一株易转化、高生物量地衣芽孢杆菌的筛选与鉴定
Screening and Identification of a Bacillus licheniformis Strain with High Electro-transfection Efficiency and Elevated Biomass
杜薇 1李志敏 1邢晏铭 1刘蒲临 1缪礼鸿1
作者信息
- 1. 武汉轻工大学生命科学与技术学院,武汉 430023
- 折叠
摘要
[目的]地衣芽孢杆菌(Bacillus licheniformis)是发酵工业中表达异源蛋白的重要底盘细胞,筛选易转化和高生物量的B.licheniformis菌株,为有效提升宿主的改造效率和蛋白表达水平提供菌种资源.[方法]经过富集、gyrB扩增与 16S rDNA序列分析从土壤中筛选与地衣芽孢杆菌同源性较高的菌株,将所筛菌株的电转效率和生物量与工业常用底盘细胞B.licheniformis 2709 进行比较分析.对筛选出的易电转和高生物量菌株进行形态学与生理生化鉴定,并进行抗生素敏感性测定和高温碱性环境生长情况测定.[结果]筛选出 10 株与地衣芽孢杆菌同源性较高的菌株,其中菌株 1-33 电转化效率为 6700 CFU/μg DNA,是B.licheniformis 2709的 13.7 倍.在豆粕玉米粉半固体培养基和SR液体培养基中培养,菌株 1-33 最大生物量达到 5.2×1010 CFU/mL和 6.8 g/L,分别为B.licheniformis 2709 的 1.71 倍和 1.3 倍.进一步经过形态学、生理生化特性实验将菌株 1-33 最终鉴定为B.licheniformis,并表现出多种常用抗生素的敏感性和良好的耐碱与耐高温的能力.[结论]筛选出一株易电转和高生物量的B.licheniformis 1-33,为开发成高效表达外源蛋白的宿主菌奠定基础.
Abstract
[Objective]Bacillus licheniformis is an important chassis cell for expressing heterologous proteins in the fermentation industry.Screening B.licheniformis strains with easy electro-transfection and high biomass provides strain resources for effectively improving the host modification efficiency and protein expression level.[Method]Screening of strains with high homology to B.licheniformis from soil was carried out through enrichment,gyrB amplification,and 16S rDNA sequence analysis.The electro-transfection efficiency and biomass of the screened strains were compared and analyzed with the commonly used chassis cell B.licheniformis 2709 in the industry.The screened strains with easy electro-transfection and high biomass were identified morphologically and physiologically and biochemically as well as were subjected to antibiotic susceptibility assay and growth in high temperature and alkaline environment.[Result]Ten strains with high homology to B.licheniformis were screened.Among them,the strain 1-33 presented the highest electro-transfection efficiency with a maximum value of 6700 transformants per μg DNA,which was 13.7 times higher than that of B.licheniformis 2709.Cultured in semi-solid soymeal-corn flour medium and SR liquid medium,the maximum biomass of strain 1-33 was found to be 5.2×1010 CFU/mL and 6.8 g/L,respectively,surpassing B.licheniformis 2709 by 1.71 and 1.3-fold.Further morphological and physiological experiments identified strain 1-33 as Bacillus licheniformis,which had sensitivity to various commonly used antibiotics and good alkaline and heat resistance.[Conclusion]A B.licheniformis strain 1-33 with high electro-transfection efficiency and elevated biomass is screened,thereby foundation for its further development as an efficient host for exogenous protein expression in industrial process is established.
关键词
地衣芽孢杆菌/易转化/高生物量/筛选/鉴定Key words
Bacillus licheniformis/easy electro-transfection/high biomass/screening/identification引用本文复制引用
基金项目
国家重点研发计划(2021YFC2100202)
出版年
2024
国家科技期刊平台
NSTL
万方数据