Purification and partial characterization of ostrich skeletal muscle cathepsin D and its activity during meat maturation.
Cathepsin D was purified from ostrich (Struthio camelus) skeletal muscle using pepstatin-A chromatography. The enzyme was comprised of two subunits (29.1 and 14 kDa). The N-terminal amino acid sequence of both subunits were determined and showed high amino acid sequence identity to other cathepsin D homologs. Ostrich cathepsin D was optimally active at pH 4 and at a temperature of 45°C, and was strongly inhibited by pepstatin-A (K(i)=3.07×10(-9)M) and dithiothreitol. Cathepsin D activities from five ostriches were monitored over a 30-day period. Cathepsin D remained substantially active throughout the 30-day storage period with an average remaining activity of 112±8.57% at day 30 (mean value from 5 ostriches).
Department of Biochemistry and Microbiology, Nelson Mandela Metropolitan University, P.O. Box 7700, Port Elizabeth 6000, South Africa.
Jason
J
Krause
Shonisani C
SC
Tshidino
Tomohisa
T
Ogawa
Yasuharu
Y
Watanabe
Vaughan
V
Oosthuizen
Benesh
B
Somai
Koji
K
Muramoto
Ryno J
RJ
Naudé
D000595:Amino Acid Sequence; D000818:Animals; D030161:Avian Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D002402:Cathepsin D / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D002846:Chromatography, Affinity; D004795:Enzyme Stability; D005511:Food Handling; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008297:Male; D008460:Meat / Q000032:analysis*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009124:Muscle Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D018482:Muscle, Skeletal / Q000201:enzymology*; D010436:Pepstatins / Q000378:metabolism / Q000494:pharmacology; D011480:Protease Inhibitors / Q000378:metabolism / Q000494:pharmacology; D021122:Protein Subunits; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D020310:Struthioniformes / Q000378:metabolism*; D013696:Temperature
D030161:Avian Proteins; D009124:Muscle Proteins; D010436:Pepstatins; D011480:Protease Inhibitors; D021122:Protein Subunits; D002402:Cathepsin D; C031375:pepstatin