文献类型：期刊文献

英文题名：Breeding of L-tryptophan high-yield strain by ARTP mutagenesis and high-throughput screening; [基于常压室温等离子体诱变和高通量筛选选育L-色氨酸高产菌株]

作者：Ye H.; Du J.; Tang Y.; Yuan X.; Zhang Z.; Li L.; Ji Y.

第一作者：Ye H.

机构：[1]Beijing Key Laboratory of Biomass Waste Resource Utilization, College of Biochemical Engineering, Beijing Union University, Beijing, 100023, China;[2]Research Institute, Beijing Tsingke Biotech Co., Ltd., Beijing, 100000, China;[3]Key Laboratory for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Center for Synthetic & Systems Biology, Institute of Biochemical Engineering, Tsinghua University, Beijing, 100084, China

第一机构：北京联合大学生物化学工程学院生物医药系

通讯机构：[3]Key Laboratory for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Center for Synthetic & Systems Biology, Institute of Biochemical Engineering, Tsinghua University, Beijing, 100084, China;[1]Beijing Key Laboratory of Biomass Waste Resource Utilization, College of Biochemical Engineering, Beijing Union University, Beijing, 100023, China|[11417144]北京联合大学生物化学工程学院生物医药系;[11417]北京联合大学;[1141726]生物化学工程学院;

年份：2024

卷号：40

期号：9

起止页码：3201-3215

外文期刊名：Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology

收录：Scopus(收录号：2-s2.0-85204940949)

语种：英文

外文关键词：atmospheric and room temperature plasma; high-throughput screening; L-tryptophan; pseudo-fluorescent protein sensor

摘要：L-tryptophan is an indispensable essential amino acid with a wide range of applications, which leads to a high demand. Accordingly, the production of L-tryptophan becomes a much-anticipated direction in research and industrial development. While irrational mutagenesis is an effective means to breed industrial strains, how to screen the strains with desirable phenotypes is still a major challenge. In order to improve the efficiency and accuracy of screening L-tryptophan high-yield strains, we used atmospheric and room temperature plasma mutagenesis to construct a random mutant library and then combined it with high-throughput screening in deep-well plates. Using a pseudo-fluorescent protein sensor capable of responding specifically to L-tryptophan, we successfully screened out a strain producing L-tryptophan at a high yield from a random mutagenesis library. The fermentation with the strain in shake flasks produced L-tryptophan at a yield of 1.99 g/L, which was 41.77% higher than that of the starting strain. Finally, the mechanism of high yield of the strain was deciphered by comparative genomics and transcriptomics. The above strategies provide a solid research foundation for further selection and development of high quality L-tryptophan producing strains. ? 2024 Chinese Academy of Sciences. All rights reserved.