食品科学 ›› 2011, Vol. 32 ›› Issue (21): 183-187.doi: 10.7506/spkx1002-6630-201121037

• 生物工程 • 上一篇    下一篇

高产蛋白酶细菌的分离筛选及其种类鉴定

马桂珍,暴增海,王淑芳,吴少杰,付泓润,葛平华   

  1. 淮海工学院海洋学院
  • 收稿日期:2018-06-20 修回日期:2018-06-20 出版日期:2011-11-15 发布日期:2011-11-11

Screening and Identification of High Protease-producing Bacteria

MA Gui-zhen,BAO Zeng-hai,WANG Shu-fang,WU Shao-jie,FU Hong-run,GE Ping-hua   

  1. (School of Marine Science and Technology, Huaihai Institute of Technology, Lianyungang 222005, China)
  • Received:2018-06-20 Revised:2018-06-20 Online:2011-11-15 Published:2011-11-11

摘要: 从广东、河北、辽宁、湖南、江苏等省份采集土壤样品9个,采用稀释涂布的方法共分离到细菌菌株44株,利用平板透明圈法对44株细菌菌株产蛋白酶能力进行初步筛选,16株菌株出现了明显的透明圈,菌株G1-b、GD-2-2、HX-B-1的蛋白酶活性最强,透明圈/菌落直径比(HC)在3.5以上;比色法测定产酶活性强的3株菌株的蛋白酶酶活力,GD-2-2菌株的产酶能力最强,酶活力达到447.6U/mL。通过对细菌GD-2-2菌株形态学观察、生理生化实验,结合相关文献,初步鉴定菌株GD-2-2属于芽孢杆菌属。对菌株GD-2-2的 16S rDNA序列分析表明,该菌株序列与细菌Bacillus licheniformis strain YB915 (GQ996726.1)的16S rDNA序列同源性达100%,综合形态学观察和生理生化实验结果,鉴定GD-2-2菌株为地衣芽孢杆菌(Bacillus licheniformis)。

关键词: 细菌, 蛋白酶, 鉴定

Abstract: Totally 44 bacterial strains were isolated from 9 soil samples collected from Guangdong, Hebei, Liaoning, Hunan and Jiangsu by diluted plate method. The protease-producing ability of the bacteria isolated was screened by transparent cycle plate assay. The results indicated that of these bacterial isolates, 16 exhibited clear transparent cycles. Meanwhile, strains G1-b, GD-2-2 and HX-B-1 revealed the strongest protease activity. The ratio of transparent circle diameter to colony diameter was higher than 3.5. Strain GD-2-2 had stronger protease activity than the other two, reaching 447.6 U/mL. Based on morphological observations and physiological and biochemical tests, strain GD-2-2 belonged to Bacillus. The 16S rDNA sequence analysis showed that the isolate was 100% similar with Bacillus licheniformis strain YB915 (GQ996726.1). Therefore, strain GD-2-2 belonged to Bacillus licheniformis.

Key words: bacteria, protease, identification

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