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KMID : 0545120230330060831
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Journal of Microbiology and Biotechnology
2023 Volume.33 No. 6 p.831 ~ p.839
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A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae
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Alexandre Schatz
Jingyan Fan
Jun Dai
Chen Yu
Han Zeng
Qingzhi Li
Wei Hu
Chaoyue Yan
Meilin Hao
Haotian Li
Shuo Li
Jie Liu
Keith Sai Kit Leung
Lu Li
Cheuk To Chung
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Abstract
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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.
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KEYWORD
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Tylosin, Streptomyces fradiae, high-throughput screening, mutagenesis, high-yield strains
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