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Buffalo (Bos buffali L.) chymosin purification and properties
C A Abdel Malak1, I F Abou El Adab, V Vukashinovic
1Chemistry Department, Mansoura University, Egypt.
Summary
Buffalo chymosin, an enzyme similar to bovine chymosin, was isolated and characterized. Subtle structural and functional differences were noted, particularly in its optimal pH for hemoglobin activity.
Area of Science:
- Biochemistry
- Enzymology
- Proteomics
Background:
- Chymosin is a key enzyme in milk coagulation.
- Investigating homologous enzymes from different species aids in understanding structure-function relationships.
Purpose of the Study:
- To isolate and characterize buffalo chymosin.
- To compare its properties with bovine chymosin.
Main Methods:
- Affinity chromatography using gramicidin S-agarose.
- Ion exchange chromatography on gamma-aminopropylsilochrom.
- Molecular weight determination and N-terminal sequencing.
Main Results:
- Buffalo chymosin isolated with a molecular weight of 36 ± 1 kDa.
- Identical N-terminal sequence to bovine chymosin, but differences in amino acid composition.
- Similar proteolytic and milk-clotting activities, but a higher pH optimum (4.0) for hemoglobin activity compared to bovine chymosin.
Conclusions:
- Buffalo and bovine chymosin share similar structures and functions.
- Subtle differences exist, particularly in pH optima, indicating distinct functional properties.