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Turkish Journal of Fisheries and Aquatic Sciences 2012, Vol 12, Num, 2     (Pages: 207-215)

Purification and Characterization of Lipase from the Liver of Carp, Cyprinus carpio L. (1758), Living in Lake Tödürge (Sivas, Türkiye)

Salih Görgün 1 ,Mehmet Ali Akpınar 1

1 Cumhuriyet University, Science Faculty, Department of Biology, 58140 Sivas, Türkiye DOI : 10.4194/1303-2712-v12_2_03 Viewed : 4495 - Downloaded : 5190 In this study, a lipase from the liver of Cyprinus carpio Linnaeus (1758) living in Tödürge Lake (Sivas) was purified with purification parameters of 90.38 μmol/dk.mg protein specific activity and 75.50 fold purity. The purification procedure consisted of preparation of homogenate, precipitation with polyethylene glycol-6000 (PEG-6000) and chromatographic techniques including in Q sepharose, sephacryl S 200 HR and phenyl sepharose CL-4B, respectively. The purified lipase showed a single band with approximately a molecular weight of 74 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). pH and temperature optimums for the purified enzyme were determined as 8.0 and 37°C using p-nitrophenyl butirate (p-NPB) as substrate. Km and Vmax values were found as 0.17 mM p-NPB and 2.6 μmol/ml.dk, respectively. It was observed that surface active agents, such as sodium dodecyl sulfate (SDS), Triton X-100 and Nataurocholate act as inhibitor on lipase activity. Keywords : Cyprinus carpio, lipase activity, Q sepharose, sephacryl S 200 HR, phenyl sepharose CL-4B, Tödürge Lake