食品科学 ›› 2013, Vol. 34 ›› Issue (9): 175-179.doi: 10.7506/spkx1002-6630-201309036

• 生物工程 • 上一篇    下一篇

草酸青霉果胶酶分离纯化工艺及酶学性质研究

张名爱1,2,王宝维1,2,*,岳 斌2,荆丽珍2,3,葛文华2   

  1. 1.青岛农业大学食品科学与工程学院,山东 青岛 266109;
    2.青岛农业大学优质水禽研究所,山东 青岛 266109;3.海阳市畜牧兽医站,山东 烟台 265100
  • 收稿日期:2012-08-01 修回日期:2013-03-19 出版日期:2013-05-15 发布日期:2013-05-07
  • 通讯作者: 王宝维 E-mail:wangbw@qau.edu.cn
  • 基金资助:

    国家现代水禽产业技术体系建设项目(CARS-43-11);山东省农业科技成果转化基金项目(6621236)

Purification and Enzyme Properties of Pectinase from Penicillium oxalicu

ZHANG Ming-ai1,2,WANG Bao-wei1,2,*,YUE Bin2,JING Li-zhen2,3,GE Wen-hua2   

  1. 1. College of Food Science and Engineering, Qingdao Agricultural University, Qingdao 266109, China;
    2. Waterfowl Research Institute, Qingdao Agricultural University, Qingdao 266109, China;
    3. Haiyang Animal Husbandry and Veterinary Station, Yantai 265100, China
  • Received:2012-08-01 Revised:2013-03-19 Online:2013-05-15 Published:2013-05-07
  • Contact: WANG Bao-wei E-mail:wangbw@qau.edu.cn

摘要:

为了探索从鹅肠道分离的草酸青霉(Penicillium oxalicum)发酵生产果胶酶分离纯化新工艺,通过硫酸铵分级盐析、Sephacryl S-200分子筛柱层析、DEAE-Sephadex A-50离子交换层析技术对其发酵产物进行分离纯化,得到果胶酶的主要组分半乳糖醛酸酶(PG)和果胶酯酶(PE),并分别对PG和PE的最适作用pH值、温度以及分子质量等酶学特性进行研究。结果表明:PG和PE纯化倍数分别为32.21和23.98,均达到电泳纯,分子质量分别为61.4kD和42.2kD。PG最适作用pH值为5.0,最适反应温度为40℃,pH值稳定范围为4.0~6.0,在30、40℃随着处理时间延长酶活力保持稳定。PE最适作用pH值为6.0,最适温度为50℃,pH值稳定范围为5.0~7.0;PE在30℃时的热稳定性较好,40℃时处理60min,酶活力变化不明显。

关键词: 草酸青霉, 果胶酶, 分离纯化, 工艺, 酶学性质

Abstract:

Penicillium oxalicum separated from intestine of Wulong goose was used as fermentation microbe. Two major
components (PG and PE) of pectinase were purified from fermented products through salting out by ammonium sulfate,
Sephacryl S-200 gel filtration, and DEAE-Sephadex A-50 ion-exchange chromatography. The enzyme properties such as
optimal pH, temperature, and molecular weight were also studied. The results showed that the purification folds of PG and
PE were 32.21 and 23.98, respectively, and both of them reached up to electrophoretic purity and the molecular weights
of PG and PE were 61.4 kD and 42.2 kD, respectively. The optimal pH and temperature of PG was 5.0 and 40 ℃. PG was
stable in the range of pH 4.0-6.0. The enzyme activity remained stable between 30-40 ℃ with the extension of process
time. The optimal pH and temperature of PE was 6.0 and 50 ℃. PG was stable in the range of pH 5.0-7.0. The enzyme
activity was stable at 30 ℃ and the enzyme activity did not obvious change during 60 min treatment at 40 ℃.

Key words: Penicillium oxalicum, pectinase, separation and purification, technology, enzyme properties

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