Studies Of The Ramie Deguming Key Enzymes From Bacillus Subtilis 16A-hemicellulase Purification And Degumming Efficient

Bast fibre plant is a natural spinning material,it must be deguramed befor spinng.There are some disadvantages to chemical degumming such as wasting energy polluting environment and so on.Biodegumming is more advancing and has a bright application prospect. At present, the technical key problem lies in how to reduce the residual gum furtherly.Many researches showed that increasing pectinase of degumming key enzyme cannot persistently enhance the degumming effect. So the paper is mainly to discuss the effect of hemicellulase .So we purify mannase and xylanase to research the synergy with pectinase.A strain of Bacillus subtilis 16A,which has a strong degumming activity of ramie,the optimum media of mannase konjak gum is 30g/L, peptone 9g/L, yeast extract 2g/L, KH₂PO₄ 5/L, MgSO₄.7H2O 0.025g/L and the optimum fermentation condition is initial pH8.0, fermentation period 72h, filling level 50ml, inoculum size 10mL.The optimum media of xylanase is 70% corn lister:30% wheat bran, peptone 6g/L, MgSO4 1.0g/L,Tween80 0.5g/L;the optimum fermentation condition is inistial pH8.0, fermentation period 60h, filling level 50ml, inoculum size 10mL . These two kinds hemicellulase were purified according to the above culture medium composition and condition. Crude enzymes were treated in turn with ammonium sulfate salting out, dialysis, DEAE-Sepharose Fast Flow anion exchange chromatograph, Sephadex gel filtration chromatography, and finally SDS-PAGE discontinuous flow electrophoresis detected a singular strip. The basic character of the both hemianalysis showed that, their optimal pH was 7.0.They stabilized in neutral conditions (pH7.0-8.0) and easily lost activity when tempreture was more than 55℃; Mn²⁺, Mg²⁺, Al³⁺ were inhibitors of hemicellulase. At last we carry about degumming test with purified key enzymes and reduce the residual gum content to 5.62%.It lays the foundation of efficient enzymatic preparation.