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›› 2009, Vol. 9 ›› Issue (2): 242-249.

• 反应与分离 • 上一篇    下一篇

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

张春辉 李祖明 彭霞薇 贾月 张洪勋 白志辉   

  1. 北京联合大学师范学院 北京联合大学师范学院 中国科学院生态环境研究中心 中国科学院生态环境研究中心 中国科学院生态环境研究中心 中国科学院生态环境研究中心
  • 收稿日期:2008-11-24 修回日期:2009-01-23 出版日期:2009-04-20 发布日期:2009-04-20
  • 通讯作者: 李祖明

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

ZHANG Chun-hui LI Zu-ming PENG Xia-wei JIA Yue ZHANG Hong-xun BAI Zhi-hui   

  1. Teacher's College of Beijing Union University Teacher's College of Beijing Union University The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences
  • Received:2008-11-24 Revised:2009-01-23 Online:2009-04-20 Published:2009-04-20
  • Contact: LI Zu-ming

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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.

关键词: separation, characterization, Penicillum oxalicum, endo-polygalacturonase

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.

Key words: separation, characterization, Penicillum oxalicum, endo-polygalacturonase

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