alpha1-6)Man-alpha1-3Man-alpha1-2Man-alpha1-2Man, possibly protected from EndoH by the beta,gamma-crystallin domain. The other two N-glycosylation sites Asn187 and Asn401 are each attached to one GlcNAc residue, indicative of Endo H cleavage. No effect of Endo H deglycosylation on the catalytic efficiency with small fucosylated substrates or on thermal stability, but higher solubility of the glycosylated than the deglycosylated enzyme	Fusarium graminearum

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Pichia pastoris by gel filtration and anion exchange chromatography, recombinant His6-tagged and MBP-fused crystallin domain from Escherichia coli by nickel affinity chromatography	Fusarium graminearum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2'-fucosyllactose + H2O	-	Fusarium graminearum	L-fucose + lactose	-	?
4-nitrophenyl-alpha-L-fucoside + H2O	-	Fusarium graminearum	4-nitrophenol + L-fucose	-	?
additional information	substrate specificity, overview. No activity with 3-fucosyllactose. The enzyme is also active on xyloglucan	Fusarium graminearum	?	-	?

Subunits

Subunits	Comment	Organism
monomer	crystal structure analysis, solution small angle x-ray scattering	Fusarium graminearum

Synonyms

Synonyms	Comment	Organism
FgFCO1	-	Fusarium graminearum

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Fusarium graminearum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5	-	assay at	Fusarium graminearum

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the glycosyl hydrolase family 29, GH29, of retaining fucosidases	Fusarium graminearum
additional information	the enzyme shows active-site conformational flexibility, structure-function analysis, overview	Fusarium graminearum