• ALGAE
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• 제18권2호
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• pp.121-127
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• 2003

The pyrenoid ultrastructure and distribution of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and Rubisco activase in the chloroplasts of Chlamydomonas reinhardtii was studied using the immunogold localization technology with electron microscopy. There were several tubular thylakoids invading in the pyrenoid matrix to form several spokewise channels. The connections between pyrenoid matrix and stroma of chloroplast were the partial of channels. The starch sheath surrounding the pyrenoid was separated into several parts by the connections in transection. Some thylakoids were packed together near the connections in one side of the pyrenoid. Those special structures might be used to transport substance between pyrenoid and stroma of chloroplasts. With the antibody raised against the large subunits of Rubsico from C. protothecoides, the result of the gold immunolocalization of Rubisco in Chlamydomonas reinhardtii showed most of the gold particles heavily labeled the pyrenoid matrix, as well as the starch sheath matrix, and very few in the stroma of chloroplasts. The gold particle density was 880.00 $\pm$ 164.32, 190.00 $\pm$ 152.39 and 9.60 $\pm$ 5.37 ${\mu}m^{-2}$ in pyrenoid matrix, starch sheath and stroma region of chloroplast respectively (background: 5.67 $\pm$ 1.53 ${\mu}m^{-2}$). 99.59% of the total Rubiscos was calculated to be concentrated in the pyrenoid matrix and starch sheath by spatial densities. The gold immunolocalization of Rubisco activase also showed that Rubisco activase was mainly concentrated in the periphery of the pyrenoid and the starch sheath (the density was as high as 229.69 $\pm$ 96.96 ${\mu}m^{-2}$). There were very few gold particles located in the stroma of chloroplasts. These results indicated that pyrenoid surface and starch sheath was the site for Rubisco activation and $CO_2$ fixation, which supported the suggestion that pyrenoids perform photosynthesis function.

• 한국수소및신에너지학회논문집
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• 제16권1호
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• pp.17-24
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• 2005

We experimented on growth in light and production of hydrogen and organic matters in dark fermentation by using C. reinhardtii. In the light, growth rate of C. reinhardtii following $CO_2$ fixation was proportional to consumption rate of nitrogen source. And the starch in cell was accumulated more when the period of culture was lengthened more. But the accumulation rate of starch in cell was decreased when the growth rate of cell become dull. In the dark fermentation, the production volume and production rate of hydrogen were the highest value in the mid exponential state among other states. The utilization efficiency of substrate was better in the early exponential state than other states. In production of organic matters, acetic acid didn't change remarkably and ethanol showed the highest value in early exponential state.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1150-1156
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• 2017

Understanding phosphorus metabolism in photosynthetic organisms is important as it is closely associated with enhanced crop productivity and pollution management for natural ecosystems (e.g., algal blooming). Accordingly, we exploited highly time-resolved metabolic responses to different levels of phosphate deprivation in Chlamydomonas reinhardtii, a photosynthetic model organism. We conducted non-targeted primary metabolite profiling using gas-chromatography time-of-flight mass spectrometric analysis. Primarily, we systematically identified main contributors to degree-wise responses corresponding to the levels of phosphate deprivation. Additionally, we systematically characterized the metabolite sets specific to different phosphate conditions and their interactions with culture time. Among them were various types of fatty acids that were most dynamically modulated by the phosphate availability and culture time in addition to phosphorylated compounds.

• Journal of Microbiology and Biotechnology
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• 제28권12호
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• pp.2121-2132
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• 2018

Abnormal melanin synthesis results in several hyperpigmentary disorders such as freckles, melanoderma, age spots, and other related conditions. In this study, we investigated the anti-melanogenic effects of an extract from the microalgae Chlamydomonas reinhardtii (CE) and potential mechanisms responsible for its inhibitory effect in B16F10, normal human epidermal melanocyte cells, and human skin-equivalent models. The CE extract showed significant dose-dependent inhibitory effects on ${\alpha}$-melanocyte-stimulating, hormone-induced melanin synthesis in cells. Additionally, the CE extract exhibited suppressive effects on the mRNA and protein expression of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein-1, and tyrosinase-related protein-2. The CE extract also inhibited the phosphorylation of protein kinase A and extracellular signal-related kinase, which function as upstream regulators of melanogenesis. Using a three-dimensional, reconstructed pigmented epidermis model, the CE-mediated, anti-pigmentation effects were confirmed by Fontana-Masson staining and melanin content assays. Taken together, CE extract can be used as an anti-pigmentation agent.

• 한국물환경학회지
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• 제21권4호
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• pp.332-336
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• 2005

The average specific growth rate of C. reinhardtii (${\mu}$) was decreased with increase in biocide concentrations. The toxicity of biocides toward was as follows (in descending order of toxicities): herbicide 〉 pesticide 〉fungicide. $EC_{50}$ in each biocide was 0.0017 mg/L, 1.06 mg/L and 13.3 mg/L for herbicide, pesticide and fungicide respectively. When herbicide and pesticide were mixed, $EC_{50}$ was decreased by $2.7{\times}10^{-7}mg/L$. $EC_{50}$ in effective components of each biocide was 5.26 mg/L, 9.37 mg/L and 20.58 mg/L for herbicide, pesticide and fungicide respectively. Mixed main components of herbicide and pesticide caused to decrease by 3.10 mg/L.

• Journal of Plant Biology
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• 제32권4호
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• pp.331-341
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• 1989

DNA methyltransferase was purified 282.6-fold from Chlamydomonas reinhardtii 21gr (mt+) gametic cell to examine the effect of polyamine on the enzyme acctivity. Polyacrylamide gel electrophoresis(PAGE) revealed at least three bands(1 major band, 2 minor bands). Among these, the major band represents DNA methyltransferase. Polyacrylamide gel electrophoresis in the presence of 0.1% sodium dodecylsulfate(SDS-PAGE) revealed a major band with M.W. 60,000. DNA methyltransferase activity was inhibited more effectively by spermine than by spermidine, and the inhibition by putrescine was smaller than spermine and spermidine. DNA methyltransferase activity was inhibited by 40% and 53% at 5mM and 20mM spermine, respectively. In the case of spermidine, the inhibition was 35% at 10mM and 44% at 20mM. However, the inhibition by putrescine appeared only above 5mM and reached about 25% at 20mM.

• Journal of Microbiology and Biotechnology
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• 제23권7호
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• pp.923-931
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• 2013

Rapamycin, known as an inhibitor of Target of Rapamycin (TOR), is an immunosuppressant drug used to prevent rejection in organ transplantation. Despite the close association of the TOR signaling cascade with various scopes of metabolism, it has not yet been thoroughly investigated at the metabolome level. In our current study, we applied mass spectrometric analysis for profiling primary metabolism in order to capture the responsive dynamics of the Chlamydomonas metabolome to the inhibition of TOR by rapamycin. Accordingly, we identified the impact of the rapamycin treatment at the level of metabolomic phenotypes that were clearly distinguished by multivariate statistical analysis. Pathway analysis pinpointed that inactivation of the TCA cycle was accompanied by the inhibition of cellular growth. Relative to the constant suppression of the TCA cycle, most amino acids were significantly increased in a time-dependent manner by longer exposure to rapamycin treatment, after an initial down-regulation at the early stage of exposure. Finally, we explored the isolation of the responsive metabolic factors into the rapamycin treatment and the culture duration, respectively.

• Journal of Microbiology and Biotechnology
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• 제30권11호
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• pp.1777-1784
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• 2020

To increase the availability of microalgae as producers of valuable compounds, it is necessary to develop novel systems for gene expression regulation. Among the diverse expression systems available in microalgae, none are designed to induce expression by low temperature. In this study, we explored a cold-inducible system using the antifreeze protein (AFP) promoter from a polar diatom, Chaetoceros neogracile. A vector containing the CnAFP promoter (pCnAFP) was generated to regulate nuclear gene expression, and reporter genes (Gaussia luciferase (GLuc) and mVenus fluorescent protein (mVenus)) were successfully expressed in the model microalga, Chlamydomonas reinhardtii. In particular, under the control of pCnAFP, the expression of these genes was increased at low temperature, unlike pAR1, a promoter that is widely used for gene expression in C. reinhardtii. Promoter truncation assays showed that cold inducibility was still present even when pCnAFP was shortened to 600 bp, indicating the presence of a low-temperature response element between -600 and -477 bp. Our results show the availability of new heterologous gene expression systems with cold-inducible promoters and the possibility to find novel low-temperature response factors in microalgae. Through further improvement, this cold-inducible promoter could be used to develop more efficient expression tools.

• Molecules and Cells
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• 제43권1호
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• pp.48-57
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• 2020

The microalga Chlamydomonas reinhardtii accumulates triacylglycerols (TAGs) in lipid droplets under stress conditions, such as nitrogen starvation. TAG biosynthesis occurs mainly at the endoplasmic reticulum (ER) and requires fatty acid (FA) substrates supplied from chloroplasts. How FAs are transferred from chloroplast to ER in microalgae was unknown. We previously reported that an Arabidopsis thaliana ATP-binding cassette (ABC) transporter, AtABCA9, facilitates FA transport at the ER during seed development. Here we identified a gene homologous to AtABCA9 in the C. reinhardtii genome, which we named CrABCA2. Under nitrogen deprivation conditions, CrABCA2 expression was upregulated, and the CrABCA2 protein level also increased. CrABCA2 knockdown lines accumulated less TAGs and CrABCA2 overexpression lines accumulated more TAGs than their untransformed parental lines. Transmission electron microscopy showed that CrABCA2 was localized in swollen ER. These results suggest that CrABCA2 transports substrates for TAG biosynthesis to the ER during nitrogen starvation. Our study provides a potential tool for increasing lipid production in microalgae.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1998년도 한국생물과학협회 학술발표대회
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• pp.256.1-256
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• 1998

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