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Open AccessArticle

Application of Adaptive Evolution to Improve the Stability of Bacteriophages during Storage

by Kelvin K. Kering 1,2,3,†, Xiaoxu Zhang 1,2,†, Raphael Nyaruaba 1,2,3, Junping Yu 1 and Hongping Wei 1,3,*
1
Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China
2
University of Chinese Academy of Sciences, Beijing 100049, China
3
Sino-Africa Joint Research Center, Nairobi 00200, Kenya
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Viruses 2020, 12(4), 423; https://doi.org/10.3390/v12040423
Received: 21 January 2020 / Revised: 5 April 2020 / Accepted: 6 April 2020 / Published: 9 April 2020
(This article belongs to the Special Issue Phage Ecology)
Phage stability is important for the successful application of bacteriophages as alternative antibacterial agents. Considering that temperature is a critical factor in phage stability, this study aimed to explore the possibility of improving long-term phage stability through adaptive evolution to elevated temperature. Evolution of three wild-type ancestral phages (Myoviridae phage Wc4 and Podoviridae phages CX5 and P-PSG-11) was induced by subjecting the phages to heat treatment at 60 °C for five cycles. The adapted phages showed better stability than the wild-type ancestral phages when subjected to heat treatment at 60 °C for 1 h and after 60 days of storage at 37 °C. However, the adapted phages could not withstand thermal treatment at 70 °C for 1 h. The infectivity and the lytic properties of the phages were not changed by the evolution process. Whole-genome sequencing revealed that single substitutions in the tail tubular proteins were the only changes observed in the genomes of the adapted phages. This study demonstrates that adaptive evolution could be used as a general method for enhancing the thermal stability of phages without affecting their lytic activity. Sequencing results showed that bacteriophages may exist as a population with minor heterogeneous mutants, which might be important to understand the ecology of phages in different environments. View Full-Text
Keywords: bacteriophage; phage therapy; biocontrol; adaptation; natural selection; thermal stability; genetic variation bacteriophage; phage therapy; biocontrol; adaptation; natural selection; thermal stability; genetic variation
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MDPI and ACS Style

Kering, K.K.; Zhang, X.; Nyaruaba, R.; Yu, J.; Wei, H. Application of Adaptive Evolution to Improve the Stability of Bacteriophages during Storage. Viruses 2020, 12, 423.

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