The isolation and partial characterization of chymotrypsinogen from the pancreas of the ostrich (Struthio camelus) |
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Authors: | N van der Westhuizen R J Naudé W Oelofsen |
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Affiliation: | Department of Biochemistry, University of Port Elizabeth, Republic of South Africa. |
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Abstract: | 1. Cationic chymotrypsinogen from the pancreas of the ostrich was purified to homogeneity by sulfuric acid extraction of pancrei, (NH4)2SO4 fractionation and SP-Sephadex C-50 and Sephadex G-100 chromatography. 2. The final preparation was homogeneous when subjected to SDS-PAGE, isoelectric focusing and sedimentation equilibrium centrifugation. The Mmin value obtained from amino acid analysis was 25,572 Da. A mean sedimentation coefficient of 2.575 S was obtained by sedimentation velocity centrifugation. 3. N-terminal analysis by dansylation showed an Ala residue which is the N-terminal of a neochymotrypsinogen. 4. The effects of pH, temperature and inhibitors (LBTI, PMSF, TPCK and DFP) on the chymotryptic activity were examined. A Km-value for ATEE as substrate was found to be 0.57 mM. |
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