沼液中一株促沼气发酵的芽孢杆菌的分离与鉴定  

Isolation and Characterization of Bacillus flexus LAM-CQ-3,a Methanogenesis Facilitating Bacillus Isolated from Biogas Slurry

作　　者：刘超奇[1] 向廷生[1] 陈超[2] 王彦伟[3] 宋金龙[3] 庄严[3] 郝元福[3] 卢麦田[3] 吴进[4] 阮志勇[3] 

机构地区：[1]长江大学地球环境与水资源学院,湖北荆州434023 [2]东北农业大学资源与环境学院,黑龙江哈尔滨150030 [3]中国农业科学院农业资源与农业区划研究所,北京100081 [4]农业部沼气科学研究所,四川成都610041

出　　处：《中国沼气》2013年第3期8-11,28,共5页China Biogas

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(202-37);"十二五"农村领域国家科技计划课题(2011BAD11B05);现代牧业(集团公司)攻关项目(MF20100518);国家科技支撑计划(2010BAD03B01);石家庄市科学技术研究与发展计划(12849057A)

摘　　要：从现代牧业集团塞北牧场的规模化沼气工程沼液中分离得到10株芽孢杆菌,其中3株可以促进沼气产生,通过实验室模拟沼气发酵试验,发现其中LAM-CQ-3菌株可以明显促进沼气的产生,沼气产量比对照提高126.72%。通过形态特征、生理生化特征、(G+C)mol%含量及16S rRNA序列分析等鉴定该菌为弯曲芽孢杆菌(Bacillus flexus)。LAM-CQ-3的特征为菌体杆状,革兰氏染色阳性,最适生长温度为35℃,最适生长pH为7.5,最适生长盐浓度为5%,与Bacillus flexus IFO15715T的相似性为99.58%,(G+C)mol%含量为46.6 mol%,主要脂肪酸组成为anteisoC 15:0(30.35%),isoC 15:0(22.59%),anteisoC 17:0(16.74%),isoC 17:0(10.24%),C 16:0(5.20%)。以16S rDNA序列为基础构建了包括16株邻近种属细菌在内的系统发育树,其中与模式弯曲芽孢杆菌的同源性最近。10 strains belong to the genus Bacillus were isolated from biogas slurry of a largescale anaerobic digester on Sai- bei dairy farm of Mordern Farming Corporation in Hebei, among which 3 strains were proved to be able to facilitate metha- nogenesis according to the results of simulated anaerobic digestion experiment in laboratory, and strain LAM-CQ-3 was se- lected for further study. The strain LAM-CQ-3 was identified and characterized based on colony morphological, physio-bio- chemical characteristic and sequencing of 16S rRNA. The strain had optimum growth temperature of 35~C, optimum growth pH of 7.5 and optimum growth salinity of 5%. The genomic G ＋ C content was 46.6mo1%. The 16S rRNA gene sequence analysis revealed that strain LAM-CQ-3 was a member of the Genus Bacillus, and was most closely related to Bacillus flex- us, showing 99.58% sequence similarity. The major fatty acids of strain LAM-CQ-3 were anteisoC 15：0 （30. 35% ） ,isoC 15：0 （22.59%）, anteisoC 17：0 （ 16.74% ） ,isoC 17：0 （ 10.24% ） ,C 16：0 （5.20%）. On the basis of its phenotypic and genotypic properties, strain LAM-CQ-3 was identified as Bacillus flexus LAM-CQ-3. The simulated methanogenesis experiment results showed that strain LAM-CQ-3 may possess potential to be used in anaerobic digestion.

关 键 词：沼液 沼气发酵 鉴定 弯曲芽孢杆菌(Bacillus flexus) 

分 类 号：S216.4[农业科学—农业机械化工程]