Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae

  • Zhiming Yao (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Jingyan Fan (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Jun Dai (Hubei Provincial Bioengineering Technology Research Center for Animal Health Products) ;
  • Chen Yu (Hubei Provincial Bioengineering Technology Research Center for Animal Health Products) ;
  • Han Zeng (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Qingzhi Li (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Wei Hu (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Chaoyue Yan (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Meilin Hao (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Haotian Li (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Shuo Li (Hubei Provincial Bioengineering Technology Research Center for Animal Health Products) ;
  • Jie Liu (Hubei Provincial Bioengineering Technology Research Center for Animal Health Products) ;
  • Qi Huang (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Lu Li (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University) ;
  • Rui Zhou (National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University)
  • Received : 2022.10.17
  • Accepted : 2023.02.28
  • Published : 2023.06.28
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Abstract

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 ㎍/ml) and UN-C137 (6889.72 ± 70.25 ㎍/ml) was significantly higher than that of the wild-type strain (6617.99 ± 22.67 ㎍/ml). These mutant strains will form the basis for further strain breeding in tylosin production.

Keywords

Acknowledgement

This research was funded by the Hubei Province Technology Innovation Project (2019AEE005). We thank the Hubei Provincial Bioengineering Technology Research Center for Animal Health Products for their technical support and Prof. Paul R. Langford from Imperial College London for language modifications to the manuscript.

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