从黑曲霉发酵的果胶粗酶中分离纯化内切聚半乳糖醛酸酶.pdf

30 5 () Vol.30 No.5 2002 10 Journal of Fuzhou University(Natural Science) Oct.2002 :1000-2243(2002)05-0632-04 , , , , (, 　350002) :、CM-SephadexC-50 、POROSPE POROS HQ20 , .SDS 41.8 kD, pH 5.0 36℃. :;;; + :Q556 .2 :A Purification of an endo-polygalacturonase from Aspergillus niger LIN Hong-bao, FU Rong, XUE Wei-ming, GUO Chu -teng, RAO Ping-fa (Instituteof Biotechnology, Fuzhou University, Fuzhou, Fujia n350002, China) Abstract :A pectinasewas identified and isolated from a commercial Aspergillus niger pectinase prepa- ratio .The crude enzyme preparatio , which was prepared by precipitatio nof the water extract of the culture of Aspergillus niger with ammonium sulfate, wasfurtherfractionated by three stepsof chromatog- raphy of catio nexchange, hydrophobicinteractio nand anio nexchangetoobtai na nelectrophoretically ho- mogeneous pectinase.Themolecularweight of the purified enzymewas estimated by SDSto be 39.4 kD under no -reduced conditions and 41.8 kD under reduced conditions, with the optimum pH at 5.0, and the optimum temperature at 36 ℃. Keywords:Aspergillus niger ;endo-polygalacturonase;isolatio ;purificatio , 、、 [1] ., (EC3.2.1.15), , 、. , 、. 1 　 1.1 　 ();(5000 Millipore );CM-Seph- adex C-50(Pharmacia );POROS PE POROS 20 HQ (PE );(Sigma );();(Bio λ-10 , PE ); (270 Fractio nCollector, ChemLab );(Model AvantiTM J-251, Backma ); (126-solventmodule 166-detector, Backma );(Mini-protei nII cell, Bio-RAD );(HH ·S21·4 , ). 1.2 　 1.2 .1 　 4 20.0g 0.1 mol L 100 mL -(pH5.0), 4 ℃(1.0×10 r mi , 20 mi ), , 100mL.39.0g, 60%, 2mol LHCl :2