Separation,Purification and Characterization of Three Endo-polygalacturonases from a Newly Isolated Penicillum oxalicum

【摘要】 Three endo-polygalacturonases(endoPGs) from a newly isolated Penicillum oxalicum(CGMCC 0907) capable of utilizing waste biomass as growth substrate were separated and purified to homogeneity by ultra-filtration,affinity adsorption chromatography,CM-cellulose column chromatography,and Sephadex G-100 gel filtration chromatography with the overall yield of 64.5% from the crude enzyme.The specific activities and recovery rates of endoPG-1,endoPG-2 and endoPG-3 were 1120 U/mg and 21.6%,1350 U/mg and 25.9%,and 1560 U/mg and 17.0%,respectively.The three purified endoPGs had a close molecular weight to 41 kDa as estimated by SDS-PAGE.The optimum temperature and pH for the function of them were 65℃ and 5.0,55℃ and 5.0,50℃ and 5.5,respectively.Their pI and Km values were 5.9 and 0.78 mg/mL,6.0 and 1.2 mg/mL,and 6.1 and 2.0 mg/mL,respectively.更多还原

【Abstract】 Three endo-polygalacturonases(endoPGs) from a newly isolated Penicillum oxalicum(CGMCC 0907) capable of utilizing waste biomass as growth substrate were separated and purified to homogeneity by ultra-filtration,affinity adsorption chromatography,CM-cellulose column chromatography,and Sephadex G-100 gel filtration chromatography with the overall yield of 64.5% from the crude enzyme.The specific activities and recovery rates of endoPG-1,endoPG-2 and endoPG-3 were 1120 U/mg and 21.6%,1350 U/mg and 25.9%,and 1560 U/mg and 17.0%,respectively.The three purified endoPGs had a close molecular weight to 41 kDa as estimated by SDS-PAGE.The optimum temperature and pH for the function of them were 65℃ and 5.0,55℃ and 5.0,50℃ and 5.5,respectively.Their pI and Km values were 5.9 and 0.78 mg/mL,6.0 and 1.2 mg/mL,and 6.1 and 2.0 mg/mL,respectively.更多还原