ABSTRACT
This work is aimed to study the optimal conditions of extraction and development of novel purification process for α-Amylase from pericarp of Borassus flabellifer fruit. Optimization was done by Response Surface Methodology (RSM) using Box-Behnken design. The variables chosen for this purpose are pH (X1), temperature (X2, °C) and CaCl2 concentration (X3, ppm). A total of 9 runs were conducted, varying each factor while others were kept constant. The contribution of each factor was established giving raise to significant t-values and p-values. This revealed that the three variables are essential for activity of the enzyme. Based on the obtained optimal conditions from RSM studies, enzyme formation was successfully achieved with an activity of 28.8 U and concentration of 1.4 mg/ml for the 90% ammonium sulfate precipitated dialyzed enzyme sample. A novel affinity chromatographic process was developed for purification process and by this process the enzyme concentration was found to be 0.032mg/ml and activity was determined as 4.76 U.
ABSTRACT
Solid state fermentation (SSF) was carried out for the production of Lglutaminase by the fungal strain Trichoderma koningii using sesamum oil cake as the solid substrate. L-glutaminase has received significant attention in recent years owing to its potential applications in medicine as an anticancer agent, as an efficient anti-retroviral agent and as a biosensor. In food industries it is used as a flavor and aroma enhancing agent. The overall maximum yield of L-glutaminase (19.41 U/gds) was achieved with the optimized process parameters of initial moisture content 65%, initial pH of the medium 7.0, supplemented with Dmaltose (1.0% w/v) and malt extract (1.0% w/v), inoculated with 2ml of 6 day old fungal culture and incubated at 33°C for 5 days. Both physico-chemical and nutritional parameters had played a significant role in the production of the enzyme, L-glutaminase. The enzyme production was found to be growth associated with the growth of the fungal culture.