生物技术通报 ›› 2020, Vol. 36 ›› Issue (2): 65-70.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0789

• 消化道微生物专题 • 上一篇    下一篇

小龙虾肠道产纤维素酶细菌的分离与鉴定

冯光志, 石慧, 刘博, 吴玉婷, 王月琳, 石玉   

  1. 武汉设计工程学院食品与生物科技学院,武汉 430205
  • 收稿日期:2019-08-29 出版日期:2020-02-26 发布日期:2020-02-23
  • 作者简介:冯光志,男,硕士,实验师,研究方向:应用微生物;E-mail:fenggz2004@126.com
  • 基金资助:
    湖北省教育厅优秀青年科技创新团队项目基金(T201534),武汉设计工程学院博士科研专项(k201711),国家级大学生科技创新项目(201814035003)

Screening and Identification of Cellulase-producing Strains Isolated from Crayfish Intestine

FENG Guang-zhi, SHI Hui, LIU Bo, WU Yu-ting, WANG Yue-lin, SHI Yu   

  1. College of Food and Biotechnology,Wuhan Institute of Design and Sciences,Wuhan 430205
  • Received:2019-08-29 Published:2020-02-26 Online:2020-02-23

摘要: 小龙虾肠道微生物是小龙虾降解纤维素和半纤维素的主要驱动力,挖掘小龙虾肠道内降解纤维素的益生菌资源及基因资源。采用纯培养法,从小龙虾肠道筛选产纤维素酶细菌,对高活性菌株进行高通量测序。形态学结合16S rRNA分子鉴定结果显示:菌株13为贝莱斯芽孢杆菌(Bacillus velezensis),菌株33为枯草芽孢杆菌(Bacillus subtilis),菌株33具有多种水解酶活性,并有较高的纤维素降解能力,高通量测序结果表明该菌基因组含有许多碳水化合物相关酶基因。小龙虾肠道中含有丰富的纤维素降解菌,芽孢杆菌属细菌占据一定比重。

关键词: 小龙虾肠道, 内生细菌, 纤维素酶, 分离鉴定, 高通量测序

Abstract: The intestinal microbes are the main driving force for crayfish to degrade cellulose and hemicellulose. In order to explore the probiotic resources and genetic resources of degrading cellulose in the crayfish intestine,the cellulase-producing bacteria were isolated from crayfish intestine by plate screening method,and the genome of the maximum active strain was analyzed by Illumina MiSeq high-throughput sequencing. Morphological and 16S rRNA gene molecular identification showed that strain 13 was Bacillus velezensis,and strain 33 was Bacillus subtilis. Strain 33 has multiple hydrolase activities and high cellulose-degrading ability. The high-throughput sequencing results indicated that the bacterial genome contained many carbohydrate-related enzyme genes. Crayfish intestines are rich in cellulose-degrading bacteria,and Bacillus bacteria accounts for a certain proportion.

Key words: crayfish intestine, endophytic bacterium, cellulase, screening and identification, high-throughput sequencing