• Title/Summary/Keyword: pH-sensitive

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Microsomal Proton Transport Activity Measured by Quinacrine Fluorescence from Tomato Roots (Quinacrine 형광을 이용한 토마토 뿌리조직 마이크로솜의 수소이온이동 활성측정)

  • Shin, Dae-Seop;Cho, Kwang-Hyun;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.45 no.2
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    • pp.53-58
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    • 2002
  • Quinacrine, a pH-sensitive fluorescence probe, which exists either as an unprotonated fluorescence form or a protonated noufluorescence form, can be used to measure the proton transport activity of $H^+-ATPase$. Quinacrine was used to determine the optimal conditions for measuring the activity of microsomal $H^+-ATPase$ prepared from the roots of tomato plants. The amount of quinacrine fluorescence quenching obtained at $0.43{\mu}g/{\mu}l$ of microsomal protein concentration was 25-26%, which shows that the enzyme activity of 100 nmol/min decreases 10% of quinacrine fluorescence. Maximal fluorescence quenching was obtained at pH 7.0-7.2 and 2 mM $Mg^{2+}$ Because the activity of microsomal $H^+-ATPase$ is also maximal at these conditions, the quinacrine fluorescence well represents the activity of $H^+-ATPase$. Vanadate and $NO_3-$, specific inhibitors of plasma and vacuolar $H^+-ATPases$, respectively, were successfully applied to inhibit the quinacrine fluorescence quenching mediated by the corresponding $H^+-ATPases$. These results imply that quinacrine is a useful tool for measuring the proton transport activities of microsomes obtained from the root tissue of tomato plants.

Isolation and Characterization of an Anti-listerial Bacteriocin from Leuconostoc lactis SD501

  • Hwang, In-Chan;Oh, Ju Kyoung;Kim, Sang Hoon;Oh, Sejong;Kang, Dae-Kyung
    • Food Science of Animal Resources
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    • v.38 no.5
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    • pp.1008-1018
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    • 2018
  • Although bacteriocins with anti-listerial activity have been isolated from a wide variety of lactic acid bacteria, little is known about those from Leuconostoc lactis, a heterofermentative bacterium that produces diacetyl and exopolysaccharides in dairy foods. In this study, an anti-listerial bacteriocin was isolated from Leuc. lactis SD501 and characterized. It was particularly potent against Listeria monocytogenes and also inhibited Enterococcus faecalis. Anti-listerial activity reached a maximum during the early stationary phase and then decreased gradually. The anti-listerial substance was sensitive to proteinase K and ${\alpha}$-chymotrypsin, confirming its proteinaceous nature. Its activity remained stable at pH values ranging from 1 to 10. In addition, it was strongly resistant to high temperatures, retaining its activity even after incubation for 15 min at $121^{\circ}C$. The apparent molecular mass of the partially purified anti-listerial bacteriocin was approximately 7 kDa. The characteristics of the SD501 bacteriocin, including its small molecular size (<10 kDa), strong anti-listerial activity, wide pH stability and good thermostability, indicate its classification as a Class IIa bacteriocin.

Determination of Dioxin-like Components in the School Waste Incinerator Residues by EROD-microbioassay (EROD-microbioassay에 의한 학교 소각로 잔재 중 다이옥신 유사물질의 측정)

  • 정규혁;오승민;윤완진
    • Journal of Environmental Health Sciences
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    • v.26 no.3
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    • pp.11-17
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    • 2000
  • There are among the most relevant toxic emissions from incinerators such as polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), dioxin-like polychlorinated biphenyls (PCBs). Induction of cytochrome P4501A1 catalyzed 7-ethoxyresorufin O-deethylase(EROD) activity in mammalian cell culture(EROD bioassay) is thought to be a selective and sensitive parameter used for the quantification of dioxin-like components. In this study, the toxic emissions from several school waste incinerators were evaluated by determination of CYPIA catalytic activity and cytotoxicity using cell culture microbioassay. The incinerator residue and soil samples were collected from the schools located in Kyunggi province from April to June 1999. The samples were extracted in a Soxhlet apparatus using toluene for 20 hours. In order to clean-up, concentrated crude extracts were applied to basic alumina column. The EROD activities of extracts in the H4IIE cells were from 1.91$\pm$0.32 ng-TEQ/g to 24.54$\pm$3.48 ng-TEQ/g of biochemical-TEQ value. In soil samples, CYP1A catalytic activity was 0.09~0.64 ng-TEQ/g. EROD bioassay, seems to be a useful short-term bioassay when information about the biological response of complex environmental samples is needed. Although further study is needed, these results indicate that the potent toxic emissions are produced from school waste semi-incinerators.

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Isolation and Characterization of Four Carboxypeptidases in Canavalia lineata Cotyledons

  • Yang, Jong-Moon;Rhew, Tae-Hyong;Koh, Suck-Chan;Kwon, Young-Myung
    • BMB Reports
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    • v.28 no.5
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    • pp.451-457
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    • 1995
  • Four carboxypeptidases, CP1, CP2, CP3, and CP4 were isolated from the cotyledons of germinating seedlings of Canavalia lineata by sequential chromatography on the following four columns: 1) CM-cellulose, 2) Sephacryl 5-300, 3) Procion red dye, and 4) Sephacryl S-200. A number of properties of the enzymes, such as substrate specificity, molecular weight, optimum pH, thermal stability, have been determined. Enzyme activities were measured using the Cbz(carbobenzoxy)-dipeptides containing phenylalanine at the penultimate position. The $K_m$ values of four carboxypeptidases for Cbz-Phe-Ala were 0.50, 0.65, 1.30, and 1.35 mM, respectively. The inhibition studies indicated that the four carboxypeptidases were all serine type. Each of the carboxypeptidases with molecular weights of 145, 114, 105, and 104 kDa, respectively, had the optimum enzyme activity at pH 5.0~6.0. And they were sensitive to high temperature.

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Characterization of a Bacteriocin Produced by Enterococcus sp. T7 Isolated from Humans

  • Moon, Hi-Seong;Jeong, Jong-Jin;Ji, Geun-Eog;Kim, Jong-Sang;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.507-513
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    • 2000
  • A bacteriocin-producing organism, Enterococcus sp. T7, was isolated from human fecal samples. Bacteriocin T7, named tentatively as the bacteriocin, was produced by Enterococcus sp. T7 and it inhibited some strains of Lactobacillus. Staphylococcus, Enterococcus, and Streptococcus, but not all the lactococci and gram-negative bacteria tested. Bacteriocin T7 inhibited the growth of Listeria monocytogenes Scott A, but the degree of inhibition was less than those for other sensitive gram-positive vacteria. Bacteriocin T7 in MRS broth started to produce at the middle of the exponential growth phase and the inhibitory activity reached its maximum level during the stationary growth phase. Bacteriocin T7 was stable against heat treatments, pH variations (pH 2-10), and exposure to organic solvents. The molecular weight of bacteriocin T7 was estimated to be 6.500 Da by SDS-PAGe. All these facts, including physico-chemical stabilities, small molecular size, and inhibition of Kisteria monocytogenes, indicate that bacteriocin T7 is likely to be a member of the class IIa bacteriocins.

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Isolation and Characterization of Pseudomonas putida N3 Degrading Naphthalene (Naphthalene을 분해하는 Pseudomonas putida N3의 분리 및 특성)

  • 고영희;하일호;배경숙
    • Microbiology and Biotechnology Letters
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    • v.16 no.3
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    • pp.199-204
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    • 1988
  • A strain capable of growth on naphthalene minimal medium was isolated from soil by selective enrichment culture and identified as Pseudomonas putida N3 according to its morphological and physiological characteristics. The optimum pH and temperature for growth of the isolate were 7.0 and 3$0^{\circ}C$, respectively. This strain was resistant to ampicillin, chloramphenicol, kanamycin and streptomycin but. sensitive to tetracycline and rifampicin. Of the naphthalene related compounds, 1, 5-dihydroxynaphthalene was more easily utilized than naphthalene due to its solubility. And catechol was degraded through meta-cleavage pathway. A 110 Kb plasmid which encodes for a single set of enzymes responsible for the degradation of naphthalene was obtained.

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Application of in situ Liquid Ferrate(VI) for 2-Bromophenol Removal (2-브로모페놀 제거를 위한 액상 페레이트 적용 연구)

  • Laksono, Fajar Budi;Kim, Il-Kyu
    • Journal of Korean Society of Water and Wastewater
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    • v.29 no.6
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    • pp.685-692
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    • 2015
  • The concern over the risk of environmental exposure to brominated phenols has been increased and has led the researchers to focus their attention on the study of bromophenol treatment. In this study, the effects of pH and ferrate(VI) dose on the degradation of 2-bromophenol were investigated. The results indicated that the oxidation of 2-bromophenol by liquid ferrate(VI) was found to be highly sensitive to the pH condition. Furthermore, the highest removal efficiency was observed at the neutral condition with the removal efficiency of 94.2%. In addition, experimental results showed that 2-bromophenol removal efficiency increased with increasing of ferrate dosage. Ferrate(VI) dose of 0.23 mM was sufficient to remove most of the 2-bromophenol with the efficiency of 99.73% and kapp value of $2982M^{-1}s^{-1}$. Seven compounds were identified as the intermediate products by the GC/MS analysis.

Adsorptive Stripping Voltammetry of Indium-Morin Complex (Indium-Morin 착물에 관한 흡착벗김전압전류법적 연구)

  • Se Chul Sohn;Tae Yoon Eom;Yeong Kyeong Ha;Ki-Suk Jung
    • Journal of the Korean Chemical Society
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    • v.35 no.5
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    • pp.506-511
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    • 1991
  • A sensitive adsorptive stripping voltammetric study was investigated on the complex of indium with morin at a hanging mercury drop electrode in 0.1 M acetate buffer (pH 3.20) solution. The adsorption phenomena were observed by differential-pulse voltammetry. The effects of various analytical conditions were discussed on the reduction peak current of the adsorbed complex on the surface of HMDE. Interferences by other trace metals and surfactant were also discussed. Detection limit was 2.6 nM of indium after 90 second deposition time, and the relative standard deviation (n = 7) at 4TEX>${\mu}g$/l indium was 2.0%.

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Development of a Virus Elution and Concentration Procedure for Detecting Norovirus in Oysters

  • Ha, Sook-Hee;Woo, Gun-Jo;Hwang, In-Gyun;Choi, Weon-Sang
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1150-1154
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    • 2009
  • Low levels of virus contamination and naturally occurring reverse transcription-polymerase chain reaction (RT-PCR) inhibitors restrain virus detection in oysters. A rapid and efficient oyster-processing procedure that can be used for sensitive virus detection in oysters was developed. Poliovirus type 1 Sabin strain was used to evaluate the efficacy of virus recovery. The procedure included (a) acid-adsorption and elution with buffers (0.25M glycine-0.14 M NaCl, pH 7.5; 0.25M threonine-0.14M NaCl, pH 7.5); (b) polyethylene glycol (PEG) precipitation; (c) resuspension in Tween 80/Tris solution and chloroform extraction; (d) the second PEG precipitation; (e) viral RNA extraction with TRIzol and isopropanol precipitation; and (f) RT-PCR combined with semi-nested PCR. The overall recovery of elution/concentration was 19.5% with poliovirus. The whole procedure usually takes 19 hr. The overall detection sensitivity was 4 RT-PCR units of genogroup I norovirus (NoV) and 6.4 RT-PCR units of genogroup II Nov/25 g of oysters initially seeded. The virus-detecting method developed in this study should facilitate the detection of low levels of NoV in oysters.

Development of Cell Entrapment Technology for the Improvement of Bifidobacterium Viability (Bifidobacterium의 생존력 증대를 위한 세포포집기술개발)

  • Park, Hui-Gyeong;Bae, Gi-Seong;Heo, Tae-Ryeon
    • KSBB Journal
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    • v.14 no.4
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    • pp.389-395
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    • 1999
  • Bifidobcterium spp. can provide human being with several beneficial physiological. Therefor, there has been a considerable interest in products Bifidobcterium spp. dietary supplements or as starter cultures for probiotic products that may assint in the improvement of health on the human. But indusrial applications have been limited because Bifidobcterium spp. are sensitive to acidic pH due to organic acid produced by themselves and various conditions. The objective of this study was to establish new method for improvement of Bifidobcterium viability by entrapment im calcium alginate beads. We have a plan to select the most suitable polymer through the comparison with acid tolerance oxygen tolerance and theological properties of polymer. Increase of the viable number of Bifidobcterium induced increasing acid tolerance and oxygen tolernce trough the development of entrapment technique. The 4%, 3030mm diameter) sodium alginate beads led to the best survivability under acid condition. Especially, addition of 6% mannitol, 6% glycerol or 6% sorbitol to the sodium alginate helped a beneficial effect on viability against acid, bile salt, hydrogen peroxide and cold strage. The number of viability of entrapeede cells by retreatment was 96 fold higher than non-entrapeed cells after 5 hours of storage under pH 3 acidic condition. These experimental data clearly demonstrate that a whole cell immobilization by entrapment in calcium alginate beads is an important survival mechanism enable to withstand environmental stresses as the acidic condition, hydrogen peroxide toxicity and frozen state.

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