• 한국해양바이오학회지
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• 제8권1호
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• pp.18-23
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• 2016

It is important to design photobioreactor by cheap material for economical microalgal biomass production. In this study, two types of marine photobioreactors (MPBR), made by either polyvinyl chloride (MPBR-PVC) or high density poly ethylene (MPBR-HDPE), are used and performance of these were compared. Tetraselmis sp. KCTC 12236BP is a green marine alga that isolated from Ganghwa Island, Korea, and the strain was used for marine cultivations using MPBR-PVC and MPBR-HDPE. The cultivations were performed three times in the spring season of 2012 using MPBR-PVC and of 2013 using MPBR-HDPE in the coastal area of Young Heung Island. As the results, MPBR-PVC shows higher biomass productivities than MPBR-HDPE, due to its high light transmittance. In the cultivations using MPBR-PVC, the average sea water temperature was $11.5^{\circ}C$ during the first experiment and $16.5^{\circ}C$ during the second and third experiments. Average light intensities during three times for experiments were 407.5, 268.1 and $273.0{\mu}{\cdot}E{\cdot}m^{-2}{\cdot}s^{-1}$, respectively. The maximum fresh cell weight and average biomass productivity were $1.2g{\cdot}L^{-1}$ and $0.12g{\cdot}L^{-1}{\cdot}day^{-1}$. These results showed that Tetraselmis sp. KCTC12236BP were adapted well with the environmental conditions from ocean, and grow in the MPBR-PVC and MPBR-HDPE.

• 미생물학회지
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• 제50권4호
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• pp.313-318
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• 2014

최근 지구온난화문제와 화석연료 자원의 부족문제로 인해 지속적이고 친환경적인 미세조류 기반 바이오디젤이 주목받고 있다. 본 연구에서는 돈분 액체비료와 농업용 복합비료가 바이오 디젤 생산을 위한 미세조류 영양원으로 활용될 가능성을 확인해 보았다. 혼합 미세조류 CSS (Chlorella spp., Scenedesmus spp., Stigeoclonium spp.) 배양을 위한 영양원은 돈분 액체비료, 돈분 액체비료와 농업용 복합비료의 혼합, 농업용 복합비료이며, 28일간 Small Scale Raceway Pond (SSRP)에서 옥외배양하였다. 그 결과 혼합 실험구의 바이오매스와 지질 생산성은 각각 0.8g/L, 5.8 mg/L/day로 가장 우수한 바이오매스량과 지질 생산성을 보여주었다. 이 연구를 통해 농업용 복합비료와 돈분 액체비료의 혼합액이 미세조류에 안정적인 영양 공급을 하는 영양원으로 활용될 가능성을 확인하였다. 또한, 미세조류 배양단가와 바이오디젤 생산단가를 낮출 가능성도 탐구하였다.

• 생명과학회지
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• 제32권3호
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• pp.202-209
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• 2022

반수체 단세포 진핵생물인 녹조류 Chlamydomonas reinhardtii는 미세조류와 유전자 변형을 이용하여 고부가가치 물질을 생산하는 세포 공장으로 연구자와 산업계에서 오랫동안 사용되어 왔다. 현재 대부분의 기초 및 산업 연구에 사용되는 외래종인 C. reinhardtii를 대체하기 위하여 충청과 전라지역 12개의 담수에서 채취한 시료에서 Chlamydomonas로 추정되는 미세조류 K01을 분리하였다. 분리균주 K01의 형태학적 분석과 18S rDNA 유전자 서열(NCBI 수탁번호 KC166137)의 계통발생학적 분석을 통하여 분리균주 K01는 Chlamydomonas reinhardtii로 동정 되었다. 분리균주 C. reinhardtii K01의 성장 및 지질 함량은 TAP 배지에서 3개의 야생 균주 및 4개의 돌연변이 균주와 비교하였다. 이들 균주 중 C. reinhardtii K01 균주가 배양 3일째 1.74×10⁷ cells/ml개의 세포수가 관찰되었다. 이는 비교 균주 중 가장 높은 세포 수(1.20×10⁷ cells/ml)를 나타내어 UTEX2244와 비교했을 때 1.5배 빠른 성장속도를 보였다. 분리균주 C. reinhardtii K01의 지질함량(20.67%)은 야생형 균주의 지질함량과 유사하였다. 분리균주 Chlamydomonas reinhardtii K01의 지방산 성분은 7종의 분양 균주에서 확인된 지방산 중 oleate (C18:1)가 검출되지 않았다. 분리균주 C. reinhardtii K01은 nitrate (NaNO₃)를 질소원으로 포함하는 BG11배지에서 배양 6일 동안 0.87 g/l까지 세포밀도가 증가하였으나, 분양균주인 7종의 Chlamydomonas의 경우 모든 균주에서 세포증식이 거의 일어나지 않았다.

• 수산해양기술연구
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• 제50권4호
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• pp.447-454
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• 2014

The marine microalgae Chaetoceros calcitrans was cultured under a fluorescent lamp (CON) and light-emitting diodes (LEDs) of various wavelengths (blue, LB; red, LR; green, LG; white, LW); changes in growth, fucoxanthin, chlorophyll-a, amino acid and fatty acid profiles were investigated. LR-exposed cultures exhibited the highest specific growth rate (SGR) (0.34), whereas LG-exposed cultures showed the lowest SGR (0.26). After cultivation for 10 days, the maximum dry cell weight (g/L) of LR-exposed cultures was significantly higher than that of those exposed to other light conditions (LR${\geq_-}$CON>LB${\geq_-}$LW${\geq_-}$LG). Eicosapentaenoic acid (EPA) levels were significantly higher in CON-exposed cultures compared to those exposed to LW (P<0.05), with no marked difference compared to those exposed to LB, LR and LG (P>0.05). The fucoxanthin content was highest in LB-exposed cultures ($6.3{\mu}g/mL$), whereas LW showed the lowest ($3.6{\mu}g/mL$; P<0.05). Chlorophyll-a content was highest in cultures exposed to LB compared to other light sources. These results suggest consistent differences in growth and biochemical composition after exposure to light of different wavelengths.

• Preventive Nutrition and Food Science
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• 제14권1호
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• pp.21-28
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• 2009

In this study, we focused on natural water-soluble antioxidants from Tetraselmis suecica (T. suecica) and Chlorella ellipsoidea (C. ellipsoidea). They were prepared by enzymatic digestion using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Alcalase, Flavourzyme, Neutrase, and Kojizyme), and the potential antioxidant activity of each was assessed. Most enzymatic digests from T. suecica had a higher radical scavenging activity than those from C. ellipsoidea. Among the enzymatic digests, Kojizyme digest from T. suecica exhibited the highest effect on DPPH radical scavenging. Viscozyme (30.2%) and Neutrase (34.6%) digests from T. suecica exhibited higher hydroxyl radical scavenging activity. Kojizyme digest from T. suecica (81.5%) had strong alkyl radical scavenging activity. Neutrase (61.9%) and Kojizyme (61.5%) digest from T. suecica possessed the highest effects on hydrogen peroxide scavenging. Among the tested samples, Neutrase (TN) and Kojizyme (TK) digests from T. suecica showed the highest antioxidant activity (DPPH, alkyl radical, hydrogen peroxide). Therefore, TN and TK digests were selected for use in the further experiments. Those digests showed enhanced cell viability against $H_2O_2$-induced oxidative damage, and relatively good hydrogen peroxide scavenging activity in an African green monkey kidney (Vero) cell line. These results suggested that an enzymatic digestion will be an effective way for the production of a potential water-soluble antioxidant from a microalgae, T. suecica.

• ALGAE
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• 제27권4호
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• pp.295-301
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• 2012

Carbon concentrating mechanisms play a vital role in photosynthesis in microalgae and cyanobacteria especially in the proper functioning of Rubisco and assimilation of carbon via the Calvin cycle. This study evaluates the role of carbon dioxide on carbon concentrating mechanism (CCM) in a cynaobacteria, Spirulina platensis and a microalga, Chlorella sp. 786. The study organisms were grown in both atmospheric (control sample, 0.035%) and high (exposed sample, 10%) $CO_2$ concentrations. Second dimension (2D) electrophoresis revealed a huge difference in the protein profiles of both organisms suggesting the induction of CCM related proteins in the sample maintained at atmospheric $CO_2$ concentration and the repression of CCM related proteins in the sample maintained at 10% $CO_2$. Liquid chromatography-mass spectroscopy analysis revealed the presence of two important $C_i$ transporter proteins in the control sample of S. platensis, namely ferredoxin-$NADP^+$ reductase and ATP binding cassette (ABC) transport system protein. These proteins were only expressed in the control sample and were downregulated or not expressed at all in the exposed sample. Consequently, this study conclusively proves that CCMs are only inducted at low $CO_2$ concentrations and are not functional at high $CO_2$ concentration.

• ALGAE
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• 제33권1호
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• pp.127-141
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• 2018

Low efficiency in microalgal biomass production was largely attributed to the low density of algal cell cultures. Though mutations that reduced the level of chlorophyll or pigment content increased efficiency of photon usage and thus the cell-culture density under high-illumination growth conditions (e.g., >$500{\mu}mol\;photon\;m^{-2}\;s^{-1}$), it was unclear whether algae could increase cell-culture density under low-illumination conditions (e.g., ${\sim}50{\mu}mol\;photon\;m^{-2}\;s^{-1}$). To address this question, we performed forward genetic screening in Chlamydomonas reinhardtii. A pool of >1,000 insertional mutants was constructed and subjected to continuous subcultures in shaking flasks under low-illumination conditions. Complexity of restriction fragment length polymorphism (RFLP) pattern in cultures indicated the degree of heterogeneity of mutant populations. We showed that the levels of RFLP complexity decreased when cycles of subculture increased, suggesting that cultures were gradually populated by high cell-culture density (HCD) strains. Analysis of the 3 isolated HCD mutants after 30 cycles of subcultures confirmed that their maximal biomass production was 50-100% higher than that of wild type under low-illumination. Furthermore, levels of chlorophyll content in HCD mutant strains were similar to that of wild type. Inverse polymerase chain reaction analysis identified the locus of insertion in two of three HCD strains. Molecular and transcriptomic analyses suggested that two HCD mutants were a result of the gain-of-function phenotype, both linking to the abnormality of mitochondrial functions. Taken together, our results demonstrate that HCD strains can be obtained through continuous subcultures under low illumination conditions.

• Journal of Microbiology and Biotechnology
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• 제29권12호
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• pp.1957-1968
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• 2019

Tetraselmis is a green algal genus, some of whose species are important in aquaculture as well as biotechnology. In algal culture, fluorescent lamps, traditional light source for culturing algae, are now being replaced by a cost-effective light-emitting diodes (LEDs). In this study, we investigated the effect of LED light of different wavelengths (white, red, yellow, and blue) on the growth of Tetraselmis suecica and its associated microbial community structures using the next-generation sequencing (NGS). The fastest growth rate of T. suecica was shown in the red light, whereas the slowest was in yellow. The highest OTUs (3426) were identified on day 0, whereas the lowest ones (308) were found on day 15 under red light. The top 100 OTUs associated with day 0 and day 5 cultures of T. suecica under the red and yellow LED were compared. Only 26 OTUs were commonly identified among four samples. The highest numbers of unique OTUs were identified at day 0, indicating the high degree of initial microbial diversity of the T. suecica inoculum. The red light-unique OTUs occupied 34.98%, whereas the yellow-specific OTUs accounted for only 2.2%. This result suggested a higher degree of interaction in T. suecica culture under the red light, where stronger photosynthesis occurs. Apparently, the microbial community associated with T. suecica related to the oxygen produced by algal photosynthesis. This result may expand our knowledge about the algae-bacteria consortia, which would be useful for various biotechnological applications including wastewater treatment, bioremediation, and sustainable aquaculture.

• 유기물자원화
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• 제8권2호
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• pp.71-76
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• 2000

미세조류를 이용한 생물학적 이산화탄소 고정화 방법은 이산화탄소를 효과적으로 제거할 수 있는 방법으로 널리 알려져 있다. 따라서 발전소 등에서 배출되는 폐탄산가스를 고정화하기 위하여 미세조류의 성장에 영향을 주는 운전 조건을 평가하였다. 본 실험에서 사용된 Euglena gracilis는 산성 조건에서도 잘 성장하여 오염의 가능성이 적고, 고농도 이산화탄소(10-20%)에 대한 내성이 강한 것으로 알려져 있다 또한, 단백질 함량이 풍부하고 세포벽이 얇아 배양 후 사료로의 이용성도 크다. 광반응기에서 회분식 배양을 이용하여 운전 조건을 평가한 결과 실험 조건에서 최적 pH는 3.5, 온도는 $27^{\circ}C$, 이산화탄소 농도는 5~10%, 광도는 $520{\mu}mol/m^2/s$로 각각 조사되었다. 또한 이러한 자료를 기초로 하여 연속 운전을 실시한 결과 $CO_2$농도 10%, HRT 4일에서 최종 건조 무게 $1.2g/{\ell}$로 높은 성장율을 얻었다.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1060-1066
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• 2005

Under environmental stress, such as strong irradiance or nitrogen deficiency, unicellular green algae of the genus Haematococcus accumulate secondary carotenoids, i.e. astaxanthin, in the cytosol. The induction and regulation of astaxanthin biosynthesis in microalgae has recently received considerable attention owing to the increasing use of secondary carotenoids as a source of pigmentation for fish aquacultures, and as a potential drug in cancer prevention as a free-radical quencher. Accordingly, this study generated expressed sequence tags (ESTs) from a library constructed from astaxanthin-induced Haematococcus pluvialis. Partial sequences were obtained from the 5' ends of 1,858 individual cDNAs, and then grouped into 1,025 non-overlapping sequences, among which 708 sequences were singletons, while the remainder fell into 317 clusters. Approximately $63\%$ of the EST sequences showed similarity to previously described sequences in public databases. H. pluvialis was found to consist of a relatively high percentage of genes involved in genetic information processing ($15\%$) and metabolism ($11\%$), whereas a relatively low percentage of sequences was involved in the signal transduction ($3\%$), structure ($2\%$), and environmental information process ($3\%$). In addition, a relatively large fraction of H. pluvialis sequences was classified as genes involved in photosynthesis ($9\%$) and cellular process ($9\%$). Based on this EST analysis, the full-length cDNA sequence for superoxide dismutase (SOD) of H. pluvialis was cloned, and the expression of this gene was investigated. The abundance of SOD changed substantially in response to different culture conditions, indicating the possible regulation of this gene in H. pluvialis.