• Korean Journal of Microbiology
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• v.41 no.1
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• pp.67-73
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• 2005

Photoautotrophic bacteria are promising candidates for the production of poly(3-hydroxybutyrate) (PHB) since they can address the critical problem of substrate costs. In this study, we isolated 25 Tn5-inserted mutants of the Synechocystis sp. PCC 6803 which showed enhanced PHB accumulation compared to the wild-type strain. After 5-days cultivation under nitrogen-limited mixotrophic conditions, the intracellular levels of PHB content in these mutants reached up to $10-30\%$ of dry cell weight (DCW) comparable to $4\%$ of DCW in the wild-type strain. Using the method of inverse PCR, the affected genes of the mutants were mapped on the completely known genome sequence of Synechocystis sp. PCC 6803. As a result, the increased PHB accumulation in 5 mutants were found to be resulted from defects of genes coding for NADH-ubiquinone oxidoreductase, O-succinylbenzoic-CoA ligase, photosystem II PsbT protein or histidine kinase, which are involved in photosystem in thylakoid inner membrane of the cell. The values of $NAD(P)H/NAD(P)^+$ ratio in the cells of these mutants were much higher than that of the wild-type strain as measured by using pulse-amplitude modulated fluorometer, suggesting that PHB synthesis could be enhanced by increasing the level of cellular NAD(P)H which is a limiting substrate for NADPH-dependent acetoacetyl-CoA reductase. From these results, it is likely that NAD(P)H would be a limiting factor for PHB synthesis in Synechocystis sp. PCC 6803.

• Proceedings of the Zoological Society Korea Conference
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• 1998.10b
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• pp.304.2-304
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• 1998

• The Korea Genome Conference
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• 2003.09a
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• pp.40.1-40.1
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• 2003

• Korean Journal of Ecology and Environment
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• v.42 no.1
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• pp.67-74
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• 2009

To study the influence of the rotifer Brachionus calyciflorus on harmful algal bloom suppression, we focused on assessing the rotifer's abilities using several prey species : Microcystis aeruginosa, Synechocystis sp., Chlorella vulgaris and Coelastrum sp. of the warm-weather species and the cold-weather centric diatom Stephanodiscus hantzchii. Grazing effects and growth rates of rotifers B. calyciflorus were 94.5% and $1.29d^{-1}$, respectively, for Synechocystis sp., 87.4% and $0.60d^{-1}$, respectively, for M. aeruginosa, 95.2% and $0.65d^{-1}$, respectively, for C. $vulgaris^{TM}$, 78.6% and $0.45d^{-1}$, respectively, for C. vulgaris UTEX., 86.5% and $0.99d^{-1}$, respectively, for Coelastrum sp., and 82.6% and $0.40d^{-1}$, respectively, for S. hantzchii. Of these, although the growth of Synechocystis and Coelastrum was effectively suppressed by rotifer grazing, efficient suppression effects on Stephanodiscus blooms were unexpected. The present study revealed that reproduction of B. calyciflorus was greatly influenced by its food types in the initial stages and the efficiencies of bio-agents as sole food sources vary depending on the target algae and the agent.

• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.407-414
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• 2024

Phosphorus is an essential but non-renewable nutrient resource critical for agriculture. Luxury phosphorus uptake allows microalgae to synthesize polyphosphate and accumulate phosphorus, but, depending on the strain of algae, polyphosphate may be degraded within 4 hours of accumulation. We studied the recovery of phosphorus from wastewater through luxury uptake by an engineered strain of Synechocystis sp. with inhibited polyphosphate degradation and the effect of this engineered Synechocystis biomass on lettuce growth. First, a strain (∆phoU) lacking the phoU gene, which encodes a negative regulator of environmental phosphate concentrations, was generated to inhibit polyphosphate degradation in cells. Polyphosphate concentrations in the phoU knock-out strain were maintained for 24 h and then decreased slowly. In contrast, polyphosphate concentrations in the wild-type strain increased up to 4 h and then decreased rapidly. In addition, polyphosphate concentration in the phoU knockout strain cultured in semi-permeable membrane bioreactors with artificial wastewater medium was 2.5 times higher than that in the wild type and decreased to only 16% after 48 h. The biomass of lettuce treated with the phoU knockout strain (0.157 mg P/m²) was 38% higher than that of the lettuce treated with the control group. These results indicate that treating lettuce with this microalgal biomass can be beneficial to crop growth. These results suggest that the use of polyphosphate-accumulating microalgae as biofertilizers may alleviate the effects of a diminishing phosphorous supply. These findings can be used as a basis for additional genetic engineering to increase intracellular polyphosphate levels.

• 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.

• Proceedings of the Korean Society of Potoscience Conference
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• 2002.06a
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• pp.38-38
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• 2002

• Proceedings of the Korean Society of Potoscience Conference
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• 2002.06a
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• pp.47-47
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• 2002

• Proceedings of the Korean Society of Potoscience Conference
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• 2005.06a
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• pp.111-111
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• 2005

• Journal of Microbiology and Biotechnology
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• v.26 no.5
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• pp.959-966
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• 2016

Chlorophyll synthase (ChlG) and bacteriochlorophyll synthase (BchG) have a high degree of substrate specificity. The BchG mutant of Rhodobacter sphaeroides, BG1 strain, is photosynthetically incompetent. When BG1 harboring chlG of Synechocystis sp. PCC 6803 was cultured photoheterotrophically, colonies arose at a frequency of approximately 10^-8. All the suppressor mutants were determined to have the same mutational change, ChlG_I44F. The mutated enzyme ChlG_I44F showed BchG activity. Remarkably, BchG_F28I, which has the substitution of F at the corresponding 28^th residue to I, showed ChlG activity. The K_m values of ChlG_I44F and BchG_F28I for their original substrates, chlorophyllide (Chlide) a and bacteriochlorophyllide (Bchlide) a, respectively, were not affected by the mutations, but the K_m values of ChlG_I44F and BchG_F28I for the new substrates Bchlide a and Chlide a, respectively, were more than 10-fold larger than those for their original substrates, suggesting the lower affinities for new substrates. Taken together, I44 and F28 are important for the substrate specificities of ChlG and BchG, respectively. The BchG activity of ChlG_I44F and the ChlG activity of BchG_F28I further suggest that ChlG and BchG are evolutionarily related enzymes.