Development of A Method for the Stabilization and Formulation of Xylanase from Trichoderma Using Experimental Design

Abstract

The native purified Xylanase from Trichoderma is unstable when stored at elevated temperatures over time. To reduce the risk of activity loss associated with these storage temperatures, a series of designed experiments were conducted to determine the best method of enzyme stabilization. The formulation chemicals studied included polyols such as glycerol, sorbitol and propylene glycol with different salts and bactericidal components including sodium formate, potassium chloride, and sodium benzoate. Concentrations of these components were varied to yield maximum residual Xylanase activity in our stability experiments. Our studies show a maximum stability of endo 1,4-ß-D Xylanase produced from Trichoderma in the presence of glycerol and sodium formate in temperatures of 4°, 25°, 37° and 48°C, over an extended period of time. Sources of instability were identified through measurements of residual activity and visual clarity. The optimal liquid formulation was determined using statistical design experimentation in an accelerated study at 48°C to achieve a stable liquid Xylanase preparation. Xylanase activity was determined using the RBB-Azo-Birchwood insoluble substrate assay.