产纤维素酶枯草芽孢杆菌Kpg酶学性质研究及对麸皮纤维素含量的影响  

Enzyme Activity Analysis of Cellulase-producing Bacillus subtilis Kpg and its Effect on the Cellulose Content in the Bran

作　　者：聂利波 李旺[1] 史敦胜 宋洋洋[1] 王占彬[1] NIE Libo;LI Wang;SHI Dunsheng;SONG Yangyang;WANG Zhanbin(College of Animal Science and Technology,Henan University of Science and Technology,Luoyang,Henan 471003,China)

机构地区：[1]河南科技大学动物科技学院,河南洛阳471003

出　　处：《家畜生态学报》2018年第12期25-30,共6页Journal of Domestic Animal Ecology

基　　金：国家自然科学基金青年科学基金项目(31101744);河南省重大科技专项(131100110300)

摘　　要：对枯草芽孢杆菌B.subtilis Kpg所产纤维素酶的酶学性质进行研究,确定该纤维素酶的最佳温度和最适pH,并进行固体发酵麸皮试验,验证Kpg纤维素酶的实践效果。试验以3,5-二硝基水杨酸酶活测定法为基础,通过改变纤维素酶的反应温度和pH环境,确定最适反应温度和最适pH环境。为研究其实际降解能力,将枯草芽孢杆菌Kpg添加到麸皮中,在37℃环境中发酵6d。结果显示,枯草芽孢杆菌Kpg纤维素酶的最适反应温度为60℃,此时纤维素酶活性最大为108.45U/mL;反应液最适pH值为6.0,此时纤维素酶活性为97.72U/mL;在两者最适条件共同作用下,纤维素酶活性为113.58U/mL。发酵试验结果显示,枯草芽孢杆菌Kpg能够显著降低麸皮中的纤维素含量(P<0.05),同时显著提高发酵麸皮中的真蛋白含量(P<0.05)。枯草芽孢杆菌Kpg表现出的酶学性质符合发酵微生物选育的标准和特征,为生物饲料的开发和应用提供数据参考。In order to determine the optimum reaction temperature and pH,the enzymatic properties of cellulase which is produced by Bacillus subtilis Kpg is studied.And a solid fermentation bran test is designed to verify the practical effect of recombinant bacteria B.subtilis Kpg.In the experiment,the optimum reaction temperature and optimum pH are determined by changing the reaction temperature and pH of the cellulase.To study its actual ability to degrade cellulose,the recombinant bacteria B.subtilis Kpg is added to bran and fermented at 37℃for 6 d.The test results show that when the temperature of cellulase reaction is 60℃,the cellulase activity is the highest,reaching 108.45 U/mL at this temperature,and when the most suitable pH of cellulase reaction is 6,the cellulase activity is 97.72 U/mL at this pH.The enzyme activity of cellulase is 113.58 U/mL under conditions with the optimum temperature and pH coexisted.Fermentation test shows that the recombinant strain B.subtilis Kpg could decrease the content of cellulose in bran(P<0.05),and increase the content of true protein in bran(P<0.05).These enzyme activity of cellulase-producing B.subtilis Kpg meets the standards and requirements of a good fermentation microorganism,laying a theoretical basis for its development and application as a biological feed.

关 键 词：枯草芽孢杆菌 纤维素酶 酶活性分析 发酵 

分 类 号：S811.5[农业科学—畜牧学]