A Novel Bacteriophage Targeting Cronobacter sakazakii Is a Potential Biocontrol Agent in Foods.
Cronobacter sakazakii is an important pathogen that causes high mortality in infants. Due to its occasional antibiotic resistance, a bacteriophage approach might be an alternative effective method for the control of this pathogen. To develop a novel biocontrol agent using bacteriophages, the C. sakazakii-infecting phage CR5 was newly isolated and characterized. Interestingly, this phage exhibited efficient and relatively durable host lysis activity. In addition, a specific gene knockout study and subsequent complementation experiment revealed that this phage infected the host strain using the bacterial flagella. The complete genome sequence analysis of phage CR5 showed that its genome contains 223,989 bp of DNA, including 231 predicted open reading frames (ORFs), and it has a G+C content of 50.06%. The annotated ORFs were classified into six functional groups (structure, packaging, host lysis, DNA manipulation, transcription, and additional functions); no gene was found to be related to virulence or toxin or lysogen formation, but >80% of the predicted ORFs are unknown. In addition, a phage proteomic analysis using SDS-PAGE and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) revealed that seven phage structural proteins are indeed present, supporting the ORF predictions. To verify the potential of this phage as a biocontrol agent against C. sakazakii, it was added to infant formula milk contaminated with a C. sakazakii clinical isolate or food isolate, revealing complete growth inhibition of the isolates by the addition of phage CR5 when the multiplicity of infection (MOI) was 10(5).
Department of Food Science and Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin, South Korea.
Ju-Hoon
JH
Lee
Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, and Center for Food and Bioconvergence, Seoul National University, Seoul, South Korea.
Jaewoo
J
Bai
Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, and Center for Food and Bioconvergence, Seoul National University, Seoul, South Korea.
Hakdong
H
Shin
Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, and Center for Food and Bioconvergence, Seoul National University, Seoul, South Korea.
Yeran
Y
Kim
Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, and Center for Food and Bioconvergence, Seoul National University, Seoul, South Korea.
Bookyung
B
Park
National Institute of Crop Science, Crop Cultivation and Environment Research Division, RDA, Suwon, South Korea.
Sunggi
S
Heu
Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, and Center for Food and Bioconvergence, Seoul National University, Seoul, South Korea Institute of Food Industrialization, Institute of Green Bio Science & Technology, Seoul National University, Pyeongchang, Gangwon, South Korea sangryu@snu.ac.kr.
Sangryeol
S
Ryu
D001433:Bacteriolysis; D001482:Base Composition; D061046:Biological Control Agents / Q000302:isolation & purification* / Q000378:metabolism; D044083:Cronobacter sakazakii / Q000821:virology*; D004279:DNA, Viral / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005516:Food Microbiology; D016679:Genome, Viral; D006801:Humans; D007223:Infant; D041943:Infant Formula / Q000382:microbiology; D008969:Molecular Sequence Data; D017900:Myoviridae / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology / Q000648:ultrastructure; D016366:Open Reading Frames; D040901:Proteomics; D017422:Sequence Analysis, DNA
D061046:Biological Control Agents; D004279:DNA, Viral