Literature summary for 3.1.1.73 extracted from

Faulds, C.B.; Molina, R.; Gonzalez, R.; Husband, F.; Juge, N.; Sanz-Aparicio, J.; Hermoso, J.A.
Probing the determinants of substrate specificity of a feruloyl esterase, AnFaeA, from Aspergillus niger (2005), FEBS J., 272, 4362-4371.

Cloned(Commentary)

Cloned (Comment)	Organism
Y80 and W260 mutants expressed in Pichia pastoris	Aspergillus niger

Crystallization (Commentary)

Crystallization (Comment)	Organism
S133A mutant in complex with O-(5-O-[(E)-feruloyl]-alpha-L-arabinofuranosyl)-(1,3)-O-beta-D-xylopyranosyl-(1,4)-D-xylopyranose, hanging drop vapor diffusion method	Aspergillus niger

Protein Variants

Protein Variants	Comment	Organism
S133A	inactive	Aspergillus niger
W260S	significantly reduced turnover, increase in alpha-helix content, broadening of substrate specificity allowing the hydrolysis of methyl caffeate	Aspergillus niger
W260V	significantly reduced turnover, increase in alpha-helix content, broadening of substrate specificity allowing the hydrolysis of methyl caffeate	Aspergillus niger
Y80S	significantly reduced turnover, increase in alpha-helix content, broadening of substrate specificity allowing the hydrolysis of methyl caffeate	Aspergillus niger
Y80V	significantly reduced turnover, increase in alpha-helix content, broadening of substrate specificity allowing the hydrolysis of methyl caffeate	Aspergillus niger

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.24	-	methyl sinapinate	pH 6.0, 37°C, native enzyme	Aspergillus niger
0.35	-	methyl sinapinate	pH 6.0, 37°C, W260S mutant enzyme	Aspergillus niger
0.76	-	methyl sinapinate	pH 6.0, 37°C, W260V mutant enzyme	Aspergillus niger
0.78	-	methyl ferulate	pH 6.0, 37°C, native enzyme	Aspergillus niger
0.84	-	methyl sinapinate	pH 6.0, 37°C, Y80S mutant enzyme	Aspergillus niger
0.88	-	methyl ferulate	pH 6.0, 37°C, W260S mutant enzyme	Aspergillus niger
1.01	-	methyl ferulate	pH 6.0, 37°C, W260V mutant enzyme	Aspergillus niger
1.17	-	methyl ferulate	pH 6.0, 37°C, Y80V mutant enzyme	Aspergillus niger
1.22	-	methyl ferulate	pH 6.0, 37°C, Y80S mutant enzyme	Aspergillus niger
1.23	-	methyl sinapinate	pH 6.0, 37°C, Y80V mutant enzyme	Aspergillus niger
2.07	-	methyl p-coumarate	pH 6.0, 37°C, Y80S mutant enzyme	Aspergillus niger
2.88	-	methyl p-coumarate	pH 6.0, 37°C, Y80V mutant enzyme	Aspergillus niger
3.02	-	methyl caffeate	pH 6.0, 37°C, Y80V mutant enzyme	Aspergillus niger
3.26	-	methyl p-coumarate	pH 6.0, 37°C, W260S mutant enzyme	Aspergillus niger
3.84	-	methyl caffeate	pH 6.0, 37°C, Y80S mutant enzyme	Aspergillus niger
3.99	-	methyl p-coumarate	pH 6.0, 37°C, W260V mutant enzyme	Aspergillus niger
4.1	-	methyl caffeate	pH 6.0, 37°C, W260S mutant enzyme	Aspergillus niger
4.26	-	methyl p-coumarate	pH 6.0, 37°C, native enzyme	Aspergillus niger
4.72	-	methyl caffeate	pH 6.0, 37°C, W260V mutant enzyme	Aspergillus niger

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
extracellular	-	Aspergillus niger	-	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
feruloyl polysaccharide + H2O	Aspergillus niger	involved in the degradation of plant cell wall material, breaks ferulic acid cross-links between cell wall components	ferulic acid + ?	-	?

Organism

Organism	UniProt	Comment	Textmining
Aspergillus niger	-	type A enzyme	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
feruloyl polysaccharide + H2O	involved in the degradation of plant cell wall material, breaks ferulic acid cross-links between cell wall components	Aspergillus niger	ferulic acid + ?	-	?
methyl caffeate + H2O	no activity of native enzyme, mutant enzymes are capable in hydrolyzing methyl caffeate	Aspergillus niger	caffeic acid + methanol	-	?
methyl ferulate + H2O	-	Aspergillus niger	ferulic acid + methanol	-	?
methyl p-coumarate + H2O	-	Aspergillus niger	p-coumaric acid + methanol	-	?
methyl sinapinate + H2O	-	Aspergillus niger	sinapic acid + methanol	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.01	-	methyl caffeate	pH 6.0, 37°C, Y80S mutant enzyme	Aspergillus niger
0.02	-	methyl caffeate	pH 6.0, 37°C, Y80V mutant enzyme	Aspergillus niger
0.031	0.51	methyl sinapinate	pH 6.0, 37°C, Y80V mutant enzyme	Aspergillus niger
0.1	-	methyl p-coumarate	pH 6.0, 37°C, Y80S mutant enzyme	Aspergillus niger
0.26	-	methyl caffeate	pH 6.0, 37°C, W260V mutant enzyme	Aspergillus niger
0.26	-	methyl p-coumarate	pH 6.0, 37°C, Y80V mutant enzyme	Aspergillus niger
0.29	-	methyl p-coumarate	pH 6.0, 37°C, W260V mutant enzyme	Aspergillus niger
0.32	-	methyl caffeate	pH 6.0, 37°C, W260S mutant enzyme	Aspergillus niger
0.49	-	methyl p-coumarate	pH 6.0, 37°C, W260S mutant enzyme	Aspergillus niger
0.73	-	methyl p-coumarate	pH 6.0, 37°C, native enzyme	Aspergillus niger
0.99	-	methyl ferulate	pH 6.0, 37°C, W260V mutant enzyme	Aspergillus niger
1.56	-	methyl ferulate	pH 6.0, 37°C, Y80S mutant enzyme	Aspergillus niger
1.6	-	methyl ferulate	pH 6.0, 37°C, native enzyme	Aspergillus niger
2	3.7	methyl sinapinate	pH 6.0, 37°C, native enzyme	Aspergillus niger
2.56	-	methyl ferulate	pH 6.0, 37°C, Y80V mutant enzyme	Aspergillus niger
3.48	-	methyl sinapinate	pH 6.0, 37°C, Y80S mutant enzyme	Aspergillus niger
3.5	-	methyl ferulate	pH 6.0, 37°C, W260S mutant enzyme	Aspergillus niger
7.85	-	methyl sinapinate	pH 6.0, 37°C, Y80V mutant enzyme	Aspergillus niger
18.33	-	methyl ferulate	pH 6.0, 37°C, W260V mutant enzyme	Aspergillus niger
20.06	-	methyl ferulate	pH 6.0, 37°C, W260S mutant enzyme	Aspergillus niger
27.76	-	methyl sinapinate	pH 6.0, 37°C, W260S mutant enzyme	Aspergillus niger
28.28	-	methyl sinapinate	pH 6.0, 37°C, W260V mutant enzyme	Aspergillus niger
70.74	-	methyl ferulate	pH 6.0, 37°C, native enzyme	Aspergillus niger
84.95	-	methyl sinapinate	pH 6.0, 37°C, native enzyme	Aspergillus niger