Purification And Characterization of Mannanase Enzyme from Lentinula edodes Isolate and Evaluation of Its Activity in Removing Blood Stains from Cotton Fabrics.

Abstract

Shiitake mushroom is the Japanese common name for the genus Lentinula edodes, and Shiitake is a fungus of medicinal and industrial importance. The enzyme Mannanase was purified from the fungal culture filtrate. It included a series of steps, including precipitation with ammonium sulfate and a saturation rate of 80%, in which the specific activity reached 2 units/mg, and the enzymatic accumulation was 80%. It was followed by enzyme dialysis for a whole day, which caused an increase in the specific activity to 3 units/mg, the number of purification times reached 3 times, and the enzymatic accumulation reached 71%. It is noticed from the ion exchange step using DEAE-cellulose that the specific activity increased to 10 units/mg, the number of purification times reached 10 times, and the enzymatic accumulation reached 53%. Subsequently, the last purification step was carried out using gel filtration chromatography Sephadex G-150, in which the specific activity of the enzyme reached 24 units/mg and the number of purification times reached 24 times, with an enzymatic accumulation of 48%. The molecular weight was determined using electrophoresis in a polyacrylamide gel, and its molecular weight was 58 kilodaltons. The optimum pH for its activity was 6.5, while the pH ranged between (5.5-7) to prove its activity. The optimum temperature for enzyme activity was 60°C, while the thermal stability was between (20-80) °C. Keywords: Lentinula edodes, β-mannanase activity, purifi-cation, DEAE-Sephadex A-150, Characterization, removing and ability of detergents to clean clothes.