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Genome Shuffling of Penicillium citrinum for Enhanced Production of Nuclease P1

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Abstract

Genome shuffling is a powerful approach for efficiently engineering industrial microbial strains with interested phenotypes. Here we reported a high producer of nuclease P1, Penicillium citrinum G-16, that was bred by the classical physics-mutagenesis and genome shuffling process. The starting populations were generated by 60Co γ-irradiation mutagenesis. The derived two protoplast fractions were inactivated by heat-treatment and ultraviolet radiation respectively, then mixed together and subjected to recursive protoplast fusion. Three recombinants, E-16, F-71, and G-16, were roughly obtained from six cycles of genome shuffling. The activity of nuclease P1 by recombinant G-16 was improved up to 1,980.22 U4/ml in a 5-l fermentor, which was 4.7-fold higher than that of the starting strain. The sporulation of recombinant G-16 was distinguished from the starting strain. Random amplified polymorphic DNA assay revealed genotypic differences between the shuffled strains and the wild type strain. The close similarity among the high producers suggested that the genetic basis of high-yield strains was achieved by genome shuffling.

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Acknowledgments

The work was financially supported by the National Nature and Science Foundation of China (3117175) and Nature and Science Foundation of Zhejiang Province (Y3100609).

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The authors declare that they have no conflict of interests.

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Authors and Affiliations

  1. Department of Biological Engineering, School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, 310025, China

    Chao Wang, Gongzhen Wu, Yudong Li, Yinyin Huang & Xinle Liang

  2. Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA

    Fuming Zhang

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  1. Chao Wang
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  2. Gongzhen Wu
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  6. Xinle Liang
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Correspondence to Xinle Liang.

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Wang, C., Wu, G., Li, Y. et al. Genome Shuffling of Penicillium citrinum for Enhanced Production of Nuclease P1. Appl Biochem Biotechnol 170, 1533–1545 (2013). https://doi.org/10.1007/s12010-013-0297-9

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  • DOI: https://doi.org/10.1007/s12010-013-0297-9

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