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Extraction, purification, and characterization of the lipase from the viscera of porgy (Stenotomus chrysops)

Lipase, from porgy (Stenotomus chrysops) viscera, was purified by polyethylene glycol (PEG) 1000 precipitation, followed by dialysis and affinity chromatography on EAH-Sepharose 48. The digestive lipase from porgy showed seasonal variation in activity with high activity found in late summer and early fall compared with a spring sample. Polyethylene glycol (PEG) precipitation fraction was used to characterize this enzyme using p-nitrophenyl palmitate (pNPP) as substrate. Porgy lipase did not behave like a bile salt activated/depended lipase because it was able to hydrolyze pNPP without bile salt (e.g., sodium cholate). Porgy lipase was stable within the pH range of pH 6.0-10.0, with an optimum activity at pH 8.5. The enzyme was quite stable at temperatures below 40°C, but lost its activity rapidly at temperatures above 40°C. The optimum activity for hydrolysis pNPP was at 40°C, but the enzyme also demonstrated relatively high activity at temperatures below 40°C (i.e., 10-40°C) as well. Detergents, Triton X-100, Tween 40 and Tween 80, at final concentrations of 0.5 mM and 1 mM were found to have inhibitory effects on porgy digestive lipase activity. However, all three tested detergents appeared to increase the activity of porgy digestive lipase at elevated temperatures (i.e., 60-80°C).

Identiferoai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.111549
Date January 2008
CreatorsTian, Feng, 1980-
PublisherMcGill University
Source SetsLibrary and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada
LanguageEnglish
Detected LanguageEnglish
TypeElectronic Thesis or Dissertation
Formatapplication/pdf
CoverageMaster of Science (Department of Food Science and Agricultural Chemistry.)
RightsAll items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated.
Relationalephsysno: 003132363, proquestno: AAIMR66729, Theses scanned by UMI/ProQuest.

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