• Title/Summary/Keyword: cyanobacterium Synechocystis PCC6803

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Effects of Heavy Metals on Growth and Protein Synthesis in Cyanobacterium synechocystis sp. PCC 6803 (중금속이 Cyanobacterium synechocystis sp.PCC 6803의 성장과 단백질 합성에 미치는 영향)

  • 강경미;장남기
    • Asian Journal of Turfgrass Science
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    • v.10 no.4
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    • pp.315-329
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    • 1996
  • The changes of growth and protein synthesis pattern by aluminum (Al), cadmium (Cd), zinc (Zn) treatments were studied in Cyanobacterium synechocystis sp. PCC 6803. When exposed to Al from 5ppm to 3oppm, synechocystis grows normally. But more than that retard the growth of algae notably. The 0.05ppm Cd additions had no effect on the growth of algae. 0.1, 0.2, and 0.5ppm Cd inhibited growth. Under 1 and 2ppm Cd stress, growth was greatly diminished. Zn had dual effects. The growth of algae in media containing 5ppm was stimulated. As concentration increases more than l5ppm, growth inbition increases. Under 25ppm Zn stress, growth was greatly diminished. According to logistic theory, r and K values of each heavy metal-treated groups were estimated. Correlation analysis of r and K values with metal concentration shows that there is negative correlation between K and concentration in Cd and Zn treatments. Critical concentration which shows lethal or sublethal effect was estimated by t-test of each r and K value. The cells cultured in 10, 20, 30, 40 and 5oppm of Al, 1 and 2ppm of Cd, and 10, 15, 20, 25 and 30ppm of Zn for 4 days was used for protein analysis. Analysis of protein synthesis with SDS-PACE showed alterations of protein synthesis pattern. The synthesis of protein about 220kD increased markedly. In this study, it showed that resistance mechanism against Al, Cd, and Zn is K selection and that metal stress induced the change of protein synthesis in Cyanobacterium synechocystis sp. PCC 6803.Key words:Cyanobacterium synechocystis sp. FCC 6803, Heavy metals, Aluminum, Cadmiutm Zinc, Crowth, Frotein synthesis.

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Putrescine Transport in a Cyanobacterium Synechocystis sp. PCC 6803

  • Raksajit, Wuttinun;Maenpaa, Pirkko;Incharoensakdi, Aran
    • BMB Reports
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    • v.39 no.4
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    • pp.394-399
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    • 2006
  • The transport of putrescine into a moderately salt tolerant cyanobacterium Synechocystis sp. PCC 6803 was characterized by measuring the uptake of radioactively-labeled putrescine. Putrescine transport showed saturation kinetics with an apparent $K_m$ of $92{\pm}10\;{\mu}M$ and $V_{max}$ of $0.33{\pm}0.05\;nmol/min/mg$ protein. The transport of putrescine was pH-dependent with highest activity at pH 7.0. Strong inhibition of putrescine transport was caused by spermine and spermidine whereas only slight inhibition was observed by the addition of various amino acids. These results suggest that the transport system in Synechocystis sp. PCC 6803 is highly specific for polyamines. Putrescine transport is energy-dependent as evidenced by the inhibition by various metabolic inhibitors and ionophores. Slow growth was observed in cells grown under salt stress. Addition of low concentration of putrescine could restore growth almost to the level observed in the absence of salt stress. Upshift of the external osmolality generated by either NaCl or sorbitol caused an increased putrescine transport with an optimum 2-fold increase at 20 mosmol/kg. The stimulation of putrescine transport mediated by osmotic upshift was abolished in chloramphenicol-treated cells, suggesting possible involvement of an inducible transport system.

Cyanobacterium Synechocystis PCC6803 내에서 Bacillus thuringiensis sunsp. morrisoni PG-14 cryIVD 유전자의 발현

  • Lee, Dae-Weon;Park, Hyun-Woo;Jin, Byung-Rae;Chung, Young-Ho;Park, Young-Mok;Kang, Seok-Kwon
    • Microbiology and Biotechnology Letters
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    • v.24 no.2
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    • pp.173-177
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    • 1996
  • Bacillus thuringiensis subsp. morrisoni PG-14 is a gram-positive soil bacterium producing mosquitocidal parasporal inclusions composed of several crystal proteins. Among these crystal protein genes, cryIVD gene is one of major component which has 72 kDa in size. However, these parasporal inclusions sink quickly from the surface of water where mosquito larval feeding occurred. To develope mosquitocidal cyanobacterium, therefore, we constructed the expression vector, pCYASK 5-1 harboring cryIVD gene. The expression vector, pCYASK5-1 was transformed into the cyanobacterium Syne- chocystis PCC6803 reported as a natural mosquito larval food source and the transformants were selected with kanamycin. Expression of IVD gene in transformant was characterized by SDS-polyacrylamide gel electrophoresis (PAGE) and immunoblot analysis. The mosquitocidal activity of a transformant was determined with Culex tritaeniorhynchus. The results showed that, the transformed cyanobacterium is toxic to mosquito larvae and will be expected as a potential agent that is used for mosquito control.

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Influence of Respiration on Photosynthetic Electron Transport in psaB Mutants from Cyanobacterium Synechocystis sp. PCC6803 (Cyanobacterium Synechocystis sp.PCC6803 psaB 돌연변이주의 광합성 전자전달에 미치는 호흡의 영향)

  • 윤병철;장남기
    • Asian Journal of Turfgrass Science
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    • v.11 no.1
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    • pp.59-72
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    • 1997
  • The influence of respiration on photosythetic electron transport were investigated in the Wid type and psaB mutants from Syneehocystis sp. PCC6803. The amount of glucose uptake in the wild type was proportional to the glucose concentration added in wild type and less than that of psaB mutants in the dark. It was suggested that psaB mutants more depend on the glucose than the wild type. It was investigated how the activities of isocitrate dehydrogenase(IDH) and glucose-6-phos-phate dehydrogenase(G6PDH) were changed. The activities of IDH were very low. While, the ac-tivities of G6PDH were much higher than that of IDH. These results agree to the reports that ex-ogenous glucose was dismilated aerobically via Oxidative Pentose Phosphate Pathway in heterotrophic cyanobacteria. PsaB mutants showed high G6PDH activity in the presence of glucose as well as in the dark and high respiratory activities especially in the dark. It was also investigated how photosynthetic electron transport activities were changed. PsaB mutants showed higher photosynthetic electron tranasport activities than wild type in the presence of glucose as well as in the dark. In the results, it was proposed that photosynthetic electron transport between PS I and PS U was complemented by respiratory electron transport through the NADPH generated by Dxidative Pentose Phophate Pathway in psaB mutant from Synechocystis sp. PCC6803. Key words: Photosynthetic electron transport, Respiration, Synechoystis sp. PCC6803, psaB mutant, Glucose uptake, IDH, G6PDH, Respiratory electron transport activity.

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Photokinesis of Cyanobacterium Synechocystis sp. PCC 6803

  • Chung, Young-Ho;Park, Young-Mok;Moon, Yoon-Jung;Lee, Eun-Mi;Choi, Jong-Soon
    • Journal of Photoscience
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    • v.11 no.3
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    • pp.89-94
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    • 2004
  • Motile cyanobacterium Synechocystis sp. PCC 6803 cells show photomovement with respect to the light stimulus. Under lateral irradiation, Synechocystis displays a phototactic gliding movement toward the light source by a twodimensional random biased walk. Under vertical irradiation, Synechocystis decreased the frequency of mean vectorial gliding speed dependent on the applied fluence rate, whereas the deviation distribution width of the speed increased. This strongly suggests the involvement of photokinesis. Evidence for the cyanobacterial photokinesis was discussed in the previous report (Choi et al., 1999. Photochem. Photobiol. 70, 95-102) demonstrating that the gross scalar speed of vertically irradiating cells increased by about 50% compared with that of dark-adapted cells. In the visible wavelength range, Synechocystis cells showed a maximal photokinetic activity at 420 nm and a second maximal activity at 680 nm. The threshold action spectrum for the photokinesis resembles the absorption spectrum of chlorophyll with major differences in the phototaxis action spectrum at 560 nm and 660 nm. We postulate that the cyanobacterial photokinesis is powered by the energy-generating chlorophyll pigments.

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High Throughput Proteomic Approaches for the Dissection of Light Signal Transduction Pathways in Photosynthetic Cyanobacterium Synechocystis sp.PCC 6803

  • Chung Young-Ho;Park Young Mok
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.203-205
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    • 2002
  • Light is an environmental signal that regulates photomovement and main energy source of photosynthesis in the cyanobacterium Synechocystis sp. PCC 6803 (Syn6803). Syn6803 is a popular model system for study of plant functional genomics. In this report, we adopted 2D gel based proteomics study to investigate proteins related with the light absorption and photo-protection in Syn6803. More than 700 proteins were detected on the SDS-gels stained with silver nitrate. Several proteins showing different expression level under various light conditions were identified with MALDI-TOF Mass spectrometry. As a comparison, we also conducted ICAT-based proteome study using WT and cphl (cyanobacterial phytochrome 1) mutant. A cphl deletion led to changes in the expression of proteins involved in translation, photosynthesis including photosystem and CO2 fixation, and cellular regulation. We are currently involved in TAP-tagging method to study protein-protein interactions in search for the molecular component involved in the light signal transduction of Syn6803 photomovement.

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Optimum Conditions for Transformation of Synechocystis sp. PCC 6803

  • Zang, Xiaonan;Liu, Bin;Liu, Shunmei;Arunakumara, K.K.I.U.;Zhang, Xuecheng
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.241-245
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    • 2007
  • This study was conducted to determine the optimal conditions for introduction of exogenous DNA into Synechocystis sp. PCC 6803. Of the three transformation techniques studied, electroporation, ultrasonic transformation and natural transformation, natural transformation showed the highest efficiency. Additionally, this study demonstrated that the higher plasmid concentration and longer homologous recombining fragments resulted in a greater number of transformants. For successful transformation, the lowest concentration of plasmid was $0.02\;{\mu}g/ml$, and the shortest homologous recombining fragment was 0.2 kb. Use of Synechocystis sp. PCC 6803 in the logarithmic growth phase resulted in two-fold higher transformation rate than that of the same organism when cells in the latent phase or the plateau phase were used for transformation. Pretreatment of the host strain, Synechocystis sp. PCC 6803, with EDTA (2 mM) for two days prior to transformation increased the transformation efficiency by 23%. Additionally, incubation of the cells and DNA for 5 h under light conditions increased the transformation efficiency by two orders of magnitude. Moreover, recovery treatment of the cells before they were plated onto antibiotic medium also increased the transformation efficiency.

Multi-component kinetics for the growth of the cyanobacterium Synechocystis sp. PCC6803

  • Kim, Hyun-Woo;Park, Seongjun;Rittmann, Bruce E.
    • Environmental Engineering Research
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    • v.20 no.4
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    • pp.347-355
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    • 2015
  • The growth kinetics of phototrophic microorganisms can be controlled by the light irradiance, the concentration of an inorganic nutrient, or both. A multi-component kinetic model is proposed and tested in novel batch experiments that allow the kinetic parameters for each factor to be estimated independently. For the cyanobacterium Synechocystis sp. PCC6803, the estimated parameters are maximum specific growth rate $({\mu}_{max})=2.8/d$, half-maximum-rate light irradiance $(K_L)=11W/m^2$, half-inhibition-rate light irradiance $(K_{L,I})=39W/m^2$, and half-maximum-rate concentration for inorganic carbon $(K_{S,Ci})=0.5mgC/L$, half-maximum-rate concentration for inorganic nitrogen $(K_{S,Ni})=1.4mgN/L$, and half-maximum-rate concentration for inorganic phosphorus $(K_{S,Pi})=0.06mgP/L$. Compared to other phototrophs having ${\mu}max$ estimates, PCC6803 is a fast-growing r-strategist relying on reaction rate. Its half-maximum-rate and half-inhibition rate values identify the ranges of light irradiance and nutrient concentrations that PCC6803 needs to achieve a high specific growth rate to be a sustainable bioenergy source. To gain the advantages of its high maximum specific growth rate, PCC6803 needs to have moderate light illumination ($7-62W/m^2$ for ${\mu}_{syn}{\geq}1/d$) and relatively high nutrient concentrations: $N_i{\geq}2.3 mgN/L$, $P_i{\geq}0.1mgP/L$, and $C_i{\geq}1.0mgC/L$.

Characterization of Spermidine Transport System in a Cyanobacterium, Synechocystis sp. PCC 6803

  • Raksajit, Wuttinun;Yodsang, Panutda;Maenpaa, Pirkko;Incharoensakdi, Aran
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.447-454
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    • 2009
  • The transport of spermidine into a cyanobacterium, Synechocystis sp. pec 6803, was characterized by measuring the uptake of $^{14}C$-spermidine. Spermidine transport was shown to be saturable with an apparent affinity constant ($K_m$) value of $67{\mu}M$ and a maximal velocity ($V_{max}$) value of 0.45 nmol/min/mg protein. Spermidine uptake was pH-dependent with the pH optimum being 8.0. The competition experiment showed strong inhibition of spermidine uptake by putrescine and spermine, whereas amino acids were hardly inhibitory. The inhibition kinetics of spermidine transport by putrescine and spermine was found to be noncompetitive with $K_i$ values of 292 and $432{\mu}M$, respectively. The inhibition of spermidine transport by various metabolic inhibitors and ionophores suggests that spermidine uptake is energy-dependent. The diminution of cell growth was observed in cells grown at a high concentration of NaCl. Addition of a low concentration of spermidine at 0.5 mM relieved growth inhibition by salt stress. Upshift of the external osmolality generated by either NaCl or sorbitol caused an increased spermidine transport with about 30-40% increase at 10 mosmol/kg upshift.