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Development of Microtiter Plate Culture Method for Rapid Screening of ε-Poly-L-Lysine-Producing Strains

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Abstract

ε-Poly-l-lysine (ε-PL) produced by Streptomyces albulus possesses a broad spectrum of antimicrobial activity and is widely used as a food preservative. To extensively screen ε-PL-overproducing strain, we developed an integrated high-throughput screening assay using ribosome engineering technology. The production protocol was scaled down to 24- and 48-deep-well microtiter plates (MTPs). The microplate reader assay was used to monitor ε-PL production. A good correlation was observed between the fermentation results obtained in both 24-(48)-deep-well MTPs and conventional Erlenmeyer flasks. Using this protocol, the production of ε-PL in an entire MTP was determined in <5 min without compromising on accuracy. The high-yielding strain selected through this protocol was also tested in Erlenmeyer flasks. The result showed that the ε-PL production of the high-yielding mutants was nearly 45% higher than that of the parent stain. Thus, development of this protocol is expected to accelerate the selection of ε-PL-overproducing strains.

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Acknowledgments

This work was supported by the Cooperation Project of Jiangsu Province among Industries, Universities and Institutes (BY2016022-25), the Program of the National Natural Science Foundation of China (31671846,31301556), and the Jiangsu Province Collaborative Innovation Center for Advanced Industrial Fermentation Industry Development Program.

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  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China

    Yong-Juan Liu, Xu-Sheng Chen, Jun-Jie Zhao, Long Pan & Zhong-Gui Mao

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  1. Yong-Juan Liu
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  2. Xu-Sheng Chen
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  3. Jun-Jie Zhao
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  4. Long Pan
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Correspondence to Xu-Sheng Chen or Zhong-Gui Mao.

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Liu, YJ., Chen, XS., Zhao, JJ. et al. Development of Microtiter Plate Culture Method for Rapid Screening of ε-Poly-L-Lysine-Producing Strains. Appl Biochem Biotechnol 183, 1209–1223 (2017). https://doi.org/10.1007/s12010-017-2493-5

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