Isolation of a Complex of Endo-cellulases from the Gastric Teeth of Dolabella auricularia by Affinity Chromatography(Biological Chemistry)

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Cellooligomer-(EPOX)-Sepharose CL-2B affinity chromatography was used to further purify the main endo-cellulase fraction, E_3-II, obtained from the gastric teeth of the marine mollusc Dolabella auricularia. The cellulase component in E_3-II, which had a strong affinity for the adsorbent in the presence of 1.0M NaCl, was effectively removed from the non-cellulase protein and separated into two fractions, one (E_3-II-N) with a low affinity and the other (E_3-II-A) with a high affinity for the adsorbent. E_3-II-A, the main component of E_3-II, was desorbed with a buffer solution without NaCl or by the addition of 10mM cellotriitol to the eluent with or without 1.0M NaCl, and it was purified 23.1-fold over the starting material. Fraction A, obtained by further purification of E_3-II-A by gel filtration, was a single protein band on SDS-PAGE (M.W.: 51,000), and separated into seven protein bands on DISC-PAGE. At least six of the protein bands had individually been demonstrated to be the same molecular weight (M.W.: 31,000) by Hedrick-Smith plots on DISC-PAGE as by gel filtration. Fraction A was most active at pH 6.3 and it had a typical endo-hydrolysis pattern on CMC.
社団法人日本農芸化学会の論文
1988-03-23