• Journal of Microbiology
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• v.35 no.4
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• pp.347-353
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• 1997

We have characterized a phage library displaying random 22mer peptides which were produced as N-terminal fusions to the pIII coat protein of M13 filamentous phages. Among the sixty phages randomly picked from the library, 25 phages had the 22mer peptide inserts. The DNA sequence analysis of the 25 inserts showed the following results: first, each nucleotide was represented almost equally at each codon position except that there were some biases toward G bases at the first position of the codons. Secondly, the expected 47 sense codons were represented. The deduced amino acid sequences of the 25 inserts were analyzed to examine its diversity. Glycine and glutamate were the two most overrepresented residues above the expected value, whereas cysteine and threonine residues were underrepresented. The range of dicersity in dipeptide sequences showed that the amino acid residues were randomly distributed along the peptide insert. Acidic, basic, polar, and nonpolar amino acid residues were represented to the extent expected at most positions of the peptide inserts. The predicted isoelectric points and hydropathy indices of the 25 peptides showed that a variety of the peptide were represented in the library. These results indicate that this phage display library could be useful in fiuding ligands for a broad spectrum of receptors by affinity screening.

• Food Science of Animal Resources
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• v.37 no.2
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• pp.320-328
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• 2017

In this study, two Listeria bacteriophages, LMP1 and LMP7, were isolated from chicken feces as a means of biocontrol of L. monocytogenes. Both bacteriophages had a lytic effect on L. monocytogenes ATCC 7644, 15313, 19114, and 19115. Phages LMP1 and LMP7 were able to inhibit the growth of L. monocytogenes ATCC 7644 and 19114 in tryptic soy broth at $10^{\circ}C$ and $30^{\circ}C$. Nevertheless, LMP1 was more effective than LMP7 at inhibiting L. monocytogenes ATCC 19114. On the contrary, LMP7 was more effective than LMP1 at inhibiting L. monocytogenes ATCC 7644. The morphology of LMP1 and LMP7 resembled that of members of the Siphoviridae family. The growth of L. monocytogenes ATCC 7644 was inhibited by both LMP1 and LMP7 in milk; however, the growth of L. monocytogenes ATCC 19114 was only inhibited by LMP1 at $30^{\circ}C$. The lytic activity of bacteriophages was also evaluated at $4^{\circ}C$ in milk in order to investigate the potential use of these phages in refrigerated products. In conclusion, these two bacteriophages exhibit different host specificities and characteristics, suggesting that they can be used as a component of a phage cocktail to control L. monocytogenes in the food industry.

• Korean Journal of Microbiology
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• v.23 no.2
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• pp.115-121
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• 1985

Five representative virulent phages (J1, TK93, K1, PD5, and CP1) and one temperate phage (.phi.1043) of Lactobacillus casei were compared to each other by analyzing the agarose gel electrophoretic patterns of restriction enzyme-digested phage DNAs. Nucleic acids of all the tested phages were double stranded DNA. DNAs of J1, TK93, K1, and ${\phi}$ 1043 phages had a size of about 42kb, but the size of PD5 and CP1 DNAs was avout 140kb. J1, TK93, K1, PD5, CP1, and ${\phi}$ 1043 DNAs were digested to 13, 13, 11, 14, 14, and 12 fragments by EcoR1, respectively, and showed its characteristec restriction patterns. Cohesive ends were present in J1, TK93, and ${\phi}$ 1043, but were absent in K1, PD5, and CP1. Restriction maps of J1 and TK93 DNAs showed nearly complete homology and their evolutionary relationship based upon the restriction analysis was discussed.

• Korean Journal of Food Science and Technology
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• v.25 no.1
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• pp.46-51
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• 1993

The inactivating effect of iron(II)-ascorbate complex (Fe-Asc) on various phages was previously reported. This paper describes the molecular target in the phage virion attacked by Fe-Asc. The effect of Fe-Asc on protein was investigated with bovine serum albumin and the structural protein of phage J1. There were no differences in the SDS-polyacrylamide gel electrophoresis (patterns of these two proteins when either they were treated) with Fe-Asc or not. Also, there were no changes in the amino acid composition and ultraviolet spectrum of the proteins. The effects of Fe-Asc on DNA was investigated with pUC18 DNA, M13mpB DNA and ${\lambda}$ DNA as well as DNA from phage J1. Fe-Asc caused initially nicking of the subsequently form of pUC18 DNA to yield the open circular form and then subsequently the linear form. Strand breaks were also confirmed with M13mp8 DNA and ${\lambda}$ DNA as well as J1 DNA. The results indicate that the strand breaks in phage DNA could be responsible for the inactivation of phages by Fe-Asc.

• Journal of fish pathology
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• v.11 no.2
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• pp.137-141
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• 1998

Temperate phages were effectively induced from presumptive lysogenized cells of 96 strains out of 111 strains of L. garvieae No. 44 strains (phage type B) as the host cell. Similar cultures in distilled water-based TSB did not induce lytic infection in these cells. These temperate phages were also effectively induced by ultraviolet irradiation. All phages isolated were lytic only to L. garvieae No. 44 strain and the lytic nature was different from those of PLgY, PLgW, and PLgS. The virions appeared extracellularly after 1h of induction culture and increased in number until reaching the maximum of $10^6$ PFU/ml after 12h. This phage production was lower than that ($10^{10}$ PFU/ml) of the virulent phage.

• YAKHAK HOEJI
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• v.57 no.2
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• pp.125-131
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• 2013

Biologically active peptides, including growth factors and cytokines, participate in various biological processes in human skin. They could provide a great advantage of maintaining healthy skin. Many peptide growth factors like epidermal growth factor (EGF) and human growth hormone (hGH) have been used in cosmetic formulations. The delivery of peptide growth factors across the Stratum corneum, however, seems not sufficient because of their physical properties such as high molecular weight and hydrophilicity. So increasing the penetration of growth factors of interest into skin would be a major concern for ensuring their maximum biological efficacy. In this study, we have identified several skin penetration-enhancing peptides which facilitate delivery of growth factors, when fused at N-terminus of the target protein, into skin. For efficient and rapid screening, we constructed a skin-penetrating assay system using Franz cell and porcine skin. Next, we carried out phage display screening using M-13 bacteriophage with random 12 -amino acid library on its coat protein P3 on that system. After several selection rounds, peptide sequences facilitate the penetration of phages through the porcine skin were identified from a large population of phages. We found that phages with the most potent peptide (S3-2, NGSLNTHLAPIL) could penetrate the porcine skin eight times more than those with control peptide (12 mino acids scrambled peptide). Furthermore, growth factors conjugated with S3-2 peptide penetrate porcine skin three to five times efficiently than non-conjugated growth factors. In conclusion, our data shows that the skin penetration-enhancing peptide we have characterized could increase the delivery of growth factors and is useful for cosmeceutical application.

• The Plant Pathology Journal
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• v.18 no.6
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• pp.333-337
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• 2002

Bacteriophage of Xanthomonas axonopodis pv. citri, a causal agent of citrus canker disease, was studied for its cultural characteristics. The relative efficiency of plat-ing (EOP) of 11 phages used to 13 strains off, axonopodis pv. citri tested ranged from 0.8 to 1, indicating that the phages are homogeneous. Homogeneity of the phages suggests that citrusphage belongs to a single group CPK as reported in a previous study. Typical one-step growth of a phage P5 selected from the citrusphages was observed. The EOP of the P5 was dependent upon the media, pH, and temperature. It was observed that multiplication of the phage cultured in Wakimotos potato semisynthetic media at $25^{\circ}C$ was more effective than that in other temperatures, regardless of the bacterial strains and media used. It was observed that pH 6.5 is optimal for multiplication of the phage. In comparison of the EOP among citrusphages $CP_1$, $CP_2$, and P5, multiplicative characteristic of phage P5 in the bacteria on time-course was similar with that of phage $CP_1$. Thus, it was concluded that citrusphage group CPK from Korea is $CP_1$ based on host specificity of the phage as described in a previous study, homogeneity, and its multiplication pattern.

• Journal of Applied Biological Chemistry
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• v.62 no.3
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• pp.287-292
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• 2019

Pseudomonas tolaasii causes brown blotch disease on the oyster mushroom (Pleurotus ostreatus). Various pathogenic strains of P. tolaasii were isolated and divided into three subtypes, $P1{\alpha}$, $P1{\beta}$, and $P1{\gamma}$. For phage therapy, bacteriophages against to these subtype strains were applied to mushroom cultivation and very successful to prevent from the disease. In this study, bacteriophages were isolated against the representative strains of subtype pathogens and their polyclonal antibodies were synthesized to investigate structural relationship among capsid proteins of phages. Phage preparations over $10^{10}pfu/mL$ were injected to rabbit thigh muscle and polyclonal antibodies were obtained after three times of boost injection. Titers of the antibodies obtained were over $2{\times}10^7Ab/mL$ for the phage ${\phi}6264$, $1{\times}10^6Ab/mL$ for the phage ${\phi}HK2$, and $1{\times}10^7Ab/mL$ for the phage ${\phi}HK19$ and phage ${\phi}HK23$. High specific activities were observed between antibodies and the corresponding bacteriophages. Some cross-reactivities between the antibodies and non-corresponding bacteriophages were also measured. Antibody $Ab{\phi}6264$ inactivated all phages of $P1{\alpha}$ subtype and only phage ${\phi}HK16$ among $P1{\beta}$ subtype phages. Antibody $Ab{\phi}HK23$ of $P1{\gamma}$ subtype neutralized all phages of $P1{\beta}$ subtype as well as the phage ${\phi}HK23$, showing the widest phage-inactivation range. When the structural-similarity studies of phages were investigated by using phage antibodies, closeness obtained by phylogenetic analysis of 16S rRNA genes of pathogenic strains were quite different from that of polyclonal antibody-specific structural similarity of phage capsid proteins. In conclusion, there is weak correlation between the host strain specificity of bacteriophage and its capsid structural similarity measured by phage antibodies.

• Microbiology and Biotechnology Letters
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• v.25 no.1
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• pp.96-101
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• 1997

Among the lactic flora responsible for the development of acidity and characteristic flavor of Kimchi which is a traditional fermented Chiness cabbage. Homofermentative Lactobacillus plantarum is rod-shaped and to be known to ewert major role during later fermentation period. Once this strain establishes main flaora in the Kimchi fermentation process, it gives rise to excess acid production to reduce the taste and quality of Kimchi during storage. As a primary work to increase the keeping quality using virulent Lactobaillus plantarum bacteriophages, it was isolated sucessfully from collected Kimchi samples and their characteristics were studied. The new isolated phage, named SC 921, adsorbed to its host without Ca$^{2+}$, and nearly eliminated at 60$\circ $C of heat treatment for 5 min. This phages were atable at pH 4~ 10 but inactivated below pH 3.0 or pH 11.0 above. The latent period, rise period, and burst size of this phage was 100 min, 120 min, 31$\pm $2pfu/ml, respectively. Electron micrograph showed the phages particles were unusually oval feature of head (dia 80~ 120 nm) without contractile tail.

• Journal of Microbiology and Biotechnology
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• v.26 no.9
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• pp.1629-1635
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• 2016

A novel bacteriophage, PBES 02, infecting Cronobacter sakazakii was isolated and characterized. It has a spherical head of 90 nm in diameter and a tail of 130 nm in length, and belongs to Myoviridae as observed under a transmission electron microscope. The major virion protein appears to be 38 kilodaltons (kDa) in size. The latent period of PBES 02 is 30 min and the burst size is 250. Infectivity of the phage remained intact after exposure to temperatures ranging from 4℃ to 55℃ for 1 h. It was also stable after exposure to pHs ranging from 6 to 10 for 1 h. The phage effectively removed contaminating Cronobacter sakazakii from broth infant formula. PBES 02 has a double-stranded DNA genome of 149,732 bases. Its GC ratio is 50.7%. Sequence analysis revealed that PBES 02 has 299 open reading frames (ORFs) and 14 tRNA genes. Thirty-nine ORFs were annotated, including 24 related to replication and regulation functions, 10 related to structural proteins, and 5 related to DNA packaging. The genome of PBES 02 is closely related to that of two other C. sakazakii phages, CR3 and CR8. Comparison of DNA sequences of genes encoding the major capsid protein revealed a wide geographical distribution of related phages over Asia, Europe, and America.