TY - JOUR
T1 - Purification and characterization of a ferulic acid esterase (FAE-III) from Aspergillus niger
T2 - Specificity for the phenolic moiety and binding to microcrystalline cellulose
AU - Faulds, C. B.
AU - Williamson, G.
PY - 1994/1/1
Y1 - 1994/1/1
N2 - An inducible ferulic acid esterase (FAE-III) has been isolated, purified and partially characterized from Aspergillus niger after growth on oat spelt xylan. The purification procedure utilized ammonium sulphate precipitation, hydrophobic interaction and anion-exchange chromatography. The purified enzyme appeared almost pure by SDS-PAGE, with an apparent M(r) of 36000. A single band, corresponding to a pl of 3.3 was observed on isoelectric focusing. With methyl ferulate as substrate, the enzyme had a specific activity of 67 IU (mg protein)-1, pH and temperature optima of 5 and 55-60°C, respectively, and a K(m) of 2.08 mM and a V(max) of 175 μmol min-1 (mg protein)-1. The enzyme was also active upon methyl sinapinate, methyl-3,4-dimethoxy cinnamate and methyl p-coumarate, but not benzoic acid methyl esters or methyl caffeate. Similarly, Streptomyces olivochromogenes FAE showed activity against methyl ferulate, methyl sinapinate and methyl p-coumarate, but at a level 420-fold less (on methyl ferulate) than the A. niger esterase. No activity was detected against the benzoate methyl esters. For both enzymes, this shows the necessity for C-3 on the phenol ring to be methoxylated and the aliphatic region of the substrate to be unsaturated. The specific activity of FAE-III on destarched wheat bran was 31 U (mg protein)-1 in the presence of Trichoderma viride xylanase and 3 U (mg protein)-1 in the absence. Apparent pH dependent binding of A. niger FAE-III to microcrystalline cellulose was also demonstrated.
AB - An inducible ferulic acid esterase (FAE-III) has been isolated, purified and partially characterized from Aspergillus niger after growth on oat spelt xylan. The purification procedure utilized ammonium sulphate precipitation, hydrophobic interaction and anion-exchange chromatography. The purified enzyme appeared almost pure by SDS-PAGE, with an apparent M(r) of 36000. A single band, corresponding to a pl of 3.3 was observed on isoelectric focusing. With methyl ferulate as substrate, the enzyme had a specific activity of 67 IU (mg protein)-1, pH and temperature optima of 5 and 55-60°C, respectively, and a K(m) of 2.08 mM and a V(max) of 175 μmol min-1 (mg protein)-1. The enzyme was also active upon methyl sinapinate, methyl-3,4-dimethoxy cinnamate and methyl p-coumarate, but not benzoic acid methyl esters or methyl caffeate. Similarly, Streptomyces olivochromogenes FAE showed activity against methyl ferulate, methyl sinapinate and methyl p-coumarate, but at a level 420-fold less (on methyl ferulate) than the A. niger esterase. No activity was detected against the benzoate methyl esters. For both enzymes, this shows the necessity for C-3 on the phenol ring to be methoxylated and the aliphatic region of the substrate to be unsaturated. The specific activity of FAE-III on destarched wheat bran was 31 U (mg protein)-1 in the presence of Trichoderma viride xylanase and 3 U (mg protein)-1 in the absence. Apparent pH dependent binding of A. niger FAE-III to microcrystalline cellulose was also demonstrated.
KW - Aspergillus niger
KW - Ferulic acid esterase
KW - Streptomyces olivochromogenes
KW - Xylanase
UR - http://www.scopus.com/inward/record.url?scp=0028218853&partnerID=8YFLogxK
U2 - 10.1099/00221287-140-4-779
DO - 10.1099/00221287-140-4-779
M3 - Article
AN - SCOPUS:0028218853
VL - 140
SP - 779
EP - 787
JO - Microbiology-Sgm
JF - Microbiology-Sgm
SN - 1350-0872
IS - 4
ER -