Culture Collection of Industrial Microorganisms-Microbiological Resources Center, Department of Microbiology, Prof. Wacław Dąbrowski Institute of Agricultural and Food Biotechnology-State Research Institute, Rakowiecka 36 Str., 02-532 Warsaw, Poland.
Laboratory of Biotechnology and Molecular Engineering, Department of Microbiology, Prof. Wacław Dąbrowski Institute of Agricultural and Food Biotechnology-State Research Institute, Rakowiecka 36 Str., 02-532 Warsaw, Poland.
Genes (Basel). 2023 Jun 20;14(6):1303. 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.
该菌产生的愈创木酚和卤代酚等化合物会产生不良风味和气味，从而降低果汁的质量。由于其能够抵抗高温和活性酸度等环境因素，因此，该菌的失活构成了一个挑战。然而，噬菌体的使用似乎是一种很有前途的方法。在这项研究中，我们旨在分离并全面表征一种针对该菌的新型噬菌体。噬菌体菌株 KKP 3916 是从果园土壤中分离出来的，可对抗 KKP 3133 菌株。使用 Bioscreen C Pro 生长分析仪确定细菌宿主的范围以及在不同感染复数 (MOI) 率下添加噬菌体对宿主生长动力学的影响。噬菌体菌株 KKP 3916 在很宽的温度范围（4°C 至 30°C）和活性酸度值（pH 值为 3 至 11）下保持其活性。在 70°C 时，噬菌体的活性降低了 99.9%。相反，在 80°C 时，观察到对细菌宿主没有活性。暴露在紫外线下 30 分钟会使噬菌体的活性降低近 99.99%。基于透射电子显微镜 (TEM) 和全基因组测序 (WGS) 分析，噬菌体菌株 KKP 3916 被归类为有尾噬菌体。基因组测序表明，新分离的噬菌体具有线性双链 DNA（dsDNA），大小为 120 bp 和 131 bp，G+C 含量为 40.3%。在预测的 204 个蛋白中，有 134 个功能未知，其余的被注释为结构、复制和裂解蛋白。在新分离噬菌体的基因组中未发现与抗生素抗性相关的基因。但是，鉴定了几个区域，包括四个与整合到细菌宿主基因组和切除酶相关的区域，这表明该噬菌体具有温和（溶原）的生命周期。由于其潜在参与水平基因转移的风险，该噬菌体不适合进一步研究其在食品生物防治中的应用。据我们所知，这是第一篇关于该菌特异性噬菌体的分离和全基因组分析的文章。
