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
| Autor: | Ziyin Yang, Ariaka Murata, Thomas M. Finn, Martha Knight, Susanne Baldermann, Peter Fleischmann, Peter Winterhalter, Andriati N. Mulyadi, Naoharu Watanabe |
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| Rok vydání: | 2011 |
| Předmět: |
Spiral tubing support rotor
Filtration and Separation Centrifugation Fractionation spCCC Analytical Chemistry Dioxygenases Matrix (chemical analysis) chemistry.chemical_compound Ulva Countercurrent chromatography Cleave Acetone Chemical Precipitation Countercurrent Distribution Centrifugal precipitation chromatography CCD Plant Proteins chemistry.chemical_classification Aqueous solution Chromatography Precipitation (chemistry) Protein Seaweed Molecular Weight Enzyme chemistry Chromatography Liquid |
| Zdroj: | Journal of separation science. 34(19) |
| ISSN: | 1615-9314 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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