The isolation and partial characterization of chymotrypsinogen from the pancreas of the ostrich (Struthio camelus).
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.
van der Westhuizen
N
N
Department of Biochemistry, University of Port Elizabeth, Republic of South Africa.
Naudé
R J
RJ
Oelofsen
W
W
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001717:Birds / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D002852:Chromatography, Ion Exchange; D002919:Chymotrypsinogen / Q000302:isolation & purification* / Q000378:metabolism; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D013696:Temperature
D000596:Amino Acids; D002919:Chymotrypsinogen