Premium
Characterisation and purification of a cinnamate esterase from Aspergillus niger industrial pectinase preparation
Author(s) -
Barbe Christine,
Dubourdieu Denis
Publication year - 1998
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/(sici)1097-0010(199812)78:4<471::aid-jsfa141>3.0.co;2-0
Subject(s) - esterase , chemistry , aspergillus niger , hydrolysis , chromatography , biochemistry , cinnamic acid , enzyme
Pectolytic enzymatic preparations used in enology come from Aspergillus niger . They have a desirable principal activity and several secondary activities. Among these secondary activities, three classes of esterases were detected: an esterase specific for ester linkage between cinnamic acid and alcohol (cinnamate esterase); an esterase acting on ester linkage between two phenolic rings (depsidase); and an esterase hydrolysing ester linkage between phenolic acids and straight‐chain alcohols (phenyl esterase). Only the first class of enzyme is able to hydrolyse the cinnamic acid tartrate ester of the white must. The authors tried to purify the cinnamate esterase from the other esterase of an A niger industrial pectolytic preparation. This purification was done in two steps. A gel filtration on S200 HR Sephacryl was first used. This step eluted two fractions: esterases and pectin esterase. The second step was affinity chromatography on a 4B Sepharose EAH grafted with chlorogenic acid. This step eluted two fractions: an unretained fraction containing the phenyl esterase and the depsidase; and a retained fraction with the cinnamate esterase. This enzyme has a molecular weight of 240 kDa and contains two submits of 120 kDa. A new classification for these three esterases is suggested. © 1998 Society of Chemical Industry.