Studies On Purification And Enzyme Characteristics Of Cellulase And Xylanase

At present,the spent mushroom compost enormous resources which had not been effectively utilized,caused adverse environment and a great waste of organic resources.The spent mushroom compost contained large amounts of residual extracellular enzymes,which had very wide industrial use value and application prospect.Hypsizygus marmoreus and Enoki mushroom is the main varieties in the factory production,providing a stable source of raw materials extraction in factory.lt has important significance to solve the energy shortage and environmental pollution through scientific and effective method in China.On the basis of single-factor test and orthogonal experimental design test,the remanent cellulase in the spent mushroom compost of Hypsizygus marmoreus and Xylase in the spent mushroom compost of Enoki mushroom were isolated and purified using ammonium sulfate salting-out,Sephadex G-100 column chromatography and DEAE C-52 anion chromatography exchange,and there enzymatic properties were investigated.For cellulase,experimental results showed that it exhibited a single band by the SDS-PAGE,and the molecular weight was estimated as 27.8KDa.The purification made the specific activity as high as 1121.82U/mg and purification multiple reached 12.13.The optimal temperature and pH value for activity were 50? and 4.8 respectively,and had a certain acid and alkali resistance.Cellulase was activated by Na+ and K+ and promoted by Mg2+ and Mn2+,wherease it was inhibited by Zn2+,Cu2+and Fe3+.The Vmax and Km values of the CMCase for CMC-Na were 24.45?g/mL/min and 9.77mg/mL,respectively.For xylase,resluts showed that it exhibited a single band by the SDS-PAGE,and the molecular weight was estimated as 37.8KDa.The purification made the specific activity as high as 946.67U/mg and purification multiple reached 12.74.The optimal temperature and pH value for activity were 50? and 6.0 respectively,and had a certain acid and alkali resistance and high-temperature-resisting.Xylanase was activated by Fe2+ and Zn2+ and promoted by Mn2+,K+ and Ca2+,wherease it was inhibited by Mg2+,Cu2+ and Fe3+.The Vmax and Km values of the Xylanase for xylan were 15.43 ?g/mL/min and 9.61 mg/mL,respectively.The research suggests that Xylanase has a good potential to be a cold-stable in paper manufacturing and feed industry.