tailieunhanh - Partial purification of intestinal triglyceride lipase from Cyprinion macrostomus Heckel, 1843 and effect of Ph on enzyme activity
Intestinal triglyceride lipase (TG) of Cyprinion macrostomus after precipitation with 30% polyethylene glycol (PEG), was partially purified by hydrophobic interaction chromatography on phenyl sepharose CL-4B, and the effect of pH on this enzyme activity was determined titrimetrically. Intestinal tissue homogenates were precipitated using 30% PEG-8000, and then applied onto a phenyl sepharose CL-4B column for hy-drophobic interaction chromatography. | Turk J Biol 26 (2002) 133-143 © TÜB‹TAK Partial Purification of Intestinal Triglyceride Lipase from Cyprinion macrostomus Heckel, 1843 and Effect of pH on Enzyme Activity* Naci DE⁄ERL‹ Cumhuriyet University, Science Faculty, Department of Biology, 58140, Sivas - TURKEY M. Ali AKPINAR Cumhuriyet University, Science Faculty, Department of Biology, 58140, Sivas - TURKEY Received: Abstract: Intestinal triglyceride lipase (TG) of Cyprinion macrostomus after precipitation with 30% polyethylene glycol (PEG), was partially purified by hydrophobic interaction chromatography on phenyl sepharose CL-4B, and the effect of pH on this enzyme activity was determined titrimetrically. Intestinal tissue homogenates were precipitated using 30% PEG-8000, and then applied onto a phenyl sepharose CL-4B column for hy-drophobic interaction chromatography. The lipase that bound to this hydrophobic resin and was partially purified by this single step application was then eluted from this resin with taurocholic acid and Triton X-100 elution. This purified enzyme effectively hydrolysed natural olive oil substrate under the experimental conditions. An optimal pH of , and yield, and purification parameters were estimated for this lipase. The molecular mass of this enzyme was determined to be 51 kDa under non-denaturing conditions by polyacrylamide disc-gel electrophoresis (Disc-PAGE). It was concluded that the intestinal lipase of this fish is alkaline in character. Key Words: Lipase, purification, enzyme activity, optimal pH. Cyprinion macrostomus HECKEL, 1843 ‹nceba¤›rsak Lipaz’›n›n K›smi Saflaflt›r›lmas› ve pH’n›n Enzim Aktivitesi Üzerine Etkisi Özet: Cyprinion macrostomus inceba¤›rsak Lipaz’›n›n% 30 polietilen glikol (PEG) ile çöktürülmesinden sonra, hidrofobik etkileflim kromatografisi ile k›smi saflaflt›r›lmas› ve pH’n›n enzimatik aktivite üzerine etkisi titrasyon yöntemiyle araflt›r›lm›flt›r. ‹nceba¤›rsak doku homojenat› %30 PEG-8000 ile çöktürülmüfl
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