Characterization and Genome Study of a Newly Isolated Temperate Phage Belonging to a New Genus Targeting Alicyclobacillus acidoterrestris

• Published in: Genes 2023-06, Vol.14 (6), p.1303
• Main Authors: Shymialevich, Dziyana, Wójcicki, Michał, Świder, Olga, Średnicka, Paulina, Sokołowska, Barbara
• Format: Article
• Language: English
• Subjects: Acidic soils; Acidity; Alicyclobacillus; Alicyclobacillus - genetics; Alicyclobacillus acidoterrestris; Antibiotic resistance; Bacteria; Bacteriophages - genetics; Biological control; Carbohydrates; Cellulose acetate; Electron microscopy; Environmental factors; Food; Fruit juices; Gene transfer; Guaiacol; Heat resistance; High temperature; Horizontal transfer; Hot Temperature; Life cycles; Lysis; Microorganisms; New genera; Phages; Temperature; Vegetables; Whole genome sequencing
• ISSN: 2073-4425; 2073-4425
• DOI: 10.3390/genes14061303

The spoilage of juices by spp. remains a serious problem in industry and leads to economic losses. Compounds such as guaiacol and halophenols, which are produced by , create undesirable flavors and odors and, thus, decrease the quality of juices. The inactivation of spp. constitutes a challenge because it is resistant to environmental factors, such as high temperatures, and active acidity. However, the use of bacteriophages seems to be a promising approach. In this study, we aimed to isolate and comprehensively characterize a novel bacteriophage targeting spp. The phage strain KKP 3916 was isolated from orchard soil against the strain KKP 3133. The bacterial host's range and the effect of phage addition at different rates of multiplicity of infections (MOIs) on the host's growth kinetics were determined using a Bioscreen C Pro growth analyzer. The phage strain KKP 3916, retained its activity in a wide range of temperatures (from 4 °C to 30 °C) and active acidity values (pH from 3 to 11). At 70 °C, the activity of the phage decreased by 99.9%. In turn, at 80 °C, no activity against the bacterial host was observed. Thirty minutes of exposure to UV reduced the activity of the phages by almost 99.99%. Based on transmission-electron microscopy (TEM) and whole-genome sequencing (WGS) analyses, the phage strain KKP 3916 was classified as a tailed bacteriophage. The genomic sequencing revealed that the newly isolated phage had linear double-stranded DNA (dsDNA) with sizes of 120 bp and 131 bp and 40.3% G+C content. Of the 204 predicted proteins, 134 were of unknown function, while the remainder were annotated as structural, replication, and lysis proteins. No genes associated with antibiotic resistance were found in the genome of the newly isolated phage. However, several regions, including four associated with integration into the bacterial host genome and excisionase, were identified, which indicates the temperate (lysogenic) life cycle of the bacteriophage. Due to the risk of its potential involvement in horizontal gene transfer, this phage is not an appropriate candidate for further research on its use in food biocontrol. To the best of our knowledge, this is the first article on the isolation and whole-genome analysis of the -specific phage.