Properties of Klebsiella phage P13 and associated exopolysaccharide depolymerase

文献类型： 外文期刊

第一作者： Liu Yang

作者： Liu Yang;Chai Zihan;Shang Anqi;Mou Haijin;Li Guiyang;Mo Zhaolan

作者机构：

关键词： Klebsiella;bacteriophage;exopolysaccharide;depolymerase;enzymatic hydrolysis

期刊名称：JOURNAL OF OCEAN UNIVERSITY OF CHINA （ 影响因子：0.913； 五年影响因子：1.012 ）

ISSN： 1672-5182

年卷期： 2014 年 13 卷 1 期

页码：

收录情况： SCI

摘要： The bacteriophage P13 that infects Klebsiella serotype K13 contains a heat-stable depolymerase capable of effective degradation of exopolysaccharide (EPS) produced by this microorganism. In this study, the titer of phage P13, initially 2.0 x 10(7) pfu mL(-1), was found increasing 20 min after infection and reached 5.0 x 109 pfu mL(-1) in 60 min. Accordingly, the enzyme activity of depolymerase approached the maximum 60 min after infection. Treatment at 70A degrees C for 30 min inactivated all the phage, but retained over 90% of the depolymerase activity. Addition of acetone into the crude phage lysate led to precipitation of the protein, with a marked increase in bacterial EPS degradation activity and a rapid drop in the titer of phage. After partial purification by acetone precipitation and ultrafiltration centrifugation, the enzyme was separated from the phage particles, showing two components with enzyme activity on Q-Sepharose Fast Flow. The soluble enzyme had an optimum degradation activity at 60A degrees C and pH 6.5. Transmission electron microscopy demonstrated that the phage P13 particles were spherical with a diameter of 50 nm and a short stumpy tail. It was a double-strand DNA virus consisting of a nucleic acid molecule of 45976 bp. This work provides an efficient purification operation including thermal treatment and ultrafiltration centrifugation, to dissociate depolymerase from phage particles. The characterization of phage P13 and associated EPS depolymerase is beneficial for further application of this enzyme.

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