Optimizing the preparation conditions and characterization of cross-linked enzyme aggregates of a monoamine oxidase

Abstract

Monoamine oxidases are useful in determination of biogenic monoamines, particularly histamine and tyramine. In this study, cross-linked enzyme aggregates (CLEAs) technique was applied to improve the stability of a monoamine oxidase from Arthrobacter aurescens (AMAO). Under the optimized condition (50% of saturated ammonium sulfate, 5 mM glutaraldehyde, 2.0 mg/mL AMAO, 4 h-cross-linking at 25 degrees C, pH 8.0), CLEAs-AMAO was recovered with a yield of 82% based on the subjected total enzyme activity. Both pH activity and stability at alkaline pHs of CLEAs-AMAO were significantly improved compared to those of the free enzyme, resulting in the shift of optimum pH to pH 8.0 and a broader pH profile. The half-life of the CLEAs at 65 degrees C was elongated by 1.7-fold compared to that of the free enzyme, suggesting the thermal stability of AMAO was also improved by the CLEAs formation.