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Genome Shuffling and Gentamicin-Resistance to Improve ε-Poly-l-Lysine Productivity of Streptomyces albulus W-156

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Abstract

Genome shuffling has been a recently effective method for screening the desirable phenotypes of industrial strains. Here, we combined genome shuffling and gentamicin resistance to improve the production of ε-poly-l-lysine in Streptomyces albulus W-156. Five starting mutants with higher ε-poly-l-lysine (ε-PL) productivities were firstly obtained by atmospheric and room temperature plasma (ARTP) mutagenesis. After three rounds of genome shuffling with increasing concentration of gentamicin for selection, S. albulus AG3-28, was finally got with a production of 3.43 g/L in shaking flask. In a 5-L fermenter, AG3-28 exhibited a higher ε-PL productivity (56.5 g/L) than the initial strain W-156 (37.5 g/L). Key enzyme activities in primary and secondary metabolic pathways were analyzed, and the transcription levels of hrdD and pls were determined by quantitative real time-polymerase chain reaction (qRT-PCR). Increase of key enzyme activities and the upregulation of the gene transcriptional levels demonstrated that ε-PL synthetic pathway in AG3-28 was obviously strengthened, which might be responsible for the high productivity. Moreover, hyper-yield strain AG3-28 was found to produce a slightly lower ε-PL polymerization degree than the parent strain. Amplified fragment length polymorphism (AFLP) analysis reflects the genetic diversity among the derivates after genome shuffling.

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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 (21376106), the Innovation Plan of Jiangsu Province (KYLX15_1146), the Fundamental Research Funds for the Central Universities (JUSRP51504), the Open Project Program of the Key Laboratory of Industrial Biotechnology, Ministry of Education, China (KLIBKF201302), and the Jiangsu Province Collaborative Innovation Center for Advanced Industrial Fermentation Industry Development Program.

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Authors and Affiliations

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

    Liang Wang, Xusheng Chen, Guangyao Wu, Xin Zeng, Xidong Ren, Lei Tang & Zhonggui Mao

  2. Xiwang Group Company Limited, Zouping, 256209, China

    Shu Li

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  1. Liang Wang
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  2. Xusheng Chen
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  3. Guangyao Wu
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  4. Xin Zeng
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Correspondence to Xusheng Chen or Zhonggui Mao.

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Wang, L., Chen, X., Wu, G. et al. Genome Shuffling and Gentamicin-Resistance to Improve ε-Poly-l-Lysine Productivity of Streptomyces albulus W-156. Appl Biochem Biotechnol 180, 1601–1617 (2016). https://doi.org/10.1007/s12010-016-2190-9

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  • DOI: https://doi.org/10.1007/s12010-016-2190-9

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