Purification and Characterization of Lipase from the Liver of Carp, Cyprinus carpio L. (1758), Living in Lake Tödürge (Sivas, TÜRKİYE)
Öz
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.
Purification and Characterization of Lipase from the Liver of Carp, Cyprinus carpio L. (1758), Living in Lake Tödürge (Sivas, TÜRKİYE)
Öz
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.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Bölüm | Makaleler |
| Yazarlar | |
| Yayımlanma Tarihi | 1 Nisan 2012 |
| Yayımlandığı Sayı | Yıl 2012 Cilt: 12 Sayı: 2 |