高通量筛选地衣芽孢杆菌DW2高产杆菌肽诱变菌株

doi:10.13560/j.cnki.biotech.bull.1985.2020-0080

生物技术通报 ›› 2020, Vol. 36 ›› Issue (7): 90-96.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0080

高通量筛选地衣芽孢杆菌DW2高产杆菌肽诱变菌株

米粱波¹, 曾伟主¹, 黄科学², 王得明², 堵国成¹, 周景文¹, 陈坚¹

1.江南大学生物工程学院,无锡 214122;
2.齐鲁制药（内蒙古）有限公司,呼和浩特 010080

收稿日期:2020-01-21 出版日期:2020-07-26 发布日期:2020-07-28
作者简介:米粱波,男,硕士研究生,研究方向：微生物的高通量筛选育种;E-mail：1107657607@qq.com
基金资助:
国家重点研究发展计划（2017YFC1600403）,国家自然科学基金优秀青年基金项目（21822806）,国家自然科学基金项目（31670095,31770097）,江苏省著名教授项目,中央高校基础研究基金（JUSRP51701A）,111项目（111-2-06）

High-throughput Screening High-yield Bacitracin Strain from Bacillus licheniformis DW2

MI Liang-bo¹, ZENG Wei-zhu¹, HUANG Ke-xue², WANG De-ming², DU Guo-cheng¹, ZHOU Jing-wen¹, CHEN Jian¹

1. School of Biotechnology,Jiangnan University,Wuxi 214122;
2. Qilu Pharmaceutical（Inner Mongolia）Co. ,Ltd. ,Huhhot 010080

Received:2020-01-21 Published:2020-07-26 Online:2020-07-28

摘要/Abstract

摘要： 杆菌肽是一种广泛使用的广谱多肽抗生素,其作为次级代谢产物具有复杂的生物合成过程。以杆菌肽工业生产菌株地衣芽孢杆菌DW2作为研究对象,采用ARTP诱变和基于流式细胞仪的高通量筛选策略,获取杆菌肽高产菌株。通过筛选确定了8#6D、7#5E、6#7E、5#8D和2#5F 5株高产菌株,相对出发菌株分别提高20.9%、20.9%、20.8%、19.7% 和22.2%,杆菌肽的最高效价达到912 U/mL。高通量筛选操作简单、安全,结合流式细胞仪高流速分选,可快速实现杆菌肽产量提高。

关键词: 杆菌肽, ARTP, 双缩脲反应, 流式细胞仪, 多孔微量滴定板

Abstract: Bacitracin,as secondary metabolite with complex biotransformation pathway,is a widely used broad-spectrum peptide antibiotic. An industrial strain Bacillus licheniformis DW2 was used for this study,and strains of high-yield bacitracin were obtained using the Atmospheric and Room Temperature Plasma（ARTP）mutagenesis and an established high-throughput screening strategy based on flow cytometry（FCM）. Finally,the 5 best performing strains with regard to bacitracin yield were determined by screening,and they were 8#6D,7#5E,6#7E,5#8D,and 2#5F. The yield increased by 20.9%,20.9%,20.8%,19.7%,and 22.2%,respectively,compared with that of starting DW2 strain. The highest titer of the bacitracin reached 912 U/mL. With the help of high-flow sorting by flow cytometry,high-throughput screening characterized of simple and safe can quickly increase bacitracin production.

Key words: bacitracin, ARTP, biuret coloration, flow cytometry, multi-well microtiter plates.

米粱波, 曾伟主, 黄科学, 王得明, 堵国成, 周景文, 陈坚. 高通量筛选地衣芽孢杆菌DW2高产杆菌肽诱变菌株[J]. 生物技术通报, 2020, 36(7): 90-96.

MI Liang-bo, ZENG Wei-zhu, HUANG Ke-xue, WANG De-ming, DU Guo-cheng, ZHOU Jing-wen, CHEN Jian. High-throughput Screening High-yield Bacitracin Strain from Bacillus licheniformis DW2[J]. Biotechnology Bulletin, 2020, 36(7): 90-96.

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高通量筛选地衣芽孢杆菌DW2高产杆菌肽诱变菌株

High-throughput Screening High-yield Bacitracin Strain from Bacillus licheniformis DW2

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