• Journal of Plant Biology
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• 제35권4호
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• pp.415-423
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• 1992

Chlamydomonas reinhardtii 21 gr(mt+) strain의 배우체로부터 두 종류의 DNA methylase를 부분 분리하여 몇가지 기질 DNA에 대한 효소 활성을 측정하였다. DNA methylase I과 II는 동일한 pH와 ionic strength에서 서로 상이한 물리적인 성질과 서로 다른 분자량을 가지며 DNA methylase I과 II는 모두가 DNA 염기 중 adenine보다는 cytosine에 methylation을 수행하는 것으로 생각된다. 합성 DNA를 사용한 실험에서 DNA methylase I과는 달리 DNA methylase II는 poly(dA-dC)·poly(dG-dT)에서 보다 poly(dG-dC)·poly(dG-dC)의 oligonucleotide에서 더 높은 효소활성을 나타내었다. Chlamydomonas reinhardtii에서 추출한 엽록체 DNA를 기질로 사용하였을 때 DNA methylase I과 II 모두가 배우체기 보다는 영양생장기의 엽록체 DNA에 더 높은 활성을 나타내었다.

• 미생물학회지
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• 제30권3호
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• pp.225-231
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• 1992

생물체내의 ascorbic acid :dehydroascorbic acid system 을 조절하는데 관여하는 것으로 알려진 ascorbate 산화효소의 활성을 단세포 녹조류인 Chlamydomonas reinhardtii 에서 확인함으로써 이 효소의 존재와 특성을 고찰하였다. 조효소는 native gel 전기 영동에 의한 활성 염색과 분광광도계에 의한 ascorbic acid .의 흡광도 감소 측정에서 뚜렷한 활성을 나타내었다. 황산암모늄 침전, hydroxyapatite 흡착 크로마토그래피, 그리고 Sephadex G-150 gel 여과 크로마토그래피 분리한 결과 26.1 units/mg /의 specific activity 를 나타내었다. 분리된 ascorbate 산화효소는 $55^{\circ}C$, pH 4.6 에서 가장 활성도가 높았다. Native gel 전기 영동에서 88,000 dalton 갸량의 분자량을 가지고 있었고 소단위체의 분자량을 55,000 이었다. 또한 Western bloting technique 을 이용하여 C. reinhardtii 의 ascorbate oxidase를 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제15권6호
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• pp.1159-1163
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• 2005

The production of hydrogen by Chlamydomonas reinhardtii UTEX 90, a marine green alga, was performed under dark fermentation. The effects of initial nitrogen and phosphorus concentration on the cell growth and the production of hydrogen and organic substances were investigated. In the growth stage, the maximum dry cell weight (DCW) was 3 g/l when the initial ammonium concentration was 15 mM. In the dark fermentation, the maximum hydrogen production was $3.5\;{\mu}mol/\;mg$ DCW when the initial nitrogen concentration was 7.5 mM. The nitrogen concentration had a greater effect on organic compound and hydrogen production than the phosphorus concentration during the dark fermentation. An investigation of the duration of dark fermentation showed that, at least until three days, dark fermentation should be prolonged for maximum hydrogen production.

• Journal of Plant Biology
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• 제34권4호
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• pp.267-273
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• 1991

Polyamine levels in the male and female cells as well as DNA methyltransferase activity in the female cells during gametogenesis of Chlamydomonas reinhardtii indicated that both spermidine and spermine levels were decreased while DNA methyltransferase activity was markedly increased about 12 hours after the onset of gametogenesis. In vitro, putrescine and spermine at 1 mM inhibited methylation of chloroplasts DNA isolated from vegetative female cells by 35% and 65%, respectively. Spermine was found to be more inhibitory than putrescine at all concentrations tested. The pattern of the inhibition by polyamines appeared different from that caused by cations. The results obtained in this work suggest that the polyamine inhibition of DNA methylation is due to an action of polyamines on the enzyme involved instead of on the DNA itself.

• 한국수소및신에너지학회논문집
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• 제15권4호
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• pp.309-316
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• 2004

Chlamydomonas reinhardtii UTEX 90 was cultivated with continuous supply of 2% $CO_2$ using TAP media at $25^\circ{C}$ and produced biomass 1.18 g of dry cell weight/L for 4 days. C. reinhardtii algal biomass(CAB) was concentrated to 20 times by volume and converted into hydrogen and organic acids by anaerobic fermentation using Clostridium butyricum. Organic acids in the fermentate of CAB were consecutively used to produce hydrogen by Rhodobacter sphaeroides KD 131 under the light condition. Approximately 52% of starch in the concentrated CAB which had 4-5.8, 24-26 and 6-7 g/L of starch, protein and fat, respectively was degraded by Cl. butyricum at $37^\circ{C}$. During this process, hydrogen and some organic acids, such as formate, acetate, propionate, and butyrate, respectively were produced. Further conversion of the organic acids in anaerobic fermentate of CAB by Rb. sphaeroides KD131 produced hydrogen from the anaerobic fermentate under the illumination of 8 klux using halogen lamp at $30^\circ{C}$. The result showed that hydrogen was evolved by the anaerobic conversion using Cl. butyricum and then by the photosynthetic fermentation using Rb. sphaeroides KD131. It indicated that the two-step conversion process produced the maximum amount of hydrogen from algal biomass which contained carbohydrate, protein, and fat via organic acids.

• Journal of Microbiology and Biotechnology
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• 제26권12호
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• pp.2066-2075
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• 2016

In this study, an enhanced lipid-producing mutant strain of the microalga Chlamydomonas reinhardtii was developed by gamma irradiation. To induce the mutation, C. reinhardtii was gamma irradiated at a dose of 400 Gy. After irradiation, the surviving cells were stained with Nile red. The mutant (Cr-4013) accumulating 20% more lipid than the wild type was selected. Thin-layer chromatography revealed the triglyceride and free fatty acid contents to be markedly increased in Cr-4013. The major fatty acids identified were palmitic acid, oleic acid, linoleic acid, and linolenic acid. Random amplified polymeric DNA analysis showed partial genetic modifications in Cr-4013. To ascertain the changes of protein expression in the mutant strain, two-dimensional electrophoresis was conducted. These results showed that gamma radiation could be used for the development of efficient microalgal strains for lipid production.

• Journal of Microbiology and Biotechnology
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• 제23권3호
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• pp.375-381
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• 2013

A freshwater microalga, Chlamydomonas reinhardtii KNUA021, was characterized for its potential as a biochemical feedstock. Its optimal growth was observed when the culture was incubated at $25^{\circ}C$ and pH 9.4. However, the isolate was capable of survival and growth under a variety of temperatures (10-$30^{\circ}C$) and pH (pH 4.0-12.0) conditions. The total lipid content of the isolate was 21.7% of dry weight and it was found that a high-value fatty alcohol, hexadecenol ($C_{20}H_{40}O$), was autotrophically produced by strain KNUA021. In addition, a nutritionally important $C_{18:3}{\omega}3$ (${\alpha}$-linolenic acid, ALA) was also identified in this photosynthetic microorganism as one of the major fatty acids. Hence, C. reinhardtii KNUA021 appears to show promise for use in the production of microalgae-based biochemicals.

• Journal of Microbiology and Biotechnology
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• 제33권3호
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• pp.310-318
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• 2023

Microalgae are attracting much attention as promising, eco-friendly producers of bioenergy due to their fast growth, absorption of carbon dioxide from the atmosphere, and production capacity in wastewater and salt water. However, microalgae can only accumulate large quantities of lipid in abiotic stress, which reduces productivity by decreasing cell growth. In this study, the strategy was investigated to increase cell viability and lipid production by overexpressing S-adenosylmethionine (SAM) synthetase (SAMS) in the microalga Chlamydomonas reinhardtii. SAM is a substance that plays an important role in various intracellular biochemical reactions, such as cell proliferation and stress response, and the overexpression of SAMS could allow cells to ithstand the abiotic stress and increase productivity. Compared to wild-type C. reinhardtii, recombinant cells overexpressing SAMS grew 1.56-fold faster and produced 1.51-fold more lipids in a nitrogen-depleted medium. Furthermore, under saline-stress conditions, the survival rate and lipid accumulation were 1.56 and 2.04 times higher in the SAMS-overexpressing strain, respectively. These results suggest that the overexpression of SAMS in recombinant C. reinhardtii has high potential in the industrial-scale production of biofuels and various other high-value-added materials.

• ALGAE
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• 제34권1호
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• pp.35-43
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• 2019

Genetic study of haploid organisms offers the advantage that mutant phenotypes are directly displayed, but has the disadvantage that strains carrying lethal mutations are not readily maintained. We describe an approach for generating and performing genetic analysis of diploid strains of Chlamydomonas reinhardtii, which is normally haploid. First protocol utilizes self-mating diploid strains that will facilitate the genetic analysis of recessive lethal mutations by offering a convenient way to produce homozygous diploids in a single mating. Second protocol is designed to reduce the chance of contamination and the accumulation of spontaneous mutations for long-term storage of mutant strains. Third protocol for inducing the meiotic program is also included to produce haploid mutant strains following tetraploid genetic analysis. We discuss implication of self-fertile strains for the future of Chlamydomonas research.

• Journal of Plant Biology
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• 제37권1호
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• pp.101-109
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• 1994

Chlamydomonas reinhardtii 배우체 유도기 동안에 polyamine 생합성 억제제인 MGBG[methylglyoxal bis-(guanylhydrazone); SAMDC 억제제] 1mM 처리구와 정상 대조구에서의 polyamine 함량 변화와 Chlamydomonas의 ct-DNA methylation과의 상호 연관성을 조사한 결과, 전반적으로 137C(＋)와 137C(-)의 정상 대조구에서 polyamine 수준은 배우체 유도기 전반에 걸쳐서 감소추세로 나타났으나, 특히 137C(＋)가 137C(-)보다 polyamine 함량이 많았으며 동시에 감소량도 컸다. 1mM MGBG 처리구에서는 137C(＋)의 spermidine 함량이 정상 대조구보다 증가하였다. In vitro에서 ctDNA methylation에 대한 MGBG의 영향은 정상 대조구에서보다도 1mM MGBG 처리구에서 약 20-30%의 ctDNA methylation 감소를 나타내었다. 또한, MGBG는 in vitro에서 DNA methylase에 대하여 억제효과를 나타내어TEk.