| 摘要 |
PURPOSE: A method for purifying a polygalacturonase is provided of which the steps are simplified. Thereby, the purified polygalacturonase, for example, Kluyveromyces marxianus IFO 0288, is provided. The purification method allows to design a primer by sequencing N-terminal amino acids-based sequences and increases an availability of the polygalacturonase. CONSTITUTION: Polygalacturonase is purified by the steps of: applying a concentrated culture medium containing a microorganism that secretes the polygalacturonase, to an ion-exchange column using 10-100 mM of sodium acetate (pH 5.0) as buffer and 10-100 mM of sodium acetate with a linear gradient of 0-0.2 M NaCl as an eluate to carry out a fast protein liquid chromatography(FPLC). The ion-exchange column is preferably mono S ion exchange column and the concentrated culture medium is obtained by a membrane filter thickener for centrifugation. Also, the microorganism is preferably Kluyveromyces marxianus IFO 0288 and the polygalacturonase has preferably a sequence set forth in SEQ ID NO : 1 at N-terminal thereof.
|
