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Protein extraction using sugar ester reverse micelles
Author(s) -
Naoe Kazumitsu,
Nishino Makiko,
Ohsa Tomomi,
Kawagoe Mikio,
Imai Masanao
Publication year - 1999
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/(sici)1097-4660(199903)74:3<221::aid-jctb47>3.0.co;2-o
Subject(s) - chemistry , micelle , extraction (chemistry) , critical micelle concentration , hexane , chromatography , isopropyl alcohol , pulmonary surfactant , lecithin , aqueous solution , sugar , organic chemistry , biochemistry
In protein extraction, using the nonionic surfactant sugar ester DK‐F‐110, the critical micelle concentration (CMC) of a DK‐F‐110/isopropyl alcohol(IPA)/hexane system was found to be at a DK‐F‐110 concentration of 0.5 g dm −3 , indicating that the sugar ester reverse micelles could be formed in hexane. At concentrations higher than the CMC, cytochrome c was extracted into the DK‐F110 reverse micelles as judged from UV spectra of the DK‐F‐110/IPA/hexane solution after it was contacted with the aqueous protein solution. In extraction of cytochrome c using this system, forwardextraction was found to be pH‐dependent, with high extraction percentage being obtained at pH 8. The forward extraction percentage was reduced by an increase in the buffer concentration, and at a buffer concentration of 0.5 mol dm −3 was ca 25% as high as that of sodium bis(2‐ethylhexyl) sulfosuccinate(AOT) systems. An optimal DK‐F‐110 concentration was found to give the maximum forward extraction percentage. Addition of alcohol, especially IPA, to the micellar organic phase enabled the highly backward extraction of cytochrome c to be achieved without the formation of insoluble aggregates. The esterification reaction rate by Rhizopus delemar lipase in the DK‐F‐110 reverse micellar system had a higher maximum value than that of AOT and lecithin systems. © 1999 Society of Chemical Industry