一株纤维素降解菌的分离、鉴定及发酵优化  

Isolation,identification and fermentation condition optimization of a cellulose degrading strain

作　　者：李文兵 毕江涛[2] 惠治兵 刘鹏 王兵 孙权[1] LI Wenbing;BI Jiangtao;HUI Zhibing;LIU Peng;WANG Bing;SUN Quan(School of Agriculture,Ningxia University,Yinchuan 750021;School of Ecology and Environment,Ningxia University,Yinchuan 750021;Ningxia Yitaifeng Ecological Fertilizer Co.Ltd.,Yinchuan 750100)

机构地区：[1]宁夏大学农学院,银川750021 [2]宁夏大学生态环境学院,银川750021 [3]宁夏壹泰丰生态肥业有限公司,银川750100

出　　处：《安徽农业大学学报》2021年第3期458-466,共9页Journal of Anhui Agricultural University

基　　金：宁夏重点研发计划(2018ZDKJ0312)资助。

摘　　要：纤维素降解菌在堆肥发酵中具有加快腐熟、提升堆肥品质的作用,其中抗逆性强的芽孢杆菌属具有一定的优势。从新鲜牛粪中筛选具有降解粪污能力的纤维素降解菌,通过羧甲基纤维素钠培养基与革兰氏碘液结合培养、滤纸摇瓶复筛,并进行形态学观察、生理生化分析、16S rDNA基因序列对比鉴定,获得一株纤维素降解菌CK41,经NCBI提交序列BLAST比对后,鉴定为内生芽孢杆菌(Bacillus endophticus)(登录号:MT023630)。研究结果显示,利用单因素试验和响应面优化产酶条件试验,在温度为37℃,时间为72 h,pH为7.0,接种量为4%(V/V)时,获得羧甲基纤维素酶活量3.14 U·mL^(-1),较优化前提高了149.5%。结果表明,温度、时间和pH对于羧甲基纤维素酶活力是重要影响因素,接种量影响程度较小。菌株CK41具有较强的纤维素酶生产能力,在堆肥发酵中具有一定的潜力。Cellulose degrading bacteria can accelerate the maturation and improve the quality of compost during composting fermentation,especially Bacillus strain with strong stress resistance.In this study,we screened cellulose degrading bacteria with the ability to degrade manure from fresh cow dung,combined culture with Gram-iodine solution by sodium carboxymethyl cellulose medium,filter paper shaking flask,and morphological observation,physiological and biochemical analysis;we compared and identified 16S rDNA gene sequence and obtained a cellulose-degrading bacteria CK41,and identified it as endophytic bacillus(registration number:MT023630)after NCBI submission sequence alignment.The results showed that the carboxymethyl cellulase activity was 3.14 U·mL^(-1) after the strain fermentation at 37℃for 72 h in the medium of pH 7.0 with the inoculation amount of 4%(V/V),which was 149.5%higher than before the optimization.The result indicated that temperature,time and pH value were important factors affecting carboxymethyl cellulase activity,while the inoculation amount had little effect.In conclusion,strain CK41 has strong cellulase production capacity and certain potential in composting fermentation.

关 键 词：堆肥 纤维素降解菌 内生芽孢杆菌 分离 鉴定 响应面优化 

分 类 号：X172[环境科学与工程—环境科学]