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High-Throughput System for Screening of Cephalosporin C High-Yield Strain by 48-Deep-Well Microtiter Plates

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Abstract

Improvement of microbial strains for the high-production of industrial products has been the hallmark of all commercial fermentation processes. Strain improvement has been conventionally achieved through mutation and selection. However, most of the screenings were performed in shake flasks, which made the screening procedure very complex, time-consuming, and inefficient. Most mutant spore suspension had no chance to be screened due to the low-throughput of shake flasks and had to be sacrificed. In this paper, in order to get a Cephalosporin C (CPC) high-yield stain, traditional mutagenesis was employed to obtain the mutant library and gave them the equal screening chance by a novel mixture culture method combined with high-throughput screening method. The good correlation of fermentation results between differing-scale cultivations confirmed the feasibility of utilizing the 48-deep microtiter plates as a scale-down tool instead of shake flasks for culturing high-aerobic microbes with long cultivation period. The microbioassay based on the antibacterial activity of CPC against Alcaligenes faecalis was used to select mutants. As a result, the high-yield strain W-6 was successfully screened out and the CPC titer was nearly 50 % higher than that of the parental strain in the shake flask. The CPC production of strain W-6 was further validated in 50 l bioreactor, and the CPC production reached 32.0 g/l, twofold higher than that of the wild strain.

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Acknowledgements

This work was financially supported by a grant from the Major State Basic Research Development Program of China (973 Program), No. 2012CB721006, the National High Technology Research and Development Program of China (863 Program), No. 2006AA020302, National Major Scientific Technological Special Project No. 2012YQ 15008709, and National High Technology Research and Development Program of China (863 Program), No.2012AA021201

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, People’s Republic of China

    Jun Tan, Ju Chu, Yuyou Hao, Yuanxin Guo, Yingping Zhuang & Siliang Zhang

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  1. Jun Tan
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  2. Ju Chu
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  3. Yuyou Hao
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  4. Yuanxin Guo
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Correspondence to Ju Chu or Yuyou Hao.

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Tan, J., Chu, J., Hao, Y. et al. High-Throughput System for Screening of Cephalosporin C High-Yield Strain by 48-Deep-Well Microtiter Plates. Appl Biochem Biotechnol 169, 1683–1695 (2013). https://doi.org/10.1007/s12010-013-0095-4

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  • DOI: https://doi.org/10.1007/s12010-013-0095-4

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