Trypsin from Greenland cod, Gadus ogac. Isolation and comparative properties.

Area of Science:

• Biochemistry
• Enzymology
• Marine Biology

Background:

• Trypsin is a crucial digestive enzyme found in many vertebrates.
• Understanding fish trypsin diversity aids in biochemical and biotechnological applications.
• The Greenland cod (Gadus morhua) is a commercially important fish species.

Purpose of the Study:

• To isolate and characterize trypsin from the pyloric ceca of Greenland cod.
• To compare the properties of Greenland cod trypsin with trypsins from other species.
• To assess the enzyme's stability and activity under various conditions.

Main Methods:

• Isolation and purification of trypsinogen from Greenland cod pyloric ceca.
• Enzymatic assays using synthetic substrates (N alpha-benzoyl-DL-arginine p-nitroanilide, tosyl arginine methyl ester) and protein.
• Characterization of enzyme kinetics, pH stability, thermal stability, and molecular weight.
• Inhibition studies using phenylmethylsulfonyl fluoride (PMSF) and other trypsin inhibitors.

Main Results:

• Greenland cod trypsin was successfully isolated and demonstrated proteolytic activity.
• The enzyme was inhibited by PMSF and other known trypsin inhibitors, confirming its serine protease nature.
• It exhibited greater stability at alkaline pH compared to acidic pH and was sensitive to heat.
• Molecular weight was determined to be 23,500, with notable differences in disulfide bonds and alpha-helix content compared to other trypsins.
• Enzyme activity showed low responsiveness to temperature variations, maintaining high specific activity at lower temperatures.

Conclusions:

• Greenland cod trypsin is a serine protease with distinct biochemical properties.
• Its unique characteristics, including thermal stability and activity at low temperatures, may have specific ecological or biotechnological relevance.
• Further research could explore its potential applications in various industries.