• 제목/요약/키워드: 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
    • 한국동물위생학회지
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    • 제30권3호
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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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    • 제10권4호
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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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Biosurfactant를 생산하는 P. aeruginosa. KK-7의 분리 및 Biosurfactant의 생산 (Isolation of Biosurfactant-Producing P. aeruginosa Mi-7 and the Biosurfactant Production)

  • 강상모;김대원;김혜자
    • 한국미생물·생명공학회지
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    • 제22권1호
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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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인삼 사포닌 분획이 세포벽에 미치는 영향 (A Study on The Effect of Ginseng Saponin Fraction on Cell Wall)

  • 조영동;김태우;최해길
    • Journal of Ginseng Research
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    • 제5권1호
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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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    • 제21권1호
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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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    • 제8권5호
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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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    • 제70권5호
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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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    • 제34권8호
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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.

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

  • 한아름;조민호;김동호;백상호;임상용
    • 한국미생물·생명공학회지
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    • 제40권1호
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    • pp.70-75
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    • 2012
  • 살모넬라(Salmonella)의 염색체에 존재하는 병원성 유전자의 집합체인 Salmonella pathogenicity island(SPI)1 과 2는 살모넬라가 유발하는 다양한 질병에 중요한 역할을 한다. SPI1의 발현을 유도하는 HilD는 Luria-Bertani(LB) 배지 조건에서 SPI2의 발현 활성인자로 작용하는 것으로 알려져 있으나 LB 배지 내에서 hilD 유전자의 발현 양상은 아직까지 연구되지 않았다. 본 연구에서는 LB 배지에 살모넬라를 배양하면서 hilD 유전자의 발현과 단백질 양을 조사하였으며 SPI2 유전자인 sseA의 발현과 비교하였다. hilD의 발현은 대수 증식기 경과 후 정지기(stationary phase)로 전환되는 시기에 비약적으로 증가하였으나 sseA의 발현은 정지기 후반부에 최대로 증가하였다. 즉, 후반 정지기에서 HilD 단백질은 낮은 수준으로 존재함에도 불구하고 SPI2의 발현을 유도한다는 것을 알 수 있었다. SPI1의 다른 발현 조절인자인 hilA와 invF의 변이체에서 sseA의 발현을 살펴본 결과 invF의 변이는 hilD와는 다르게 배지 조건에 상관없이 오히려 sseA의 발현을 증가시켰다. 또한, InvF의 과발현은 sseA 발현을 정상 수준으로 복원시켰지만 추가적인 감소는 일으키지 않는다는 것을 알 수 있었다. SPI1은 HilD를 이용하여 SPI2의 발현을 유도하지만 반대로 InvF를 이용하여 발현을 억제하기도 하는 이중적인 조절 기전을 가지고 있는 것으로 판단된다.

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
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2004년도 Annual Meeting BioExibition International Symposium
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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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