摘要
【目的】筛选高产纤维素酶菌株,为牛粪快速无害化降解提供菌剂。【方法】采用平板稀释法从重庆三峡库区3个牛粪堆肥厂样品分离纯化菌株,利用水解圈法和羧甲基纤维素(Carboxymethyl cellulose, CMC)酶糖化力法筛选、测定菌株纤维素酶活性,通过菌株形态特征观察、生理生化及分子生物学方法鉴定菌株,采用分子生物学软件MEGA6.0对菌株16S rRNA基因进行遗传系统发育树构建。【结果】从牛粪堆肥样品中分离得到的57株菌株中筛选出10株有较高纤维素酶活性,其中菌株X10纤维素酶活最高,为31.9 U·mL-1。菌株X10鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens),其在16S rRNA基因遗传系统发育树中与解淀粉芽孢杆菌位于同一簇。【结论】筛选的菌株X10为高产纤维素酶解淀粉芽孢杆菌,可用于牛粪无害化降解。
【Objective】 To screen and isolate a cellulase-producing microbe for efficient, rapid and natural composting cattle waste. 【 Method】 At 3 cow manure composting plants in the Three Gorges Reservoir area in Chongqing, samples for microbial analysis were collected. The agar dilution method was applied to isolate bacillus strains from the compost specimens.Cellulase activity of the strains was determined for the screening by the hydrolytic circle method and the CMC enzyme saccharification method. Selected strains were morphologically examined and further identified by physiological, biochemical,and molecular biological methods. Based on 16 S rRNA sequence, phylogenetic tree of the final selection was constructed using the molecular biological software MEGA6.0 for confirmation. 【Result】 Out of 57 isolates from the compost specimens, 10 strains showed varying degrees of cellulase activity. Among them, X10 had the highest activity at 31.9 U·mL-1 and was identified on the 16 S rRNA phylogenetic tree to be Bacillus amyloliquefaciens. 【 Conclusion】 X10 was a species of B.amyloliquefaciens that was highly productive in secreting cellulase to enhance the degradation of cow manure in compost.
作者
何深宏
程方俊
罗干
张耕
杜亚楠
康霞梅
王晓涵
赵自亮
任绍科
郭建华
HE Shenhong;CHENG Fangjun;LUO Gan;ZHANG Geng;DU Yanan;KANG Xiamei;WANG Xiaohan;ZHAO Ziliang;REN Shaoke;GUO Jianhua(College of Animal Science,Southwest University,Chongqing 402460,China)
出处
《福建农业学报》
CAS
CSCD
北大核心
2020年第7期781-787,共7页
Fujian Journal of Agricultural Sciences
基金
重庆市社会民生科技创新专项(cstc2016shmszx80068)
关键词
牛粪发酵
纤维素酶
解淀粉芽孢杆菌
Cow manure compost
cellulase
Bacillus amyloliquefaciens
作者简介
何深宏(1996−),男,硕士研究生,主要从事兽医微生物研究(E-mail:2577437282@qq.com);通信作者:程方俊(1962−),男,博士,副教授,主要从事预防兽医学研究(E-mail:cfjxndx@163.com);通信作者:郭建华(1977−),男,博士,副教授,主要从事微生物研究(E-mail:guo0619@163.com)
