• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.262-270
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• 2007

Of the twelve lytic bacteriophages recovered from five different fermenting cucumber tanks that were inoculated with Pediococcus sp. LA0281, a lytic phage, ${\phi}ps05$, was characterized in the present study. The plaques were mostly clear and round-shaped on the lawn of starter strain, indicating lytic phage. Overall appearance indicated that it belongs to the Siphoviridae family or Bradley's group B1, with a small isometric head and a flexible noncontractile tail with swollen base plate. The average size was found to be 51.2 nm in head diameter and 11.6 nm wide ${\times}$ 129.6 nm long for the tail. The single-step growth kinetics curve showed that the eclipse and the latent period were 29 min and 34 min, respectively, and an average burst size was calculated to be 12 particles per infective center. The optimum proliferating temperature ($35^{\circ}C$) was slightly lower than that of cell growth ($35\;to\;40^{\circ}C$). The structural proteins revealed by SDS-PAGE consisted of one main protein of 33 kDa and three minor proteins of 85, 58, and 52 kDa. The phage genome was a linear double-stranded DNA without cohesive ends. Based on the single and double digestion patterns obtained by EcoRI, HindIII, and SalI, the physical map was constructed. The overall size of the phage genome was estimated to be 24.1 kb. The present report describes the presence of a lytic phage active against a commercial starter culture Pediococcus sp. LA0281 in cucumber fermentation, and a preliminary study characterizes the phage on bacterial successions in the process of starter-added cucumber fermentation.

• 식물병연구
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• 제17권1호
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• pp.90-94
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• 2011

Xanthomonas oryzae pv. oryzae에 의해서 발생하는 벼 흰잎마름병은 세계적으로 벼 재배지역에서 큰 피해를 주고 있다. 그러나, 이 병의 효과적인 방제 수단은 적은 실정이다. 본 연구에서는 국내 벼 재배지의 관개수에서 분리한 Xoo 파지를 이용하여 흰잎마름병을 생물적으로 방제하고자 하였다. 먼저 포장에서의 사용가능성을 확인하기 위하여 pH, 온도, 자외선에 대한 Xoo 파지의 안정성을 조사하였다. Xoo 파지는 pH 5-pH 10 사이에서는 안정적이었고, $50^{\circ}C$에서 1시간 이상 처리시 대부분 불활성화 되었다. 자외선을 조사하였을 때는 약 2분간의 노출에서도 불활성화 되었다. 벼의 유묘에 살포한 파지는 세균이 없이도 7일간 벼 잎에서 활성을 유지하였고, 스킴 밀크에 혼합하여 살포하면 PBS 완충액에 넣어 살포한 것 보다 10배 이상의 생존율을 보였다. 그러나 이러한 생존율의 증가에도 불구하고 포장에서의 방제효과는 저조하였다. 하지만 파지 혼합액을 Tecloftalam WP, ASM 등과 혼합하여 동시에 살포하면 방제효과가 월등히 증가하였다. 이상의 결과는 Xoo 파지가 흰잎마름병의 방제에 사용될 수 있으며, 약제의 사용 빈도나 농도를 줄여 줄 수 있을 것임을 의미한다.

• 한국미생물·생명공학회지
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• 제25권1호
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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.

• BMB Reports
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• 제40권4호
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• pp.539-546
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• 2007

The lysozymes encoded by bacteriophage T7 and K11 are both bifunctional enzymes sharing an extensive sequence homology (75%). The constructions of chimeric lysozymes were carried out by swapping the N-terminal and C-terminal domains between phage T7 and K11 lysozymes. This technique generated two chimeras, T7K11-lysozyme (N-terminal T7 domain and C-terminal K11 domain) and K11T7-lysozyme (N-terminal K11 domain and C-terminal T7 domain), which are both enzymatically active. The amidase activity of T7K11-lysozyme is comparable with the parental enzymes while K11T7-lysozyme exhibits an activity that is approximately 45% greater than the wild-type lysozymes. Moreover, these chimeric constructs have optimum pH of 7.2-7.4 similar to the parental lysozymes but exhibit greater thermal stabilities. On the other hand, the chimeras inhibit transcription comparable with the parental lysozymes depending on the source of their N-terminals. Taken together, our results indicated that domain swapping technique localizes the N-terminal region as the domain responsible for the transcription inhibition specificity of the wild type T7 and K11 lysozymes. Furthermore, we were able to develop a simple and rapid purification scheme in purifying both the wild-type and chimeric lysozymes.

• 대한의생명과학회지
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• 제10권2호
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• pp.65-73
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• 2004

I report that single-stranded antisense as a part of large circular (LC-) genomic DNA of recombinant M13 phage exhibits enhanced stability, sequence specific antisense activity, and no need for target site search. A cDNA fragment (708 bp) of rat TNF-$\alpha$ was inserted into a phagemid vector, and TNF-$\alpha$ antisense molecules (TNF$\alpha$-LCAS) were produced as single-stranded circular DNA. When introduced into a rat monocyte/macrophage cell line, WRT7/P2, TNF$\alpha$-LCAS was able to ablate LPS-induced TNF-$\alpha$ mRNA to completion. The antisense effect of TNF$\alpha$-LCAS was shown to be sequence-specific because expressions of three control genes ($\beta$-actin, GAPDH and IL-1$\beta$) were not significantly altered by the antisense treatment. Further, TNF$\alpha$-LCAS was found to be highly efficacious as only 0.1 $\mu$g (0.24 nM) of TNF$\alpha$-LCAS was sufficient to block TNF-$\alpha$ expression in 1$\times10^5$ WRT7/P2 cells. I have also observed specific antisense activity in reduction of NF-$\kappa$B gene expression. The results suggest that an antisense sequence as a part of single-stranded circular genomic DNA has a specific antisense activity.

• KSBB Journal
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• 제10권5호
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• pp.533-539
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• 1995

재조합 대장균에 의해 생합성된 PHB를 분리정제하기 위하여 박테리오파아지의 세포파괴작용을 이용하는 방법의 가능성에 대해 알아 보았다. 먼저, $cI_{857}$ 유전자를 지난 박테리오파아지 A를 대장균에 감염시킨 후 lysogen, XLl-Blue($\lambda$HL1)를 선별하고, 이 균주에 PHA 생합성 플라스미드를 도입시켜 원하는 균주인 XLI-Blue($\lambda$HL1, pSYLl05)를 만들였다. 숙주인 XLl-Blue, 열적유도에 의해 세포파괴가 가능한 XLl-Blue($\lambda$HL1), 그리고 세포파괴와 PHB 생합성이 모두 가능한 XLl-Blue(AHL1, pSYL105) 에 대하여 여러 가지 조건에서의 실험결과를 비교.검토하였다. XLI-Blue($\lambda$IL1, pSYLl05)의 경우 대수기에서는 열적유도만으로 세포파괴를 효과적으로 야기할 수 있었으나 PHB가 축적되는 정지기에서는 열척유도만으로 세포파괴를 일으킬 수 없었다. 세포 파괴를 보다 용이하게 하기 위하여 열적유도 빛 2% (v/v)의 chloroform을 사용하는 화학적용균을 병행 하였는데, 이 경우 세포파괴가 효과적으로 일어남을 관찰할 수 있었다.

• 한국포장학회지
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• 제30권1호
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• pp.15-21
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• 2024

The edible coating system, consisting of gelatin and bacteriophages, has been developed to enhance the microbial safety of wild vegetables. Newly isolated Escherichia coli phage EP and Staphylococcus aureus phage SP were loaded into the gelatin coating solutions. The phages remained significantly stable for up to 3 days, respectively, and exhibited rapid antibacterial capacity within 2 h of coating application (p < 0.05). The developed coating was applied to bracken and exhibited antibacterial efficacy against E. coli and S. aureus within 6 h (1.9-log CFU/mL and 1.5-log CFU/mL). Furthermore, the coated bracken significantly prevented weight loss and maintained firmness for 10 days (p < 0.05). Consequently, gelatin-based edible coatings containing phages have the potential as an antibacterial packaging strategy.

• Animal Bioscience
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• 제35권11호
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• pp.1752-1759
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• 2022

Objective: Effects of direct-fed Enterococcus faecium plus bacteriophages (EF-BP) were investigated as potential substitutes for pharmacological ZnO for weanling pigs. Methods: Dietary treatments were supplementations to a basal diet with none (NC), 3,000-ppm ZnO (PC), 1×10¹⁰ colony-forming units of E. faecium plus 1×10⁸ plaque-forming units (PFU) of anti-Salmonella typhimurium bacteriophages (ST) or 1×10⁶ PFU of each of anti-enterotoxigenic Escherichia coli K88 (F4)-, K99 (F5)-, and F18-type bacteriophages (EC) per kg diet. In Exp 1, twenty-eight 21-day-old crossbred weanling pigs were individually fed one of the experimental diets for 14 days and euthanized for histological examination on intestinal mucosal morphology. In Exp 2, 128 crossbred weanling pigs aged 24 days were group-fed the same experimental diets in 16 pens of 8 piglets on a farm with a high incidence of post-weaning diarrhea. Results: None of the diarrheal score or fecal consistency score (FCS), average daily gain (ADG), gain: feed ratio, structural variables of the intestinal villus, and goblet cell density, differed between the EF-BP (ST+EC) and NC groups, between EF-BP and PC, or between ST and EC, with the exception of greater gain: feed for EF-BP than for PC (p<0.05) during days 7 to 14 (Exp 1). In Exp 2, ADG was less for EF-BP vs PC during days 0 to 7 and greater for EF-BP vs NC during days 7 to 14. FCS peaked on day 7 and declined by day 14. Moreover, FCS was less for EF-BP vs NC, did not differ between EF-BP and PC, and tended to be greater for ST vs EC (p = 0.099). Collectively, EF-BP was comparable to or slightly less effective than PC in alleviating diarrhea and growth check of the weanling pigs, with ST almost as effective as PC, when they were group-fed. Conclusion: The E. faecium-bacteriophage recipe, especially E. faecium-anti-S. typhimurium, is promising as a potential substitute for pharmacological ZnO.

• 한국미생물·생명공학회지
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• 제10권4호
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• pp.253-258
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• 1982

본 실험의 결과를 요약해 보면 인산염의 종류와 농도에 따라서 유산균의 증식과 phage 감염후의 유산균 증식에 미치는 영향을 검토하였다. 특히, phage 증식억제를 나타내는 물질은 sodium metaphosphate, sodium pyrophosphate, sodium phosphate (dibasic), potassium phosphate(dibasic)으로써 각각 0.2∼0.4%, 0.1∼0.2%, 0.2∼0.3%, 0.2∼0.35%에서 phage 감염 후 유산균의 증식을 가능하게 하였다. 그리고 phage 감염 후에도 증식한 유산균 배양액을 새로운 배지에 1,2차 계대 하였을 때, 역시 인산염의 종류와 농도에 따라 배양액에 존재하는 phage에 의해 유산균의 증식에 큰 영향을 미쳤다. 특히 sodium pyrophosphate의 경우, 0.17%이상에서는 1차 계대 후에도 phage의 증식이 억제되어 유산균이 증식하기 때문에 starter 배양액용 P.R.M제조에 이용이 가능한 것으로 보여 진다.

• Food Science and Biotechnology
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• 제17권6호
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• pp.1221-1227
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• 2008

The application of the cellulase gene (celA) as a selection marker of food-grade integration system was investigated in Lactobacillus (Lb.) casei, Lactococcus lactis, and Leuconostoc (Leu.) mesenteroides. The 6.0-kb vector pOC13 containing celA from Clostridium thermocellum with an integrase gene and a phage attachment site originating from bacteriophage A2 was used for site-specific recombination into chromosomal DNA of lactic acid bacteria (LAB). pOC13 was also equipped with a broad host range plus replication origin from the lactococcal plasmid pWV01, and a controllable promoter of nisA ($P_{nisA}$) for the production of foreign proteins. pOC13 was integrated successfully into Lb. casei EM116, and pOC13 integrants were easily detectable by the formation of halo zone on plates containing cellulose. Recombinant Lb. casei EM 116::pOC13 maintained these traits in the absence of selection pressure during 100 generations. pOC13 was integrated into the chromosome of L. lactis and Leu. mesenteroides, and celA acted as an efficient selection marker. These results show that celA can be used as a food-grade selection marker, and that the new integrative vector could be used for the production of foreign proteins in LAB.