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Shin, Hakdong,Lee, Ju-Hoon,Yoon, Hyunjin,Kang, Dong-Hyun,Ryu, Sangryeol American Society for Microbiology 2014 Applied and environmental microbiology Vol.80 No.1
<P>To understand phage infection and host cell lysis mechanisms in pathogenic <I>Salmonella</I>, a novel <I>Salmonella enterica</I> serovar Typhimurium-targeting bacteriophage, SPN9CC, belonging to the <I>Podoviridae</I> family was isolated and characterized. The phage infects <I>S</I>. Typhimurium via the O antigen of lipopolysaccharide (LPS) and forms clear plaques with cloudy centers due to lysogen formation. Phylogenetic analysis of phage major capsid proteins revealed that this phage is a member of the lysogen-forming P22-like phage group. However, comparative genomic analysis of SPN9CC with P22-like phages indicated that their lysogeny control regions and host cell lysis gene clusters show very low levels of identity, suggesting that lysogen formation and host cell lysis mechanisms may be diverse among phages in this group. Analysis of the expression of SPN9CC host cell lysis genes encoding holin, endolysin, and Rz/Rz1-like proteins individually or in combinations in <I>S</I>. Typhimurium and <I>Escherichia coli</I> hosts revealed that collaboration of these lysis proteins is important for the lysis of both hosts and that holin is a key protein. To further investigate the role of the lysogeny control region in phage SPN9CC, a Δ<I>cI</I> mutant (SPN9CCM) of phage SPN9CC was constructed. The mutant does not produce a cloudy center in the plaques, suggesting that this mutant phage is virulent and no longer temperate. Subsequent comparative one-step growth analysis and challenge assays revealed that SPN9CCM has shorter eclipse/latency periods and a larger burst size, as well as higher host cell lysis activity, than SPN9CC. The present work indicates the possibility of engineering temperate phages as promising biocontrol agents similar to virulent phages.</P>
Choi, Younho,Shin, Hakdong,Lee, Ju-Hoon,Ryu, Sangryeol American Society for Microbiology 2013 Applied and environmental microbiology Vol.79 No.16
<P>A novel flagellatropic phage of <I>Salmonella enterica</I> serovar Typhimurium, called iEPS5, was isolated and characterized. iEPS5 has an icosahedral head and a long noncontractile tail with a tail fiber. Genome sequencing revealed a double-stranded DNA of 59,254 bp having 73 open reading frames (ORFs). To identify the receptor for iEPS5, Tn<I>5</I> transposon insertion mutants of <I>S</I>. Typhimurium SL1344 that were resistant to the phage were isolated. All of the phage-resistant mutants were found to have mutations in genes involved in flagellar formation, suggesting that the flagellum is the adsorption target of this phage. Analysis of phage infection using the Δ<I>motA</I> mutant, which is flagellated but nonmotile, demonstrated the requirement of flagellar rotation for iEPS5 infection. Further analysis of phage infection using the Δ<I>cheY</I> mutant revealed that iEPS5 could infect host bacteria only when the flagellum is rotating counterclockwise (CCW). These results suggested that the CCW-rotating flagellar filament is essential for phage adsorption and required for successful infection by iEPS5. In contrast to the well-studied flagellatropic phage Chi, iEPS5 cannot infect the Δ<I>fliK</I> mutant that makes a polyhook without a flagellar filament, suggesting that these two flagellatropic phages utilize different infection mechanisms. Here, we present evidence that iEPS5 injects its DNA into the flagellar filament for infection by assessing DNA transfer from SYBR gold-labeled iEPS5 to the host bacteria.</P>
Lee, Ju-Hoon,Shin, Hakdong,Ji, Samnyu,Malhotra, Shweta,Kumar, Mukesh,Ryu, Sangryeol,Heu, Sunggi American Society for Microbiology 2012 Journal of virology Vol.86 No.16
<P>Pectobacterium carotovorumsubsp.carotovorumis a phytopathogen causing soft rot disease on diverse plant species. To control this plant pathogen,P. carotovorumsubsp.carotovorum-targeting bacteriophage PP1 was isolated and its genome was completely sequenced to develop a novel biocontrol agent. Interestingly, the 44,400-bp genome sequence does not encode any gene involved in the formation of lysogen, suggesting that this phage may be very useful as a biocontrol agent because it does not make lysogen after host infection. This is the first report on the complete genome sequence of theP. carotovorumsubsp.carotovorum-targeting bacteriophage, and it will enhance our understanding of the interaction between phytopathogens and their targeting bacteriophages.</P>
<i>Vibrio vulnificus</i> Bacteriophage SSP002 as a Possible Biocontrol Agent
Lee, Hyun Sung,Choi, Slae,Shin, Hakdong,Lee, Ju-Hoon,Choi, Sang Ho American Society for Microbiology 2014 Applied and environmental microbiology Vol.80 No.2
<P>A novel <I>Vibrio vulnificus</I>-infecting bacteriophage, SSP002, belonging to the <I>Siphoviridae</I> family, was isolated from the coastal area of the Yellow Sea of South Korea. Host range analysis revealed that the growth inhibition of phage SSP002 is relatively specific to <I>V. vulnificus</I> strains from both clinical and environmental samples. In addition, a one-step growth curve analysis and a bacteriophage stability test revealed a latent period of 65 min, a burst size of 23 ± 2 PFU, as well as broad temperature (20°C to 60°C) and pH stability (pH 3 to 12) ranges. A Tn<I>5</I> random transposon mutation of <I>V. vulnificus</I> and partial DNA sequencing of the inserted Tn<I>5</I> regions revealed that the <I>flhA</I>, <I>flhB</I>, <I>fliF</I>, and <I>fleQ</I> mutants are resistant to SSP002 phage infection, suggesting that the flagellum may be the host receptor for infection. The subsequent construction of specific gene-inactivated mutants (<I>flhA</I>, <I>flhB</I>, <I>fliF</I>, and <I>fleQ</I>) and complementation experiments substantiated this. Previously, the genome of phage SSP002 was completely sequenced and analyzed. Comparative genomic analysis of phage SSP002 and <I>Vibrio parahaemolyticus</I> phage vB_VpaS_MAR10 showed differences among their tail-related genes, supporting different host ranges at the species level, even though their genome sequences are highly similar. An additional mouse survival test showed that the administration of phage SSP002 at a multiplicity of infection of 1,000 significantly protects mice from infection by <I>V. vulnificus</I> for up to 2 months, suggesting that this phage may be a good candidate for the development of biocontrol agents against <I>V. vulnificus</I> infection.</P>
Evaluation of Prebiotic Potential of Soy Protein Isolate-Based Imitation Cheeses
Minju Song,Eunji Cho,Gyungcheon Kim,Hakdong Shin,Nambae Jeon,Honghoon Yoo,Young-Wan Kim 한국식품영양과학회 2021 한국식품영양과학회 학술대회발표집 Vol.2021 No.10
As alternative dairy products, soy protein isolate-based imitation cheese had been developed by addition of hydrocolloids to modulate the texture. In this study, it was attempted to introduce healthful property of the soy cheese by the addition of prebiotics. A soy cheese containing of either one of prebiotics (kestose, inulin, fructooligosaccharides) or sucrose (as a control) was prepared. After in vitro digestion, the samples were mixed with basal medium, followed by anaerobic fermentation. The changes in gut microbiota were monitored by 16S rRNA analysis using NGS. The microbial diversity was decreased in samples with soy cheese, and the species composition across space was significantly changed compared to controls. The populations of health promoting bacteria increased, whereas the pathogenic bacteria decreased by supplementation of the soy cheeses. Interestingly, there was no significant difference between sucrose-added and prebiotics-added soy cheeses. In conclusion, the soy cheese with hydrocolloids not only exhibits textural properties similar to the commercial cheeses, but also would have health-promoting potentials due to the growth induction for probiotic bacteria.
( Sooyeon Lim ),( Dong Hoon Lee ),( Woori Kwak ),( Hakdong Shin ),( Hye Jin Ku ),( Jong Eun Lee ),( Gun Eui Lee ),( Heebal Kim ),( Sang Ho Choi ),( Sangryeol Ryu ),( Ju Hoon Lee ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.1
Staphylococcus aureus is an important foodborne pathogen that causes diverse diseases ranging from minor infections to life-threatening conditions in humans and animals. To further understand its pathogenesis, the genome of the strain S. aureus FORC_001 was isolated from a contaminated food. Its genome consists of 2,886,017 bp double-stranded DNA with a GC content of 32.8%. It is predicted to contain 2,728 open reading frames, 57 tRNAs, and 6 rRNA operons, including 1 additional 5S rRNA gene. Comparative phylogenetic tree analysis of 40 complete S. aureus genome sequences using average nucleotide identity (ANI) revealed that strain FORC_001 belonged to Group I. The closest phylogenetic match was S. aureus MRSA252, according to a whole-genome ANI (99.87%), suggesting that they might share a common ancestor. Comparative genome analysis of FORC_001 and MRSA252 revealed two non-homologous regions: Regions I and II. The presence of various antibiotic resistance genes, including the SCCmec cluster in Region I of MRSA252, suggests that this strain might have acquired the SCCmec cluster to adapt to specific environments containing methicillin. Region II of both genomes contains prophage regions but their DNA sequence identity is very low, suggesting that the prophages might differ. This is the first report of the complete genome sequence of S. aureus isolated from a real foodborne outbreak in South Korea. This report would be helpful to extend our understanding about the genome, general characteristics, and virulence factors of S. aureus for further studies of pathogenesis, rapid detection, and epidemiological investigation in foodborne outbreak.
( Hyeri Kim ),( Jae Hyoung Cho ),( Jin Ho Cho ),( Minho Song ),( Hakdong Shin ),( Sheena Kim ),( Eun Sol Kim ),( Hyeun Bum Kim ),( Ju-hoon Lee ) 한국축산학회(구 한국동물자원과학회) 2021 한국축산학회지 Vol.63 No.1
Salmonella enterica is a representative foodborne pathogen in the world. The S. enterica strain K_SA184 was isolated from the lamb (Ovis aries), which was collected from a local traditional market in South Korea. In this study, the S. enterica strain K_SA184 was sequenced using PacBio RS II and Illumina NextSeq 500 platforms. The final complete genome of the S. enterica strain K_SA184 consist of one circular chromosome (4,725,087 bp) with 52.3% of guanine + cytosine (G + C) content, 4,363 of coding sequence (CDS), 85 of tRNA, and 22 of rRNA genes. The S. enterica strain K_SA184 genome includes encoding virulence genes, such as Type III secretion systems and multidrug resistance related genes.