Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Korean Society for Microbiology and Biotechnology 2023
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
_version_ 1785070338918318080
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
work_keys_str_mv AT yaozhiming ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT fanjingyan ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT daijun ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT yuchen ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT zenghan ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT liqingzhi ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT huwei ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT yanchaoyue ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT haomeilin ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT lihaotian ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT lishuo ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT liujie ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT huangqi ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT lilu ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT zhourui ahighthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT yaozhiming highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT fanjingyan highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT daijun highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT yuchen highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT zenghan highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT liqingzhi highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT huwei highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT yanchaoyue highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT haomeilin highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT lihaotian highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT lishuo highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT liujie highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT huangqi highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT lilu highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae
AT zhourui highthroughputmethodbasedonmicroculturetechnologyforscreeningofhighyieldstrainsoftylosinproducingstreptomycesfradiae