产纤维素酶细菌的筛选鉴定与特性分析

doi:10.11924/j.issn.1000-6850.casb2020-0855

中国农学通报 ›› 2021, Vol. 37 ›› Issue (30): 83-88.doi: 10.11924/j.issn.1000-6850.casb2020-0855

所属专题：生物技术

产纤维素酶细菌的筛选鉴定与特性分析

赵龙妹(), 陈林, 杜东晓, 董惠心, 李旺, 李元晓, 何万领, 曹平华

河南科技大学动物科技学院,河南洛阳 471000

收稿日期:2020-12-31 修回日期:2021-05-12 出版日期:2021-10-25 发布日期:2021-12-08
作者简介:赵龙妹,女,1988年出生,河南洛阳人,讲师,博士,研究方向：饲料生物技术与功能微生物开发。通信地址：471000 河南省洛阳市洛龙区开元大道263号河南科技大学动物科技学院,Tel：18638830711,E-mail： zhaolongmei@126.com。
基金资助:
河南省重点研发与推广专项(科技攻关)“酶解玉米芯新型饲料添加剂的开发”(212102110174);河南科技大学博士科研启动基金项目“酶解玉米芯与益生素联用效果研究”(13480076)

Screening, Identification and Characteristic Analysis of Cellulase-Producing Bacteria

Zhao Longmei(), Chen Lin, Du Dongxiao, Dong Huixin, Li Wang, Li Yuanxiao, He Wanling, Cao Pinghua

College of Animal Science and Technology, Henan University of Science and Technology, Luoyang Henan 471000

Received:2020-12-31 Revised:2021-05-12 Online:2021-10-25 Published:2021-12-08

摘要/Abstract

摘要：

为了筛选获得产纤维素酶细菌,为新型饲料添加剂的开发提供材料基础。本研究以玉米田土壤为样品,使用刚果红平板法筛选产纤维素酶细菌并分析其所产酶的特性。通过试验,筛选获得产纤维素酶菌株,经形态学和分子生物学法将其鉴定为巨大芽孢杆菌(Bacillus megaterium)。对巨大芽孢杆菌XT2所产纤维素酶进行酶学特性分析,发现其最适反应条件为50℃,pH 6.0,具有一定的热稳定性,K⁺对纤维素酶具有激活作用,Mg²⁺、Ca²⁺和Mn²⁺对酶活具有抑制作用。该菌生长至20 h时,所产纤维素酶活力最高,达到0.774 U/mL。该产纤维素酶芽孢杆菌的成功分离获取为饲料新资源的开发以及新型饲料添加剂的研制提供了菌种材料。

关键词: 纤维素酶, 芽孢杆菌, 筛选, 鉴定, 酶学特性

Abstract:

To screen cellulase-producing bacteria and provide a material basis for the development of new feed additives, in this study, soil samples from corn fields were used to screen cellulase-producing bacteria by congo red plate method, and the characteristics of cellulase produced by it were analyzed. Through the test, the cellulase-producing strain was screened and identified as Bacillus megaterium by morphological and molecular biological methods. The enzymatic characteristics of cellulase produced by Bacillus megaterium XT2 was analyzed and the optimal reaction condition was found to be 50℃ and pH 6.0, and the cellulase showed certain thermal stability, K⁺ could activate the cellulase activity, and Mg²⁺, Ca²⁺ and Mn²⁺ had an inhibitory effect on enzyme activity. When the strain grew to 20 h, the cellulase activity reached the highest (0.774 U/mL). The successful isolation of cellulase-producing Bacillus could provide a strain material for the development of new feed resources and new feed additives.

Key words: cellulase, Bacillus, screening, identification, enzymatic characteristics

中图分类号:

赵龙妹, 陈林, 杜东晓, 董惠心, 李旺, 李元晓, 何万领, 曹平华. 产纤维素酶细菌的筛选鉴定与特性分析[J]. 中国农学通报, 2021, 37(30): 83-88.

Zhao Longmei, Chen Lin, Du Dongxiao, Dong Huixin, Li Wang, Li Yuanxiao, He Wanling, Cao Pinghua. Screening, Identification and Characteristic Analysis of Cellulase-Producing Bacteria[J]. Chinese Agricultural Science Bulletin, 2021, 37(30): 83-88.

图/表 9

图1. 刚果红染色后菌落周围的透明圈

图2. 菌株XT2的形态学观察

图3. 菌株XT2的系统发育树

图4. 温度对酶活力的影响

图5. pH对酶活力的影响

图6. 纤维素酶的耐热性

图7. 纤维素酶的耐碱性

图8. 不同金属离子对纤维素酶活力的影响

图9. 菌株XT2的生长曲线和产酶规律

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产纤维素酶细菌的筛选鉴定与特性分析

Screening, Identification and Characteristic Analysis of Cellulase-Producing Bacteria

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