瘤胃4株纤维降解细菌的分离鉴定及其纤维降解特性  

Isolation and Identification of Four Strain Cellulolytic Bacteria in Rumen of Sheep and Cellulose Degradation Characteristics

作　　者：王卫云[1] 李大彪[1] 于永强[1] 邢媛媛[1] 李子健[1] WANG Wei-yun LI Da-biao YU Yong-qiang XING Yuan-yuan LI Zi-jian(College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China)

机构地区：[1]内蒙古农业大学动物科学学院,呼和浩特010018

出　　处：《畜牧兽医学报》2016年第11期2294-2300,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金项目(31201822);内蒙古自治区自然科学基金项目(2011BS0403)

摘　　要：作者从蒙古绵羊瘤胃内容物中筛选出四株纤维素降解细菌并进行鉴定,将其编号为H1、H2、N1、N2,对菌株生长特性和纤维素酶活力进行测定,以期为后续试验提供试验材料。通过生理生化特征、DNA序列分析对分离菌株进行鉴定,比浊法测定细菌生长曲线,用还原糖法测定四株细菌的滤纸酶活、羧甲基纤维素酶活、水杨苷酶活和微晶纤维素酶活力。结果表明:经鉴定,四株菌中H1、H2为黄色瘤胃球菌(Ruminococcus flavefaciens),N1、N2为链球菌(Streptococcus);生长曲线测定表明菌株H1生长延迟期为4h,H2、N1、N2均为3h,黄色瘤胃球菌最大菌体浓度出现在28h,链球菌为14h;四株细菌均具有良好的纤维降解特性,其中H2滤纸酶活力、羧甲基纤维素酶活力、微晶纤维素酶活力显著高于菌株H1、N1、N2(P<0.05),H1滤纸酶活力最低,为0.08μmol·(mL·min)-1。本试验利用hungate滚管法分离出4株具有纤维素酶活力的细菌,可以满足后续试验需要。Four cellulose-degrading rumen bacteria strains were isolated from Mongolia sheep.The four strains bacteria were identified and numbered as H1,H2,N1 and N2.The growth characteristics and cellulose activity were measured to provide experimental material for subsequent experiments.Bacterial morphological,physiological and biochemical characterization,DNA sequence homology analysis were used for identification of isolated strains.Growth curve of strains were measured by using spectrophotometer.Filter paper enzyme activity,CMCase activity,salicin activity and microcrystalline cellulose enzyme activity of four strains of cellulolytic bacteria were measured by using reducing sugar method.Two strains of bacteria H1,H2 were identified as Ruminococcus flavefaciens and N1,N2 were Streptococcus.The growth curves of four strains showed that lag phase of H1,H2,N1,N2 were 3-4hand the maximum cell concentration of H1,H2 appeared in 28 hand N1,N2 appeared in 14 h.Four strains of bacteria had high activity of filter paper enzyme.The activity of filter paper enzyme,CMCase and microcrystalline cellulose enzyme were significantly higher for H2 than for H1,N1 and N2（P0.05）.Filter paper enzyme activity of H1 was 0.08μmol·（mL·min）-1 and was lowest among four strains.Four strains bacteria with cellulose enzyme activity were isolated by using the method of roll tube tech-nique.These four cellulolytic bacteria strains can be used for subsequent study.

关 键 词：瘤胃纤维素降解细菌 筛选 鉴定 纤维素酶 

分 类 号：S826[农业科学—畜牧学]