• Title/Summary/Keyword: Stationary phase

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Isolation and characterization of Vitreoscilla mutant defective in catalase-peroxidase hydroperoxidase I

  • Kim, Hee-Jung;Moon, Ja-Young;Lee, John-Hwa;Park, Kie-In
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.291-304
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    • 2007
  • Mutants of an obligate aerobic bacterium, Vitreoscilla, that have deficiency in heat-labile catalase-peroxidase hydroperoxidase I (HPI) were created by EMS treatment. The catalase-peroxidase HPI-deficient mutant showed substantially lower peroxidase activity in exponential and mid-stationary phase compared with the wild type strain. In late stationary phase, the mutant exhibited no peroxidase activity. Peroxidase deficiency in the mutant was revealed by polyacrylamide gels stained for peroxidase activity. Characteristically, catalase levels in the mutant increased about 14- and 8-fold during growth in the exponential and stationary phases, respectively, compared to those in the wild type, suggesting a compensatory effect for protection from $H_2O_2$ toxicity. The mutant showed differences in physiology from the wild type: retardation in growth rate and decrease in oxygen consumption. Both the wild type and the catalase-peroxidase HPI-deficient mutant of Vitreoscilla had lower growth rates in media containing increasing $H_2O_2$ concentrations. However, the mutant exhibited an additionally decreased growth rate after 6 to 8 h of growth compared to the wild type. The wild type was resistent up to 20 mM $H_2O_2$, whereas the mutant was very sensitive to high concentrations of exogenous $H_2O_2$. Although elevated catalase levels would provide protection of the bacteria from the deleterious effect of $H_2O_2$, it did not appear to be complete. Cell-free extracts of the mutant showed decreased NADH oxidation rates and higher accumulation of $H_2O_2$ during this oxidation. These results may account for the impaired growth and earlier onset of death phase by the catalase-peroxidase HPI-deficient mutant of Vitreoscilla.

Identification and Partial Characterization of Lacticin SA72, a Bacteriocin Produced by Lactococcus lactis SA72 Isolated from Jeot-gal

  • Koo, Kyoung-Mo;Lee, Na-Kyoung;Hwang, Young-Il;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.488-495
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    • 2000
  • Strain SA72 was isolated from Jeot-gal and identified as producer of a bacteriocin, which showed some bactericidal activity against Lactobacillus delbrueckii ATCC 4797. Strain SA72 was tentatively identified as Lactococcus lactis according to the AOI test. Lactococcus lactis SA72 showed a broad spectrum of microorganisms, tested by the modified deferred method. The activity of lacticion SA72, named tentatively as a bacteriocin produced by Lactococcus lactis SA72, was detected during the mid-lon growth phase, reached a maximum during the early stationary phase, and then declined after the late stationary phase. Lacticin SA72 also showed a relatively broad spectrum of activity against non-pathogenic and pathogenic microorganisms when assessed by the spot-on-lawn method. Its anitimicrobial activity on sensitive indicator cells disappeared completely by protease XIV treatment. The inhibitory activity of lacticin SA72 remained after treatment for 15 min at $121^{\circ}C$, 문 was stable in a pH range of 2.0 to 9.0 and all organic solvents examined. It demonstrated a typical bactericidal mode of inhibition against Lactobacillus delbrueckii ATCC 4797. The apparent molecular mass of lacticin SA72 was in the region of 3-3.5 kDa, determined by SDS-PAGE.

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Isolation of Biosurfactant-Producing P. aeruginosa Mi-7 and the Biosurfactant Production (Biosurfactant를 생산하는 P. aeruginosa. KK-7의 분리 및 Biosurfactant의 생산)

  • 강상모;김대원;김혜자
    • Microbiology and Biotechnology Letters
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    • v.22 no.1
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    • pp.92-98
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    • 1994
  • The bacteria which secrete surface-active agent and decrease the surface tension of culture broth were isolated from soil samples. Among them, biosurfactant producing strain KK-7 was selected and emulsification was also detected. The KK-7 produced biosurfactant not only lipid but also glucose by using carbon source. Taxonomical characterization tests have demostrated the strain KK-7 to be Pseudomonas aeruginosa. The media composition of the P. aeruginosa KK-7 for the biosurfactant production was 1% glucose, 0.5% tryptone, 0.2% yeast extract, 0.15% potas sium phosphate mono-dibasic, 0.05% MgSO$_{4}$, initial pH 8.5, at 30$\circ $C for 2 days. In this condition, the concentration of biosurfactant was reached CMC 5 in the culture broth. Surface active material was produced maximum at stationary8 phase, but emulsification power was higher at log phase than stationary phase. It was considered that P. aeruginosa KK-7 produced biosurfactant more than one type having defferent properties and each maximum production time was different. The minimun surface tension of biosurfactant in 50 mM Tris buffer (pH8.0) was 28 dyn/cm, and CMC was 1 g/L.

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A Study on The Effect of Ginseng Saponin Fraction on Cell Wall (인삼 사포닌 분획이 세포벽에 미치는 영향)

  • Jo, Yeong-Dong;Kim, Tae-U;Choe, Hae-Gil
    • Journal of Ginseng Research
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    • v.5 no.1
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    • pp.65-72
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    • 1981
  • In this experiment, observations were made on the effects of ginseng saponin, one of the major components of Korean ginseng (Panax ginseng, C. A. Meyer) root, on the membranes of microorganism (E. coli K-12), the concentration of intracelluar and extracellular cycle AMP therein, and uptake of U-14C-glucose. When the E. coli were grown on media containing 0.1% ginseng saponin, the growth was faster than for that of the control by about 30 minutes. The lysis of E. coli grown on the ginseng saponin medium increased to about double that of the control in the stationary phase. And the amount of protein and lipopolysaccharides in the outer cell meberances increased 25% and 80% respectively in comparison with the control. By electron microscope observation, it was shown that the periplasmic region of the E. coli grown on the ginseng saponin medium was widened it was observed that the cellular cyclic AMP content of the E. coli increased significantly to the hightest levels between the late exponential phase and early stationary phase. The total cyclic AMP content of E. coli grown on the ginseng saponin medium decreased about 50% when compared to that of the control.

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Comparative Analysis of Growth-Phase-Dependent Gene Expression in Virulent and Avirulent Streptococcus pneumoniae Using a High-Density DNA Microarray

  • Ko, Kwan Soo;Park, Sulhee;Oh, Won Sup;Suh, Ji-Yoeun;Oh, TaeJeong;Ahn, Sungwhan;Chun, Jongsik;Song, Jae-Hoon
    • Molecules and Cells
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    • v.21 no.1
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    • pp.82-88
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    • 2006
  • The global pattern of growth-dependent gene expression in Streptococcus pneumoniae strains was evaluated using a high-density DNA microarray. Total RNAs obtained from an avirulent S. pneumoniae strain R6 and a virulent strain AMC96-6 were used to compare the expression patterns at seven time points (2.5, 3.5, 4.5, 5.5, 6.0, 6.5, and 8.0 h). The expression profile of strain R6 changed between log and stationary growth (the Log-Stat switch). There were clear differences between the growth-dependent gene expression profiles of the virulent and avirulent pneumococcal strains in 367 of 1,112 genes. Transcripts of genes associated with bacterial competence and capsular polysaccharide formation, as well as clpP and cbpA, were higher in the virulent strain. Our data suggest that late log or early stationary phase may be the most virulent phase of S. pneumoniae.

Analysis and Modeling of Parallel Three-Phase Boost Converters Using Three-Phase Coupled Inductor

  • Lim, Chang-Soon;Lee, Kui-Jun;Kim, Rae-Young;Hyun, Dong-Seok
    • Journal of Electrical Engineering and Technology
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    • v.8 no.5
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    • pp.1086-1095
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    • 2013
  • The main issue of parallel three-phase boost converters is reduction of the low- and high frequency circulating currents. Most present technologies concentrate on low frequency circulating current because the circulating current controller cannot mitigate the high frequency circulating current. In this paper, analytical approach of three-phase coupled inductor applied to parallel system becomes an important objective to effectively reduce the low- and high frequency circulating currents. The characteristics of three-phase coupled inductor based on a structure and voltage equations are mathematically derived. The modified voltage equations are then applied to parallel three-phase boost converters to develop averaged models in stationary coordinates and rotating coordinates. Based on the averaged modeling approach, design of the circulating current controller is presented. Simulation and experimental results demonstrate the effectiveness of the analysis and modeling for the parallel three-phase boost converters using three-phase coupled inductor.

Non-stationary vibration and super-harmonic resonances of nonlinear viscoelastic nano-resonators

  • Ajri, Masoud;Rastgoo, Abbas;Fakhrabadi, Mir Masoud Seyyed
    • Structural Engineering and Mechanics
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    • v.70 no.5
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    • pp.623-637
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    • 2019
  • This paper analyzes the non-stationary vibration and super-harmonic resonances in nonlinear dynamic motion of viscoelastic nano-resonators. For this purpose, a new coupled size-dependent model is developed for a plate-shape nano-resonator made of nonlinear viscoelastic material based on modified coupled stress theory. The virtual work induced by viscous forces obtained in the framework of the Leaderman integral for the size-independent and size-dependent stress tensors. With incorporating the size-dependent potential energy, kinetic energy, and an external excitation force work based on Hamilton's principle, the viscous work equation is balanced. The resulting size-dependent viscoelastically coupled equations are solved using the expansion theory, Galerkin method and the fourth-order Runge-Kutta technique. The Hilbert-Huang transform is performed to examine the effects of the viscoelastic parameter and initial excitation values on the nanosystem free vibration. Furthermore, the secondary resonance due to the super-harmonic motions are examined in the form of frequency response, force response, Poincare map, phase portrait and fast Fourier transforms. The results show that the vibration of viscoelastic nanosystem is non-stationary at higher excitation values unlike the elastic ones. In addition, ignoring the small-size effects shifts the secondary resonance, significantly.

Cloning, Over-expression, and Characterization of YjgA, a Novel ppGpp-binding Protein

  • Gnanasekaran, Gopalsamy;Pan, SangO;Jung, Wontae;Jeong, Kwangjoon;Jeong, Jae-Ho;Rhee, Joon Haeng;Choy, Hyon E.;Jung, Che-Hun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.8
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    • pp.2419-2424
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    • 2013
  • Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

Mutation of the invF Gene Encoding a Salmonella Pathogenicity Island 1 (SPI1) Activator Increases Expression of the SPI2 Gene, sseA (Salmonella Pathogenicity Island 1(SPI1)의 발현조절 유전자 invF의 변이가 SPI2 유전자(sseA)의 발현에 미치는 영향)

  • Han, Ah-Reum;Joe, Min-Ho;Kim, Dong-Ho;Baik, Sang-Ho;Lim, Sang-Yong
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.70-75
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    • 2012
  • In Salmonella enterica, many genes encoded within Salmonella pathogenicity islands (SPI) 1 and 2 are required to cause a range of diseases in a variety of hosts. The SPI1-encoded regulator HilD activates both the SPI1 and 2 genes at different times during growth in Luria-Bertani (LB) media. In this study, the expression levels of hilD during growth in LB were investigated. The data suggest that hilD expression is induced in the early stationary phase and decreases in the late stationary phase, when sseA, an SPI2 gene, is maximally expressed. However, HilD could act as an activator of sseA expression in the late stationary phase despite being present at low levels. SseA expression was investigated in SPI1 regulator mutant strains, hilA, hilD and invF mutants. As expected, hilD mutation decreased sseA expression. However, we found that invF mutation caused a 1.5-fold increase in sseA expression in not only LB but also M9 minimal media, which is thought to resemble an intracellular environment. InvF overexpression restored sseA expression to wild-type levels in an invF mutant but did not cause an additional reduction in sseA expression. These results suggest that SPI1 controls SPI2 expression either positively or negatively.

Comparative Genome-Scale Expression Analysis of Growth Phase-dependent Genes in Wild Type and rpoS Mutant of Escherichia coli

  • Oh, Tae-Jeong;Jung, Il-Lae;Woo, Sook-Kyung;Kim, Myung-Soon;Lee, Sun-Woo;Kim, Keun-Ha;Kim, In-Gyu;An, Sung-Whan
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.258-265
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    • 2004
  • Numerous genes of Escherichia coli have been shown to growth phase-dependent expression throughout growth. The global patterns of growth phase-dependent gene expression of E. coli throughout growth using oligonucleotide microarrays containing a nearly complete set of 4,289 annotated open reading frames. To determine the change of gene expression throughout growth, we compared RNAs taken from timecourses with common reference RNA, which is combined with equal amount of RNA pooled from each time point. The hierarchical clustering of the conditions in accordance with timecourse expression revealed that growth phases were clustered into four classes, consistent with known physiological growth status. We analyzed the differences of expression levels at genome level in both exponential and stationary growth phase cultures. Statistical analysis showed that 213 genes are shown to, growth phase-dependent expression. We also analyzed the expression of 256 known operons and 208 regulatory genes. To assess the global impact of RpoS, we identified 193 genes coregulated with rpoS and their expression levels were examined in the isogenic rpoS mutant. The results revealed that 99 of 193 were novel RpoS-dependent stationary phase-induced genes and the majority of those are functionally unknown. Our data provide that global changes and adjustments of gene expression are coordinately regulated by growth transition in E. coli.

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