摘要
该研究将室温等离子(ARTP)诱变与微生物微滴培养(MMC)技术应用于几丁质脱乙酰基酶(CDA)高产菌株的诱变选育,构建高产CDA菌株的诱变及高通量筛选方法。结果表明,经过4轮的ARTP诱变及MMC筛选,从200个不同的液滴中共筛选出5个发酵产酶明显提升的液滴,并通过进一步的平板筛选、24-深孔板复筛,获得了17株产酶提高300%以上的诱变菌株。通过对比分析17株高产菌株产CDA的能力,确定了1株最佳CDA高产菌株B4,其CDA最大产量比出发菌株提高了3.15倍,发酵产酶总量达到419.11 U/mL,为原始菌种的3.90倍。该研究为CDA高产菌株的诱变选育及高通量筛选提供了借鉴。
In this paper,atmospheric and room temperature plasma(ARTP)mutagenesis and microbial microdroplet-culture(MMC)screening strategy were applied to the mutagenic breeding of strain with high chitin deacetylase(CDA)yield.The mutagenesis and high-throughput screening methods of high-yielding CDA strain were established.The results showed that a total of 5 droplets with significantly increased CDA fermentation were screened from 200 different droplets after four rounds of ARTP mutagenesis and MMC screening.17 mutagenesis strains with the CDA production increase of more than 300%were finally obtained through further plate screening,24-deep-hole plate rescreening verification.Finally,the optimal strain B4 with high yield of CDA was determined by comparing the CDA yield,and the maximum yield of CDA increased 3.15 times compared with that of the control.The study provided a reference for mutagenesis breeding and high-throughput screening of high-yielding CDA strains.
作者
马钦元
申雁冰
丁盼盼
屠琳娜
毕心宇
王敏
MA Qinyuan;SHEN Yanbing;DING Panpan;TU Linna;BI Xinyu;WANG Min(Key Laboratory of Industrial Fermentation Microbiology,College of Biotechnology,Tianjin University of Science and Technology,Tianjin 300457,China;Luoyang Tmaxtree Biotechnology Co.,Ltd.,Luoyang 471023,China)
出处
《中国酿造》
CAS
北大核心
2020年第8期170-174,共5页
China Brewing
基金
国家自然科学基金项目(21706194)
天津自然科学基金项目(17PTGCCX00190)。
关键词
几丁质脱乙酰基酶
选育
常压室温等离子诱变
微生物微滴培养
chitin deacetylase
breeding
atmospheric and room temperature plasma
microbial microdroplet-culture
