title: Purification and characterization of cis-aconitic acid decarboxylase from Aspergillus terreus TN484-M1.
abstract: cis-Aconitic acid decarboxylase (CAD) was assumed to be a key enzyme in the production of itaconic acid by comparing the activity of CAD from Aspergillus terreus TN484-M1 with that of CAD from the low-itaconate yielding strain Aspergillus terreus CM85J. The constitutive CAD was purified to homogeneity from A. terreus TN484-M1 by ammonium sulfate fractionation, and column chromatography on DEAE-toyopearl, Butyl-toyopearl, and Sephacryl S200HR, and then characterized. A molecular mass of 55 kDa for the native enzyme was determined by SDS-PAGE. The enzymic activity was optimal at a pH of 6.2 and temperature of 45 degrees C. The K(m) value for cis-aconitic acid was determined as 2.45 mM (pH 6.2, 37 degrees C). The enzyme was completely inactivated by Hg+, Cu2+, Zn2+, p-chloromercuribenzoate, and 5,5'-dithio-bis(2-nitrobenzoate).
authors.affiliation: Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka-shi 422-8529, Japan.
authors.forename: Lies
authors.initials: L
authors.lastname: Dwiarti
authors.forename: Ken
authors.initials: K
authors.lastname: Yamane
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Yamatani
authors.forename: Prihardi
authors.initials: P
authors.lastname: Kahar
authors.forename: Mitsuyasu
authors.initials: M
authors.lastname: Okabe