Abstract
The Escherichia coli (E. coli) has been widely employed in biotechnology industry and academia. However, the bioproduct manufacturing driven by E. coli is prone to the phage contamination. Good laboratory/factory hygiene may decrease but not avoid completely the chances of the phage contamination. The present study aims to resolve this problem by engineering laboratory/factory-specific phage-resistant E. coli strains. By adding a laboratory or factory derived phage into the atmospheric and room temperature plasma mutagenized E. coli, a phage-resistant strain could be generated. Interestingly, the resistant strain exhibited cross-resistance to unencountered phages. When operating the resistant strain in a polluted environment, the phage contamination was largely prevented. There was no significant difference in heterogeneous protein production between the parental strain and the phage-resistant strain. Importantly, it requires only one day to generate the phage-resistant strain. This practical method for engineering laboratory/factory-specific phage-resistant strains may have great potential in resuming E. coli operation in laboratories and factories during phage contamination outbreaks.
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Data availability
The codon-optimized Haemophilus ducreyi nicotinamide phosphoribosyltransferase with a C-terminal six histidine tag was deposited at GenBank (Accession No: MW759281). The sequencing files in fastq format obtained from whole-genome sequencing of the strain Hd1 were submitted to the NCBI under SRA accession number PRJNA783137. The GenBank accession numbers for the E. coli BL21 reference genome were CP060121.1, CP053602.1, and AM946981.2.
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Acknowledgements
This research was supported by the National Key Research and Development Program of China (2020YFC210007), the Fundamental Research Funds for the Provincial Universities of Zhejiang (RF-C2020002), and the Major Research program of Zhejiang Provincial National Natural Science Foundation of China (LD21C050001).
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This research was supported by the National Key Research and Development Program of China (2020YFC210007), the Fundamental Research Funds for the Provincial Universities of Zhejiang (RF-C2020002), and the Major Research program of Zhejiang Provincial National Natural Science Foundation of China (LD21C050001).
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YPX, YGZ, and QS conceived and supervised the project. QS, XTZ, and QG carried out all the experiments and data analyses. QS and YPX are responsible for the preparation and revision of the manuscript. All authors read and approved the final manuscript.
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Shen, Q., Zhou, XT., Guo, Q. et al. Engineering laboratory/factory-specific phage-resistant strains of Escherichia coli by mutagenesis and screening. World J Microbiol Biotechnol 38, 51 (2022). https://doi.org/10.1007/s11274-022-03239-y
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DOI: https://doi.org/10.1007/s11274-022-03239-y