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Characteristics and Lytic Activity of Phage-Derived Peptidoglycan Hydrolase, LysSAP8, as a Potent Alternative Biocontrol Agent for Staphylococcus aureus

  • Yu, Jun-Hyeok;Lim, Jeong-A;Chang, Hyun-Joo;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1916-1924
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    • 2019
  • Outbreaks of staphylococcal food poisoning (SFP) causing serious human diseases and economic losses have been reported globally. Furthermore, the spread of Staphylococcus aureus with increased resistance to multiple antimicrobial agents has become a major concern in the food industries and medicine. Here, we isolated an endolysin LysSAP8, as one of the peptidoglycan hydrolases, derived from the bacteriophage SAP8 infecting S. aureus. This endolysin was tagged with a 6×His at the C-terminal of the target protein and purified using affinity chromatography. LysSAP8 demonstrated lytic activity against a broad spectrum of bacteria, which included a majority of the staphylococcal strains tested in this study as well as the methicillin-resistant S. aureus (MRSA); however, no such activity was observed against other gram-positive or gram-negative bacteria. Additionally, LysSAP8 could maintain bactericidal activity until 0.1 nM working concentration and after heat treatment at 37℃ for 30 min. The ability of LysSAP8 to lyse cells under varying conditions of temperature (4-43℃), pH (3-9), and NaCl concentrations (0-1,000 mM), and divalent metal ions (Ca2+, Co2+, Cu2+, Mg2+, Mn2+, Hg2+, and Zn2+) was examined. At the optimized condition, LysSAP8 could disrupt approximately 3.46 log CFU/ml of the planktonic cells in their exponential phase of growth within 30 min. In this study, we have suggested that LysSAP8 could be a potent alternative as a biocontrol agent that can be used to combat MRSA.

Purification and Characteristics of Endo-Polygalacturonase from Korean Tomato (한국산 토마토의 Endo-Polygalacturonase 정제 및 성질)

  • Choi, Cheong;Cho, Young-Je;Son, Gyu-Mok
    • Applied Biological Chemistry
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    • v.33 no.1
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    • pp.73-78
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    • 1990
  • Endo-polygalacturonase was purified from tomato, Lycopersicon esculentum L. The purification procedures included gel filtration on Sephadex G-150 and DEAE-cellulose ion exchange chromatography. Yield of the enzyme purification was 12.74 %. Purified enzyme was confirmed as a active single band by the SDS-polyacrylamide gel electrophoresis. When the purified enzyme was applied to SDS-PAGE, the molecular weight was estimated about 50,000. The optimum pH for the enzyme activity was 5.0 and the range of its stability to the pH was 4.0 to 5.0. The optimum temperature was $50^{\circ}C$, while the enzyme was abruptly inactivated above $50^{\circ}C$. From the result of the study on the effects of metals ion, it was found that $Ag^+$, $Zn^{++}$ and $Mg^{++}$ increased on the enzyme activity. In contrast, $Ba^{++}$, $Hg^{++}$, $Pb^{++}$, $Ca^{++}$, $Mn{++}$, $Cu^{++}$, $Fe^{+++}$, $Na^+$ and $K^+$ decreased it. the reaction catalyzed by this enzyme followed typical Michaelis-Menten kinetics with the Km value of $1.43{\times}10^{-1}\;mol/l$.

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Characterization of a Novel Fibrinolytic Enzyme, BsfA, from Bacillus subtilis ZA400 in Kimchi Reveals Its Pertinence to Thrombosis Treatment

  • Ahn, Min-Ju;Ku, Hye-Jin;Lee, Se-Hui;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2090-2099
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    • 2015
  • Recently, the cardiovascular disease has been widely problematic in humans probably due to fibrin formation via the unbalanced Western style diet. Although direct (human plasmin) and indirect methods (plasminogen activators) have been available, bacterial enzyme methods have been studied because of their cheap and mass production. To detect a novel bacterial fibrinolytic enzyme, 111 bacterial strains with fibrinolytic activity were selected from kimchi. Among them, 14 strains were selected because of their stronger activity than 0.02 U of plasmin. Their 16S rRNA sequence analysis revealed that they belong to Bacillus, Leuconostoc, Propionibacterium, Weissella, Staphylococcus, and Bifidobacterium. The strain B. subtilis ZA400, with the highest fibrinolytic activity, was selected and the gene encoding fibrinolytic enzyme (bsfA) was cloned and expressed in the E. coli overexpression system. The purified enzyme was analyzed with SDS-PAGE, western blot, and MALDI-TOF analyses, showing to be 28.4 kDa. Subsequently, the BsfA was characterized to be stable under various stress conditions such as temperature (4-40oC), metal ions (Mn2+, Ca2+, K2+, and Mg2+), and inhibitors (EDTA and SDS), suggesting that BsfA could be a good candidate for development of a novel fibrinolytic enzyme for thrombosis treatment and may even be useful as a new bacterial starter for manufacturing functional fermented foods.

다슬기로부터 칼슘락테이트의 제조와 품질특성

  • 김순동;이예경;이명예;장경호
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.04a
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    • pp.122-122
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    • 2003
  • 다슬기의 분말(PSB)과 그 회화분(ASB)으로부터 체내 흡수력이 높은 젖산칼슘의 제조조건과 색상, 용해도 및 관능적 품질을 조사하였다. PSB로부터 제조한 젖산칼슘(PSB-CL)의 PSB 및 젖산 100 mL에 대한 수율은 젖산농도가 10%일 때는 300% 및 15 g, 20%일 때는 260% 및 20 g이었으며, ASB로 제조한 젖산칼슘(ASB-CL)의 ASB 및 젖산 100 mL에 대한 수율은 10% 젖산에서는 400% 및 60 g, 20% 젖산에서는 329% 및 66 g으로 원료량을 기준으로 하였을 때는 다같이 젖산농도의 증가에 따라 감소하였으나 젖산의 부피를 기준으로 하였을 때는 젖산농도의 증가에 따라 증가하였다. Dehydated PSB-CL 및 ASB-CL 제조의 적정온도와 시간은 10$0^{\circ}C$에서는 각각 4 및 5시간, 12$0^{\circ}C$에서는 3 및 4시간, 15$0^{\circ}C$에서는 1 및 2시간으로 ASB-LA의 경우가 짧았다. IR 및 H-NMR spectrum의 분석결과 PBS-LA와 ASB-LA의 구조는 Ca($CH_3$CHOH$CO_2$)$_2$임이 확인되었다. 무수 PSB-CL 및 ASB-CL의 칼슘함량은 각각 15.4%(w/w)와 17.3%(w/w)로 이론 값의 각각 84.2%와 94.5%를 나타내었으며, 미량의 Fe, Na, Mn Zn을 함유하였다. PBS-CL와 ASB-CL의 색상은 각각 연한 황색과 연녹색을 지닌 백색을 띠었다. pH 3~8로 조정한 증류수에서 PSB-CL과 ASB-CL의 평균 용해도는 각각 5.43 g/100 mL 및 6.11 g/100 mL로 standard CL의 4.74 g/mL에 비하여 높았다. 국간장을 제외한 대부분의 액체식품(3% 소금물, 소주, 진간장, 국간장, 포도주스 및 오렌지주스)에서의 용해도는 PSB-CL(3.14~5.03 g/100 mL)과 ASB-CL(4.69~6.05 g/100 mL)이 standard CL(2.94~5.84 g/100 mL)에 비하여 높았다. 관능검사 결과, ASB-CL은 신맛이 낮아 사용범위가 높은 것으로 평가되었으며 PSB-CL는 쓴맛은 높으나 떫은맛이 낮고 구수한 맛이 강하여 식품에의 응용이 기대된다.

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Effects of ZnO on the Piezoelectric Properties of PMS-PZT Ceramics (PMS-PZT 세라믹스의 압전특성에 미치는 ZnO의 영향)

  • Son Y.-J.;Hwang D.-Y.;Kim J.-C.;Cho K.-W.;Kim Y.-M.;Ur S.-C.;Kim I.-H.
    • Korean Journal of Materials Research
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    • v.14 no.11
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    • pp.764-768
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    • 2004
  • Perovskite Pb(Mn_{1/3}Sbu_{2/3})O_2-Pb(Zr,Ti)O_3\;(PMS-PZT) was prepared and ZnO doping effects on its piezoelectric properties were investigated. Pyrochlore phase was not identified in the PMS-PZT ceramics with $0\sim5\;mol\%$ ZnO sintered at $1100^{\circ}C$ for 2 hrs, and maximum sintered density of $7.92 g/cm^3$ was obtained. Piezoelectric charge constant and voltage constant increased to $359{\times}10^{-12}\;C/N\;and\;22.5{\times}10^{-13}\;Vm/N$, respectively, with increasing ZnO content. Mechanical quality factor reduced considerably with increasing ZnO content. When the ZnO content was 3 $mol\%$, electromechanical coupling factor and relative dielectric constant showed maximum values of $56\%$ and 1727, respectively. This should be evaluated by complicated variations of sintered density, tetragonality of lattice, grain size, and A-site vacancy generated by ZnO addition and $Zn^{2+}$ substitution.

Investigation of Micromorphological Characteristics of Acupuncture Needle Tip Using SEM-EDX (SEM-EDX를 이용한 침 끝의 미세 부착물의 조성에 대한 관찰)

  • Jang, In-Soo;Son, Dong-Hyuk;Song, Ho-Seop;Lee, In-Hwan;Park, Jong-Bae
    • Journal of Acupuncture Research
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    • v.22 no.6
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    • pp.135-140
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    • 2005
  • Objectives : There have been several studies about the quality of acupuncture needle tip recently. We have investigated the condition of the tip of the acupuncture needles in the last studies. In the former studies, we discovered the metallic scuff, lumps and irregularities of the acupuncture needle tips under the microscope. But, no information was available on those foreign materials' identity. Methods : We have selected 200 needles of 1000 pieces from several companies by randomized methods. And we observed the tip of the 6 needles selected finally at ${\times}1000\;or\;{\times}3000$ magnification and analyzed the components of the metallic scuff, lumps and irregularities of the needle tips with a SEM-EDX analyser. Results : We found that the identity of the metallic scuff, lumps and irregularities of the needle tips were metallic materials and silicon. For example, A point was composed of Fe(69.78%), Cr(17.71%), Ni(8.11%), Zn(2.04%), Si(1.23%), Mn(1.12%), and B point was composed of Si(66.40%), Fe(26.76%), Cr(6.84%). Conclusion : The results of this study confirm that there is a real possibility of the remaining of metallic materials and silicon in body of patient, after acupuncture treatment. Therefore, it is necessary to intensify our efforts to make needles of good quality and to concentrate on manufacturing process of acupuncture needles completely to be free from danger in acupuncture treatment.

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Investigation on Contents of Heavy Metals and Parasites of the Dropwort(Oenanthe javanica) cultivated in Chollanamdo (전라남도내 미나리의 중금속 함량 및 기생충에 대한 조사)

  • Heo, Nam-Chil;Kim, Choong-Mo;Park, Sook;Na, Hwan-Sik
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.970-973
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    • 2000
  • Heavy metal contents and parasite egg examination for the safety evaluation of dropwort(Oenanthe javanica) from the inside of Chonnam province(5 regions) and suburbs of Kwang-ju(1 region) were investigated. The contents of heavy metals(Hg, Pb, As, Cd, Mn, Zn, Cu) were determined by the Atomic Absorption Spectrophotometer and parasite eggs were examined to various internalparasites(F. hepatica, C. sinesis, G. lamblia, Cryptosporidium, A. lumbricoides and A. duodenale). First, Hg and As were detected in trace level and $0.003{\sim}0.01\;ppm$. Pb was not detected in all samples and Cd contents of dropworts were detected $0.014{\sim}0.087\;ppm$. The contents of 4 kinds of heavy metals(Hg, As, Pb and Cd) in dropworts cultivated in various regions(6 regions) were not detected or trace levels. Therefore, dropworts cultivated in Chonnam province and suburbs of Kwang-ju were free from contamination of heavy metals. Moreover, the result of parasite egg examination(Trematoda, Protozoa and Nematoda) was not detected in all samples.

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Ionic and Elemental Compositions of PM2.5 at the 1,100 m-Highland of Mt. Hallasan in Jeju Island (한라산 1,100 m 지역의 대기 중 PM2.5에 함유된 이온 및 원소 성분의 조성특성)

  • Lee, Ki-Ho;Hu, Chul-Goo
    • Journal of Environmental Science International
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    • v.25 no.6
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    • pp.865-875
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    • 2016
  • In this study, mass concentrations and chemical compositions of $PM_{2.5}$, including water-soluble ions and elements were determined at the 1,100 m-highland of Mt. Hallasan in Jeju Island across four seasons from August 2013 to August 2014. The average mass concentration of $PM_{2.5}$ was $12.5{\pm}8.41{\mu}g/m^3$ with 45.8% of the contribution from eight water-soluble ionic species. Three ionic species ($SO{_4}^{2-}$, $NH{_4}^+$, and $NO{_3}^-$) comprised 96.2% of the total concentration of ions contained in $PM_{2.5}$ and were the dominant ions, accounting for 43.5% of the $PM_{2.5}$ mass at Mt. Hallasan. On the basis of the mass concentration level, seasonal variation, enrichment factor, and relationship among elements, we can presume that Mg, K, Ca, Mn, Fe, Co, Sr, Ba, Nd, and Dy originated mainly from crust or soil and that V, Cr, Ni, Cu, Zn, As, Cd, and Pb were significantly enriched in $PM_{2.5}$ owing to the effects of the anthropogenic emissions. These results and the local distribution of emission sources and topographic characteristics near this sampling site suggest that the compositions of $PM_{2.5}$ collected at the 1100 m-highland of Mt. Hallasan were largely influenced by inflow from outside of Jeju Island.

Volatile Sulfur Compounds, Proximate Components, Minerals, Vitamin C Content and Sensory Characteristics of the Juices of Kale and Broccoli Leaves (케일 및 브로콜리잎즙의 함황 향기성분, 일반성분, 무기질, Vitamin C 함량 및 관능적 특성)

  • 김미리;김진희;위대성;나종현;석대은
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.6
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    • pp.1201-1207
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    • 1999
  • To utilize the leaves of broccoli, the shape of which was similar with that of kale leaves, as a vegetable juice, the extracts from two types of vegetable leaves were subjected to the analyses of proximate com ponents, volatile sulfur compounds, vitamin C and minerals. The sensory evaluation of the juices, prepared from kale and broccoli leaves, were performed by duo trio test and scoring test. Among eight varieties of broccoli, four varieties('Pilgrim', 'Greenbelt', 'Salinas' and 'Shasta')were not significantly different from kale(p<0.05), based on the duo trio sensory test. Sulfur and nitrogen compounds present in the dichloromethane extracts of kale and broccoli leaves 'Pilgrim' were identified as 3 butenyl, allyl, butyl and 4 methylsulfinylbutyl isothiocyanate(sulforaphane), 4,5 epithiovalero, 4,5 epithio 3 hydroxyvalero, benzenepropane, 3 hydroxy 4 propene, 4 methylthio butane, 3 hydroxy 3 phenylpropane, 5 methy lsulfinylpentane, 4 methoxyphenyl 3 hydroxypropane nitrile and dimethyl trisulfide by GC/MSD analysis. Proximate components were observed to be relatively similar between kale and broccoli leaves. Vitamin C and sugar content were higher in broccoli leaves(125∼180mg% and 8∼12°Brix) than in kale(101mg% and 7oBrix). In mineral analysis by AA and ICP Mass, 'Pilgrim' showed a higher content of K, Fe, Mn, Zn and Se. In separate experiment, sensory scores of bitter taste and astringent taste were lower and scores of over all taste and over all acceptability, higher in broccoli('1243') juice than those in kale juice.

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Characteristics and Quantity of Slurry Produced by Swine Slurry Farms (슬러리 돈사에서의 슬러리 발생량 및 이화학적 특성)

  • Kwag, J.H.;Choi, H.C.;Choi, D.Y.;Kang, H.S.;Park, C.H.;Han, J.D.;Jeon, B.S.;Kim, H.H.
    • Journal of Animal Environmental Science
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    • v.8 no.2
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    • pp.111-114
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    • 2002
  • This study was conducted to determine the volume of pig slurry productinn and the characteristics from 4 swine farms. For the composition of pig slurry produced, contents of N, $P_2O_5$ and $K_2O$, were 0.13, 0.25 and 0.13% in slurry, respectively. Water pollutant concentration in slurry of swine farms, $BOD_5$, $COD_MN$, SS, T-N and T-P, was $24,047mg/{\ell}$, $30,232mg/{\ell}$, $36,833mg/{\ell}$, $2,805mg/{\ell}$, $465mg/{\ell}$, respectively. The average volume of pig slurry was 6.30 ${\ell}/head/day$ and 6.32 ${\ell}$ in spring, 6.69 ${\ell}$ in summer, 6.09 ${\ell}$ in autumn, and 6.12 ${\ell}$ in winter. The average moisture content of slurry was 95.8%. The composition of slurry produced by pig farms.

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