食品科学 ›› 2017, Vol. 38 ›› Issue (8): 117-121.doi: 10.7506/spkx1002-6630-201708019

• 生物工程 • 上一篇    下一篇

产耐热普鲁兰酶菌株的分离、筛选与鉴定

高 涛,王 芳,别小妹,吕凤霞,赵海珍,张 充,陆兆新   

  1. 南京农业大学食品科技学院,江苏 南京 210095
  • 收稿日期:2017-04-26 修回日期:2017-04-26 出版日期:2017-04-25 发布日期:2017-04-24

Isolation, Screening and Identification of a Bacterial Strain Producing Thermostable Pullulanase

GAO Tao, WANG Fang, BIE Xiaomei, Lü Fengxia, ZHAO Haizhen, ZHANG Chong, LU Zhaoxin   

  1. College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2017-04-26 Revised:2017-04-26 Online:2017-04-25 Published:2017-04-24

摘要: 从某火山口的土壤样品中分离筛选得到了一株产耐热普鲁兰酶菌株T-10,酶活力为1.22 U/mL。经形态特征观察、生理生化特征实验以及16S rDNA序列同源性分析,确定该菌株为解淀粉芽孢杆菌。酶学性质研究表明:该普鲁兰酶的最适温度是60 ℃,在70 ℃保温4 h后活力残留60%以上;最适pH值为6.0,pH 3.0~8.0在室温保存4 h后相对酶活力大约为70%。目前,鲜有发现产耐热普鲁兰酶的菌株为解淀粉芽孢杆菌,其具备良好的应用前景。

关键词: 普鲁兰酶, 筛选, 生理生化, 16S rDNA, 酶学性质

Abstract: themostable pullulanase-producing strain, designated as T-10, with enzymatic activity of 1.22 U/mL, was isolated from soil samples collected at a crater field. By applying morphological observation, physiological and biochemical tests and 16S rDNA sequence homology analysis, it was identified as Bacillus amyloliquefacienss. The optimum temperature for the pullulanase activity was 60 ℃ and more than 60% of its maximum activity was maintained after incubation at 70 ℃ for 4 h. The optimum pH value for the pullulanase activity was 6.0, and about 70% of its maximum activity was maintained in the range of 3.0?8.0 after incubation for 4 h. At present, only a few thermostable pullulanase-producing strains of B. amyloliquefaciens are available, and so this new strain of B. amyloliquefaciens has a good application prospect.

Key words: pullulanase, screening, physiological and biochemical characteristics, 16S rDNA, enzymatic properties

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