《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (8): 2398-2407.doi: 10.16431/j.cnki.1671-7236.2017.08.026

• 预防兽医 • 上一篇    下一篇

猪肠道中产蛋白酶解淀粉芽孢杆菌的筛选、鉴定及其酶学性质的初步研究

宁豫昌, 张晓静, 乔宏兴   

  1. 河南牧业经济学院, 郑州 450046
  • 收稿日期:2017-03-03 出版日期:2017-08-20 发布日期:2017-08-18
  • 通讯作者: 乔宏兴 E-mail:zzmzqhx@163.com
  • 作者简介:宁豫昌(1979-),男,河南郑州人,硕士,讲师,研究方向:微生物学,E-mail:ningyuchang1979@163.com
  • 基金资助:

    河南省自然科学基金(162300410128);河南牧业经济学院科研创新团队项目(HNUAHE2015001)

Screening and Identification of Protease-producing Bacillus amyloliquefaciens from Pig's Intestines and the Preliminary Study of Its Enzymatic Properties

NING Yu-chang, ZHANG Xiao-jing, QIAO Hong-xing   

  1. Henan University of Animal Husbandry and Economy, Zhengzhou 450046, China
  • Received:2017-03-03 Online:2017-08-20 Published:2017-08-18

摘要:

试验从健康猪肠道中筛选得到一株高产蛋白酶的芽孢杆菌MY-6菌株,通过形态学观察、生理生化试验和糖醇发酵试验、16S rDNA序列扩增与比对、Biolog鉴定等方法鉴定该菌株为解淀粉芽孢杆菌。并研究了蛋白酶活力与细菌生活周期的动态关系,pH、温度、金属离子和蛋白酶抑制剂对蛋白酶活力的影响及蛋白酶对不同底物的水解性能。结果显示,MY-6菌株蛋白酶的合成与其芽孢形成同步,均开始于对数生长期的前期,该菌株所产蛋白酶的最适作用条件为:40℃、pH 7.0,Ca2+和Mn2+对蛋白酶活力有明显的促进作用,PMSF能够完全抑制该蛋白酶的活力,说明该蛋白酶属丝氨酸蛋白酶,蛋白酶对鱼粉蛋白、大豆蛋白、玉米蛋白、酪蛋白的酶解物TCA可溶性肽含量分别为0.83、0.67、0.58和0.38 μmol/g。结果提示,解淀粉芽孢杆菌MY-6菌株可作为改善蛋白质消化的益生性芽孢杆菌的初步选择。

关键词: 猪; 解淀粉芽孢杆菌; 蛋白酶; 酶学性质

Abstract:

A Bacillus strains,MY-6 strain which secreted high amount of protease was isolated from healthy pig's intestines,and it was identified as Bacillus amyloliquefaciens by the methods of morphological observation, biochemical tests,16S rDNA sequence amplification and comparison, biolog identification. In order to get an initial understanding of the enzymatic properties of its protease, the dynamic relationship between protease activity and bacterial life cycle, effects of pH,temperature,metal ions and inhibitors on protease activity,and the protease hydrolysis of different substrates were investigated. The results showed that,the protease synthesis of MY-6 strain was synchronized with its spore formation which began in the early stage of the exponential growth phase. The optimum conditions for the protease was 40℃ and pH 7.0,Ca2+ and Mn2+ had a intensive effects on the protease activity, PMSF could completely inhibit the activity of protease, indicating that the protease was a serine protease.The content of TCA soluble peptides for protein hydrolyzate fish protein, soybean protein, corn protein and casein was 0.83,0.67,0.58 and 0.38 μmol/g,respectively. In conclusion,Bacillus amyloliquefaciens MY-6 strain could be used as an initial selection of probiotic Bacillus to improve protein digestion.

Key words: pig; Bacillus amyloliquefaciens; protease; enzymatic properties

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