华北地区圈养大熊猫粪便中产纤维素酶芽孢杆菌菌株的分离与鉴定

doi:10.3969/j.issn.1000-1565.2016.06.010

摘要/Abstract

摘要： 为分离筛选大熊猫肠道中具有纤维素降解能力的芽孢杆菌菌株,首先对8份大熊猫粪便进行高温水浴处理,利用刚果红平板法进行初筛和复筛,DNS法测定酶活力.结果分离出126株菌株,其中8株菌株纤维素降解能力较强.结合形态特征、生理生化特征和16S rDNA序列分析,鉴定8个菌株均属于芽孢杆菌属,其中1株枯草芽孢杆菌(Bacillus subtilis),1株阿萨尔基芽孢杆菌(B.axarquienis),2株西姆芽孢杆菌(B.siamensis),4株甲基营养型芽孢杆菌(B.methylotrophicus).实验表明大熊猫肠道中存在多种降解纤维素的芽孢杆菌,其中新发现的有大熊猫源的阿萨尔基芽孢杆菌和西姆芽孢杆菌,丰富了大熊猫肠道菌群的研究,并为高效生产纤维素酶提供新的菌种来源.探究了大熊猫消化道内高效的纤维素降解菌,有利于实现高纤维类饲料资源化利用.

关键词: 大熊猫, 粪便, 芽孢杆菌, 纤维素酶

Abstract: This study aimed at screening strains of Bacillus sp.capable of degrading cellulose and identifying the species of purpose strains.8 dung samples of giant panda were treated in water bath at high temperature.Congo red decolorizing ring method was used to screen the strains before DNS method to test the highest enzyme activity.The results showed that 8 strains of 126 strains isolated in dung of giant panda had highly efficient cellulose-degrading capability.Through morphological,physiological and biochemical analysis and the sequence analysis of 16S rDNA gene,8 strains were identified.The strains are all Bacillus sp.,including 1 Bacillus subtilis(2N-14),1 B.axarquienis(2N-12),2 B.siamensis(Y-6、Y-10),and 4 B.methylotrophicus(2N-10、2L-9、2L-24、3X-10).The test shows that there were many strains(Bacillus sp.)capable of cellulose degradation in the dung of giant panda.The giant panda-derived B.axarquienis,B.siamensis were isolated and cultivated for the first time,which enriched the research of giant panda intestinal flora and provided new sources of strains for efficient production of cellulose. Exploring the efficiency of- 2016年河北大学学报(自然科学版) 2016第36卷第6期 Journal of Hebei University(Natural Science Edition) Vol.36 No.6DOI:10.3969/j.issn.1000-1565.2016.06.010华北地区圈养大熊猫粪便中产纤维素酶芽孢杆菌菌株的分离与鉴定武红敏,郭威,郭晓军,周贤,朱宝成(河北农业大学生命科学学院,河北保定 071001)摘要:为分离筛选大熊猫肠道中具有纤维素降解能力的芽孢杆菌菌株,首先对8份大熊猫粪便进行高温水浴处理,利用刚果红平板法进行初筛和复筛,DNS法测定酶活力.结果分离出126株菌株,其中8株菌株纤维素降解能力较强.结合形态特征、生理生化特征和16S rDNA序列分析,鉴定8个菌株均属于芽孢杆菌属,其中1株枯草芽孢杆菌(Bacillus subtilis),1株阿萨尔基芽孢杆菌(B.axarquienis),2株西姆芽孢杆菌(B.siamensis),4株甲基营养型芽孢杆菌(B.methylotrophicus).实验表明大熊猫肠道中存在多种降解纤维素的芽孢杆菌,其中新发现的有大熊猫源的阿萨尔基芽孢杆菌和西姆芽孢杆菌,丰富了大熊猫肠道菌群的研究,并为高效生产纤维素酶提供新的菌种来源.探究了大熊猫消化道内高效的纤维素降解菌,有利于实现高纤维类饲料资源化利用.关键词:大熊猫;粪便;芽孢杆菌;纤维素酶中图分类号:S858.9 文献标志码:A 文章编号:1000-1565(2016)06-0623-12Isolation and identification of Bacillus capable of degradingcellulose from the feces captive giant panda of in north ChinaWU Hongmin,GUO Wei,GUO Xiaojun,ZHOU Xian,ZHU Baocheng(College of Life Sciences,Agriculture University of Hebei,Baoding 071001,China)Abstract:This study aimed at screening strains of Bacillus sp.capable of degrading cellulose and identifying the species of purpose strains.8 dung samples of giant panda were treated in water bath at high temperature.Congo red decolorizing ring method was used to screen the strains before DNS method to test the highest enzyme activity.The results showed that 8 strains of 126 strains isolated in dung of giant panda had highly efficient cellulose-degrading capability.Through morphological,physiological and biochemical analysis and the sequence analysis of 16S rDNA gene,8 strains were identified.The strains are all Bacillus sp.,including 1 Bacillus subtilis(2N-14),1 B.axarquienis(2N-12),2 B.siamensis(Y-6、Y-10),and 4 B.methylotrophicus(2N-10、2L-9、2L-24、3X-10).The test shows that there were many strains(Bacillus sp.)capable of cellulose degradation in the dung of giant panda.The giant panda-derived B.axarquienis,B.siamensis were isolated and cultivated for the first time,which enriched the research of giant panda intestinal flora and provided new sources of strains for efficient production of cellulose. Exploring the efficiency of- 收稿日期:2015-12-22 基金项目:河北省科技计划项目(12236606);保定市科学研究与发展计划项目(15ZN017) 第一作者:武红敏(1990—),女,河北藁城人,河北农业大学在读硕士研究生.E-mail:873659445@qq.com 通信作者:朱宝成(1962—),男,河北沧州人,河北农业大学教授,博士生导师,主要从事农牧微生物应用技术研究. E-mail:zhu2222@126.com河北大学学报(自然科学版) 第36卷第6期武红敏等:华北地区圈养大熊猫粪便中产纤维素酶芽孢杆菌菌株的分离与鉴定cellulose degradation bacteria in the digestive tract of panda is beneficial for the resource utilization of high fiber feed.

Key words: giant panda, feces, bacillus, cellulose

中图分类号:

S858.9

武红敏,郭威,郭晓军,周贤,朱宝成. 华北地区圈养大熊猫粪便中产纤维素酶芽孢杆菌菌株的分离与鉴定[J]. 河北大学学报(自然科学版), 2016, 36(6): 623-634.

WU Hongmin,GUO Wei,GUO Xiaojun,ZHOU Xian,ZHU Baocheng. Isolation and identification of Bacillus capable of degrading cellulose from the feces captive giant panda of in north China[J]. Journal of Hebei University (Natural Science Edition), 2016, 36(6): 623-634.

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