title: Application of centrifugal precipitation chromatography and high-speed counter-current chromatography equipped with a spiral tubing support rotor for the isolation and partial characterization of carotenoid cleavage-like enzymes in Enteromorpha compressa (L.) Nees.
abstract: Centrifugal precipitation chromatography and a high-speed counter-current chromatography system equipped with a spiral tubing support rotor (spHSCCC) were successfully applied for the identification and isolation of carotenoid cleavage-like enzymes from Enteromorpha compressa (L.) Nees. This is the first study separating active enzymes from a complex natural matrix by spHSCCC. The target enzymes were identified after fractionation of the proteins in an acetone Tris-buffer gradient by centrifugal precipitation chromatography. Also, an aqueous two-phase solvent system consisting of PEG 1000 and mono- and dibasic potassium phosphate was used for the isolation of the enzymes by spHSCCC. The purified fractions contained two proteins of 65 and 72 kDa, respectively. The enzymes could cleave β-carotene and β-apo-8'-carotenal to produce β-ionone.
authors: Laboratory of Natural Products Chemistry, Graduate School of Science and Technology, Faculty of Agriculture Shizuoka, Shizuoka University, Suruga-ku, Shizuoka, Japan. s_baldermann@yahoo.de
authors: Susanne
authors: S
authors: Baldermann
authors: Andriati N
authors: AN
authors: Mulyadi
authors: Ziyin
authors: Z
authors: Yang
authors: Ariaka
authors: A
authors: Murata
authors: Peter
authors: P
authors: Fleischmann
authors: Peter
authors: P
authors: Winterhalter
authors: Martha
authors: M
authors: Knight
authors: Thomas M
authors: TM
authors: Finn
authors: Naoharu
authors: N
authors: Watanabe
mesh_terms: D002498:Centrifugation / Q000379:methods; D011232:Chemical Precipitation; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D003377:Countercurrent Distribution / Q000295:instrumentation / Q000379:methods*; D049308:Dioxygenases / Q000737:chemistry* / Q000302:isolation & purification*; D008970:Molecular Weight; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D044445:Ulva / Q000737:chemistry / Q000201:enzymology*
chemical_list: D010940:Plant Proteins; D049308:Dioxygenases; C431939:carotenoid cleavage dioxygenase 1