Extraction and activity of chymotrypsin using AOT-DOLPA mixed reversed micellar systems

Masahiro Goto, Yoshio Ishikawa, Tsutomu Ono, Fumiyuki Nakashio, T. Alan Hatton

    研究成果: Contribution to journalArticle査読

    33 被引用数 (Scopus)

    抄録

    Novel reversed micellar solutions formulated with a mixture of AOT (dioctyl sulfosuccinate) and DOLPA (dioleyl phosphoric acid) show good potential for use in reversed micellar protein extraction operations. Chymotrypsin is easily extracted from an aqueous phase into organic isooctane containing 10 mM AOT and DOLPA in a 4:1 ratio. The extraction ability of the mixed reversed micelles of 10 mM was higher than that of 200 mM AOT alone. The results of extraction indicated that the AOT-DOLPA mixed reversed micelles are very useful for separating and enriching chymotrypsin. Back-extraction of chymotrypsin from the organic phase to a fresh aqueous phase is also accomplished by adding an alcohol to the organic phase. Although the backtransfer of chymotrypsin from the reversed micelles formed by AOT alone is very slow and difficult, in the AOT-DOLPA mixed reversed micelles, the back-extraction can be achieved completely by addition of 10% (v/v) isobutyl alcohol to the reversed micellar phase. The time to attain to the equilibrium of back-extraction was reduced from more than 24 to 2 h by adding the alcohol. On the basis of the activity data, the best composition of AOT and DOLPA was a 4:1 ratio and the total surfactant concentration was 10 mM. The activity of chymotrypsin recovered from the mixed reversed micelles was higher than that of the initial protein before the forward-transfer. This result means that the novel mixed reversed micellar solutions are useful not only in separation but also in purification of proteins.

    本文言語英語
    ページ(範囲)729-734
    ページ数6
    ジャーナルBiotechnology Progress
    14
    5
    DOI
    出版ステータス出版済み - 9 1 1998

    All Science Journal Classification (ASJC) codes

    • バイオテクノロジー

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