Isolation and properties of L-amino acid oxidase from Ophiophagus hannah venom.

Abstract

This paper reports the isolation and some properties of L-amino acid oxidase from Ophiophagus hannah venom. The differences between L-amino acid oxidase from Ophiophagus hannah and that from other sources in specific activity, properties and the spectrum of isozymes are noticeable. The result of electrophoresis on polyacrylamide gel shows that this purified enzyme is homogenous. The molecular weight determined by gradient polyacrylamide gel slab (4--30%) is around 15 X 10(4) dalton. The molecular weight of the subunit determined by SDS gradient gel electrophoresis (4--30%) is around 7.3 X 10(4) dalton. Therefore, this enzyme is composed of two subunits. The absorption spectrum reveals that there are two FADs in each molecule. The optimum pH of enzymic reaction is around 8.7--9.0 when L-leucine is used as substrate. The inhibition of the products is noticeable when substrate concentration goes beyond 3mM. This enzyme is heat stable and its activity would not decrease obviously after heating at 55 degrees C for 40 min. A linear relationship between enzyme concentration and reaction rate was noticed when enzyme concentration was below 5.7 micrograms/ml.