Assay of Urease Enzyme Activity (Enzymology Practical Protocol)
This is the Biochemistry Protocols section post. Here we are providing “Assay of Urease enzyme activity” protocol. before going to the protocol we need to know few points about “Urease Enzyme”.
What is Urease ?
Urease is a hydrolytic enzyme that catalyzes the Urea into Carbon dioxide and ammonia. The Enzyme Commission Number is 126.96.36.199. This reaction follows
(NH2)2CO + H2O →CO2 + 2NH3
In 1926, James B. Sumner, an assistant professor at Cornell University, showed that urease is a protein by examining its crystallized form. Sumner’s work was the first demonstration that a pure protein can function as an enzyme, and led eventually to the recognition that most enzymes are in fact proteins and the award of the Nobel Prize in chemistry to Sumner in 1946.
Special characteristic features of Urease Enzyme:
- An active site requiring nickel in jack-beans and several bacteria. However, in vitro activation also has been achieved with manganese and cobalt
- Molecular weight: 480 kDa or 545 kDa for jack- bean Urease (calculated mass from the amino acid sequence). 840 amino acids per molecule, of which 90 are cysteines.
- Optimum pH: 7.4
- Optimum Temperature: 60 degrees Celsius
- Enzymatic specificity: Urea and Hydroxyurea
- Inhibitors: Heavy metals (Pb−& Pb2+)
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