Digestive enzyme patterns and evaluation of protease classes in Catla catla (family: Cyprinidae) during early developmental stages.
Digestive enzymes of Catla catla were studied during ontogenic development. Specific amylase activity was 0.12+/-0.01 mg maltose mg protein(-1) h(-1) in fish 4 days after hatching (DAH) and reached a maximum on (0.41+/-0.12 mg maltose mg protein(-1) h(-1)) 34 DAH. Total protease activity was minimum (123.2+/-16.5 mU mg protein(-1) min(-1)) on day-8 and reached its highest level (2713+/-147.2 mU mg protein(-1) min(-1)) on day-32. Trypsin activity showed constant increasing trend from day-16 onwards and was maximum on day-34 (118.1+/-7.09 mU mg protein(-1) min(-1)). Highest chymotrypsin activity was found on day-32 (1789.0+/-111.7 mU mg protein(-1) min(-1)). Lipase activity was detected in 4 DAH catla. Lipase activity increased steadily from day-22 onwards. SDS-PAGE of crude enzyme extracts showed that high molecular mass bands (41.8-127.8 kDa) appeared during the early stages followed by low molecular mass bands (17.8-37.2 kDa). The number of protease activity bands in substrate SDS-PAGE increased with age of fish. During ontogenesis of carp, soybean trypsin inhibitor (SBTI), PMSF and TLCK inhibited 75.5+/-1.19% to 92.8+/-0.85%, 53.3+/-9.47% to 90.5+/-2.6% and 39.8+/-3.8% to 84.7+/-1.54% of total protease activity, respectively. There was only 2.58+/-0.66% to 10.21+/-0.09% inhibition of protease activity with EDTA. SBTI and PMSF inhibited 8 and 4 activity bands, respectively. TLCK, a specific trypsin inhibitor, inhibited four trypsin-like enzymes in carp during ontogenesis.
Rathore
R M
RM
Aqua Research Lab, Department of Zoology, University of Delhi, Delhi-110007, India.
Kumar
S
S
Chakrabarti
R
R
D000681:Amylases / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002918:Chymotrypsin / Q000737:chemistry; D003530:Cyprinidae / Q000196:embryology*; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D008049:Lipase / Q000378:metabolism; D008320:Maltose / Q000737:chemistry; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000737:chemistry / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity; D013997:Time Factors; D014105:Tosyl Compounds / Q000378:metabolism; D014357:Trypsin / Q000737:chemistry; D014360:Trypsin Inhibitor, Kunitz Soybean / Q000235:genetics
D011480:Protease Inhibitors; D014105:Tosyl Compounds; C003614:4-toluenesulfonyl fluoride; D008320:Maltose; D014360:Trypsin Inhibitor, Kunitz Soybean; D008049:Lipase; D000681:Amylases; D010447:Peptide Hydrolases; D002918:Chymotrypsin; D014357:Trypsin