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A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae
Tylosin is a potent veterinary macrolide antibiotic produced by the fermentation of Streptomyces fradiae; however, it is necessary to modify S. fradiae strains to improve tylosin production. In this study, we established a high-throughput, 24-well plate screening method for identifying S. fradiae st...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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The Korean Society for Microbiology and Biotechnology
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10331940/ https://www.ncbi.nlm.nih.gov/pubmed/36994618 http://dx.doi.org/10.4014/jmb.2210.10023 |
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author | Yao, Zhiming Fan, Jingyan Dai, Jun Yu, Chen Zeng, Han Li, Qingzhi Hu, Wei Yan, Chaoyue Hao, Meilin Li, Haotian Li, Shuo Liu, Jie Huang, Qi Li, Lu Zhou, Rui |
author_facet | Yao, Zhiming Fan, Jingyan Dai, Jun Yu, Chen Zeng, Han Li, Qingzhi Hu, Wei Yan, Chaoyue Hao, Meilin Li, Haotian Li, Shuo Liu, Jie Huang, Qi Li, Lu Zhou, Rui |
author_sort | Yao, Zhiming |
collection | PubMed |
description | Tylosin is a potent veterinary macrolide antibiotic produced by the fermentation of Streptomyces fradiae; however, it is necessary to modify S. fradiae strains to improve tylosin production. In this study, we established a high-throughput, 24-well plate screening method for identifying S. fradiae strains that produce increased yields of tylosin. Additionally, we constructed mutant libraries of S. fradiae via ultraviolet (UV) irradiation and/or sodium nitrite mutagenesis. A primary screening of the libraries in 24-well plates and UV spectrophotometry identified S. fradiae mutants producing increased yields of tylosin. Mutants with tylosin yield 10% higher than the wild-type strain were inoculated into shake flasks, and the tylosin concentrations produced were determined by high-performance liquid chromatography (HPLC). Joint (UV irradiation and sodium nitrite) mutagenesis resulted in higher yields of mutants with enhanced tylosin production. Finally, 10 mutants showing higher tylosin yield were re-screened in shake flasks. The yield of tylosin A by strains UN-C183 (6767.64 ± 82.43 μg/ml) and UN-C137 (6889.72 ± 70.25 μg/ml) was significantly higher than that of the wild-type strain (6617.99 ± 22.67 μg/ml). These mutant strains will form the basis for further strain breeding in tylosin production. |
format | Online Article Text |
id | pubmed-10331940 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | The Korean Society for Microbiology and Biotechnology |
record_format | MEDLINE/PubMed |
spelling | pubmed-103319402023-07-11 A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae Yao, Zhiming Fan, Jingyan Dai, Jun Yu, Chen Zeng, Han Li, Qingzhi Hu, Wei Yan, Chaoyue Hao, Meilin Li, Haotian Li, Shuo Liu, Jie Huang, Qi Li, Lu Zhou, Rui J Microbiol Biotechnol Research article Tylosin is a potent veterinary macrolide antibiotic produced by the fermentation of Streptomyces fradiae; however, it is necessary to modify S. fradiae strains to improve tylosin production. In this study, we established a high-throughput, 24-well plate screening method for identifying S. fradiae strains that produce increased yields of tylosin. Additionally, we constructed mutant libraries of S. fradiae via ultraviolet (UV) irradiation and/or sodium nitrite mutagenesis. A primary screening of the libraries in 24-well plates and UV spectrophotometry identified S. fradiae mutants producing increased yields of tylosin. Mutants with tylosin yield 10% higher than the wild-type strain were inoculated into shake flasks, and the tylosin concentrations produced were determined by high-performance liquid chromatography (HPLC). Joint (UV irradiation and sodium nitrite) mutagenesis resulted in higher yields of mutants with enhanced tylosin production. Finally, 10 mutants showing higher tylosin yield were re-screened in shake flasks. The yield of tylosin A by strains UN-C183 (6767.64 ± 82.43 μg/ml) and UN-C137 (6889.72 ± 70.25 μg/ml) was significantly higher than that of the wild-type strain (6617.99 ± 22.67 μg/ml). These mutant strains will form the basis for further strain breeding in tylosin production. The Korean Society for Microbiology and Biotechnology 2023-06-28 2023-03-10 /pmc/articles/PMC10331940/ /pubmed/36994618 http://dx.doi.org/10.4014/jmb.2210.10023 Text en Copyright © 2023 by the authors. Licensee KMB https://creativecommons.org/licenses/by/4.0/This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/) |
spellingShingle | Research article Yao, Zhiming Fan, Jingyan Dai, Jun Yu, Chen Zeng, Han Li, Qingzhi Hu, Wei Yan, Chaoyue Hao, Meilin Li, Haotian Li, Shuo Liu, Jie Huang, Qi Li, Lu Zhou, Rui A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae |
title | A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae |
title_full | A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae |
title_fullStr | A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae |
title_full_unstemmed | A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae |
title_short | A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae |
title_sort | high-throughput method based on microculture technology for screening of high-yield strains of tylosin-producing streptomyces fradiae |
topic | Research article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10331940/ https://www.ncbi.nlm.nih.gov/pubmed/36994618 http://dx.doi.org/10.4014/jmb.2210.10023 |
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