Add time:09/04/2019         Source:sciencedirect.com

In protein extraction using non-ionic Span 60 reverse micelles, the critical micelle concentration (CMC) of a Span 60/IPA/hexane system was at Span 60 concentration of 3.5 g l−1. At concentrations higher than the CMC, cytochrome c was extracted into the Span 60 reverse micelles as judged from UV spectra of the Span 60/IPA/hexane solution after it contacted with the protein aqueous solution. In extractions of cytochrome c and lysozyme using this system, forward extraction was found to be pH-dependent, with high extraction percentages being obtained at neutral pHs. The forward extraction percentage was drastically reduced by only a small increase in the buffer concentration (even up to 0.1 M). In cytochrome c extraction using various means of phase contacting, high extraction percentages were obtained with rotary and reciprocal shaking, but in well-mixed extraction by magnetic stirring, extraction percentages were much lower. Addition of alcohol, especially isopropyl alcohol, to the micellar organic phase enabled the backward extraction of cytochrome c to be achieved without the formation of insoluble aggregates.

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