期刊
JOURNAL OF SEPARATION SCIENCE
卷 34, 期 19, 页码 2759-2764出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.201100508
关键词
Centrifugal precipitation chromatography; Counter-current chromatography; Protein; spHSCCC; Spiral tubing support rotor
资金
- Japan Society for the Promotion of Science (JSPS) [07434]
- Grants-in-Aid for Scientific Research [23380063] Funding Source: KAKEN
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 beta-carotene and beta-apo-8'-carotenal to produce beta-ionone.
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