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
✍ Scribed by Susanne Baldermann; Andriati N. Mulyadi; Ziyin Yang; Ariaka Murata; Peter Fleischmann; Peter Winterhalter; Martha Knight; Thomas M. Finn; Naoharu Watanabe
- Publisher
- John Wiley and Sons
- Year
- 2011
- Tongue
- English
- Weight
- 153 KB
- Volume
- 34
- Category
- Article
- ISSN
- 1615-9306
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✦ Synopsis
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.