• Title/Summary/Keyword: functional characterization

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Characterization of Chlorella Vulgaris Mutants Generated by EMS (Ethyl Methane Sulphonate) (EMS (Ethyl Methane Sulphonate) 처리에 의한 Chlorella Vulgaris 변이주 생성 및 특성 분석)

  • Kim, Ok Ju;Lee, Jae-Hwa
    • Applied Chemistry for Engineering
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    • v.26 no.3
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    • pp.265-269
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    • 2015
  • Chlorella vulgaris (C. vulgaris) is a spherical unicellular green algae and the diameter ranges from 2 to $10{\mu}m$. C. vulgaris possess nutritional excellence because it contains various functional materials including high protein contents, chlorophyll, carotenoid, and chlorella growth factor (CGF). In order to study effects of mutagen, ethyl methane sulphonate (EMS) was used as a chemical mutagen and some mutants could be obtained. We named 2 type mutants as E14 and E24 obtained after treating with EMS. In the cell growth, growth patterns of mutants were similar to those of the wild type. Chlorophyll contents of E14 and E24 increased up to 99 and 52%, respectively compared to those of the wild type. The carotenoid content of E14 increased to 7%, but the value of E24 decreased 5% compared to that of the wild type. For the lipid contents E24 increased to 23%, while E14 decreased 12% when compared to those of the wild type. As a result, there is no difference between the mutants and wild type in the cell growth, but considering that mutants contains more physiological materials than those of the wild type, we can expect the mutants of C. vulgaris could be used as important high added-value materials.

Isolation and Characterization of Pyrimidine Auxotrophs from the Hyperthermophilic Archaeon Sulfolobus acidocaldarius DSM 639 (Sulfolobus acidocaldarius 균주로부터 피리미딘 영양요구주의 분리 및 특성 연구)

  • Choi, Kyoung-Hwa;Cha, Jae-Ho
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1370-1376
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    • 2011
  • To study the functional genomic analysis of a crenachaeon Sulfolobus acidocaldarius, we have constructed an auxotrophic mutant based on pyrEF, which encodes the pyrimidine biosynthetic enzymes orotate phosphoribosyltransferase and orotidine-5'-monophosphate decarboxylase. S. acidocaldarius was shown to be sensitive to 5-fluoroorotic acid (5-FOA), which can be selected for mutations in pyrEF genes within a pyrimidine biosynthesis cluster. Spontaneous 5-FOA-resistant mutants by ultraviolet, KH1U and KH2U, were found to contain two point mutations and a frame shift mutation in pyrE, respectively. Mutations at these sites from KH1U and KH2U decreased the activity of orotate phosphoribosyltransferase encoded by the pyrE gene and blocked the degradation of 5-FOA into toxic 5-FOMP and 5-FUMP that kill the cells. Therefore, KH1U and KH2U were uracil auxotrophs. Transformation of Sulfolobus-Escherichia coli shuttle vector pC bearing pyrEF genes from S. solfataricus P2 into S. acidocaldarius mutant KH2U restored 5-FOA sensitivity and overcame the uracil auxotrophy. This study establishes an efficient genetic strategy towards the systematic knockout of genes in S. acidocaldarius.

Characterization of Arthrospira platensis Cultured in Nano-bubble Hydrogen Water (나노기포 수소수에서 배양한 Arthrospira platensis 특성 확인)

  • Seo, Ji-Hye;Choi, Soo-Jeong;Lee, Sang-Hoon;Lee, Jae-Hwa
    • Applied Chemistry for Engineering
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    • v.26 no.4
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    • pp.421-426
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    • 2015
  • Arthrospira platensis (A. platensis) has been used in various fields including dietary supplements as it contains a high protein content and large amounts of unsaturated fatty acids. In addition, it has some pigments such as phycocyanin, myxoxanthophyll and zeaxanthin and thus has been used as a food additive and antioxidant substance. Nano-bubble hydrogen is to dissolve more than the saturation solubility in water by injecting the hydrogen gas in the nano-bubble hydrogen water. The nano-bubbles are known to possess higher antioxidant properties in addition to anticancer effects. In this paper, Arthrospira platensis was cultured in both a normal medium with distilled water and nano-bubble hydrogen water medium and their properties were compared. The cell growth and the content of chlorophyll and carotenoid in the nano-bubble hydrogen water was 15% higher than that of the control. The level of phycocyanin in nano-bubble hydrogen water was also 7% higher than that of the control. However, there were little differences in the lipid content between the nano-bubble and control. To determine the content of the antioxidants, the level of flavonoid and polyphenol were measured. The level of flavonoid in nano-bubble hydrogen water was found to be more than 70% increased when comparing to that of the control, while the level of polyphenol was similar to each other.

Fabrication and characterization of a Flexible Polyethylene terephthalate (PET) Electrode based on Single-walled carbon nanotubes (SWNTs) (단일벽 탄소나노튜브를 이용한 플렉시블 폴리에틸렌테레프탈레이트 (PET) 전극의 제조와 특성)

  • Du, Jin Feng;Kim, Jang Hun;Kim, Yong Ryeol;Jeong, Hyeon Taek
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.3
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    • pp.587-592
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    • 2016
  • In this study, flexible acid treated single walled carbon nanotubes (A-SWCNTs) electrodes were fabricated by using gold coated PET substrate and spray coating technique. The acid-treatment method was conducted to introduce functional groups on the SWCNTs wall, which could improve dispersability of the SWCNTs and its electrochemical property. The electrochemical properties of flexible A-SWCNTs electrode were carried out by cyclic voltammetry(CV), electrochemical impedance were carried out by cyclic voltammetry(CV), electrochemical impedance spectroscopy(EIS) and galvanostatic charge/discharge (GCD) cycles. As a results, The specific capacitance value of the unbent A-SWCNTs electrode was $67F{\cdot}g^{-1}$, which decreased to $63F{\cdot}g^{-1}$ (94% retention) after 1000 GCD cycles. Interestingly, the specific capacitance of the unbent A-SWCNTs electrode with application of the 1000 GCD cycles was retained even after 500 bending to $30^{\circ}$ with 6000 GCD cycles.

Characterization of Chlorella vulgaris Mutants Producing High Chlorophyll (클로로필 고생산성 Chlorella vulgaris 변이주의 특성 분석)

  • Park, Hyun-Jin;Kim, Ok Ju;Ha, Ji Min;Choi, Tae O;Lee, Jae-Hwa
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.275-279
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    • 2015
  • Micro-algae are unicellular photosynthetic organisms and produce pigments such as chlorophyll and carotenoid. Chlorella contains a lot of protein and functional components like lipids, chlorophyll and carotenoids. In this study we induced mutants of Chlorella vulgaris (C. vulgaris) through ultraviolet radiation (UV-B) and selected two mutants by pigment (chlorophyll and carotenoids) content. We named the mutants ‘UBM1-2’, ‘UBM2-57’ and they were cultivated for 21-days. Cell growth, dry cell weight, protein content, lipid and pigments content were measured. The results indicated that the mutants displayed slower cell growth, lower dry cell weight and protein content than the wild type. However, for UBM1-2 the lipid content was 21% higher than the wild type. In addition, the mutants’ chlorophyll content was 37% and 89% higher than the wild type and the carotenoids content was 27% and 70% higher than the wild type, respectively.

Isolation of a New Agar Degrading Bacterium, Maribacter sp. SH-1 and Characterization of its Agarase (신규 한천분해세균 Maribacter sp. SH-1의 분리 및 효소 특성조사)

  • Lee, Chang-Eun;Lee, Sol-Ji;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.156-162
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    • 2016
  • In this study, we isolated a new agar-degrading marine bacterium and characterized its agarase. An agardegrading marine bacterium SH-1 was isolated from seawater, collected from the seashore of Namhae in Gyeongnam province, Korea, and cultured in marine agar 2216 media. It was identified as Maribacter. sp. SH-1 by phylogenetic analyses, based on 16S rRNA gene sequence. The extracellular agarase was extracted from culture media of Maribacter sp. SH-1 and characterized. Its relative activities were 56, 62, 94, 100, and 8% at 20, 30, 40, 50, and 60℃, respectively, whereas 15, 100, 60, and 21% relative activities were observed at pH 5, 6, 7, and 8, respectively. Its extracellular agarase exhibited maximum activity (231 units/l) at pH 6.0 and 50℃, in 20 mM Tris-HCl buffer. Therefore, this agarase would be applicable as it showed the maximum activity at the temperature at which the agar is in a sol state. Furthermore, the agarase activities remained over 90% at 20, 30, and 40℃ after 0.5 h exposure at these temperatures. Thin layer chromatography analysis suggested that Maribacter sp. SH-1 produces extracellular β-agarase, as it hydrolyzes agarose to produce neoagarooligosaccharides, such as neoagarohexaose (34.8%), neoagarotetraose (52.2%), and neoagarobiose (13.0%). Maribacter sp. SH-1 and its β-agarase would be useful for the production of neoagarooligosaccharides, which shows functional properties, like skin moisturizing, skin whitening, inhibition of bacterial growth, and delay in starch degradation.

Physicochemical Characterization of Fermented Rhododendron micranthum Turcz. Extract and Its Biological Activity (꼬리진달래 발효추출물의 이화학적 특성 및 생리활성 연구)

  • Kim, Min-Jin;Yu, Sang-Mi;Kim, Do-Yeon;Heo, Tae-Im;Lee, Jun Woo;Park, Ji-Ae;Park, Chang-Su;Kim, Yeong-Su
    • Journal of Life Science
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    • v.28 no.8
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    • pp.938-944
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    • 2018
  • This study evaluated tyrosinase, elastase inhibitory, and antioxidant activities of fermented Rhododendron micranthum Turcz. extract using a lactic acid bacterium, Lactobacillus rhamnosus. The optimum conditions for fermentation of R. micranthum Turcz. extract were $37^{\circ}C$ and 3% R. micranthum Turcz. extract for 3 days based on the bacterial cell number, total phenolic compounds, DPPH radical scavenging activity, and tyrosinase and elastase inhibitory activity. After culture for 3 days using 3% R. micranthum Turcz. extract, the cell mass of L. rhamnosus reached $5.7{\times}10^9CFU/ml$. The results indicated that R. micranthum Turcz. extract can be used for industrial lactic acid bacteria culture. After fermentation under optimum conditions, the total content of phenolic compounds of the fermented R. micranthum Turcz. extract was 157 GAE mg/ml, and the $IC_{50s}$ of DPPH radical scavenging activity, ABTS radical scavenging activity, and tyrosinase and elastase inhibitory effects were 78.8, 79.8, 329.1, and $449.5{\mu}g/ml$, respectively. The fermented R. micranthum Turcz. extract exhibited 1.2-, 1.3-, 1.5-, 2.4-, and 5.6-fold higher total content of phenolic compounds, DPPH radical scavenging activity, ABTS radical scavenging activity, and tyrosinase and elastase inhibitory effects than the nonfermented R. micranthum Turcz. extract. These results indicated that fermented R. micranthum Turcz. extract using L. rhamnosus can be used for developing new natural functional ingredients for the health food or cosmetic industry.

Characterization of Cysteine Residues in Cabbage Phospholipase D by Sulfhydryl Group Modifying Chemicals (설프히드릴 변형 화합물질들에 의한 양배추 포스포리파제 D의 시스테인 잔기의 특성)

  • Go, Eun-Hui
    • Journal of the Korean Chemical Society
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    • v.50 no.5
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    • pp.362-368
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    • 2006
  • SH group modifying chemicals were used to characterize the eight cysteine residues of cabbage PLD. 5,5-dithiobis(2-nitrobenzoate)(DTNB) was used to titrate the SH group of cysteine residues . Based on the optical density at 412nm due to the reduced DTNB, 4 SH groups are found to be present in a native PLD while 8 SH groups in the denatured PLD whose tertiary structure was perturbed by 8M urea. The results imply that among the 8 cysteine residues of PLD, the half(4) are exposed on the surface whereas the other half are present at the interior of the enzyme tertiary structure. The PLD was inactivated by SH modifying reagents such as p-chloromercuribenzoate(PCMB), iodoacetate, iodoacetamide, and N-ethylmaleimide. At the addition of dithiothreitol(DTT) only the PCMB inhibited PLD activity was recovered reversibly. The micro-environment of the exposed SH group of cysteine residues was examined with various disulfide compounds with different functional groups and we found that anionic or neutral disulfides appear to be more effective than the positively charged cystamine for inactivating the PLD activity. The effect of redox state of cysteine residues on the PLD activity was further explored with H2O2. The oxidation of SH groups by H2O2 inhibited the PLD activity more than 70%, which was mostly recovered by DTT. From these results, we could confirm chemically that all the cysteine residues of PLD are present as in their reduced SH forms and the 4 SH groups exposed on the surface of the enzyme may play important roles in the regulation of PLD activity.

Characterization of Superoxide-dependent Endothelial Relaxing Factor(s) (Superoxide에 의존하여 내피세포에서 유리되는 이완성 물질의 특성에 대한 실험적 연구)

  • Lee, Gi-Nam;Lee, Won-Suk;Rhim, Byung-Yong;Hong, Ki-Whan
    • The Korean Journal of Pharmacology
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    • v.26 no.2
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    • pp.145-152
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    • 1990
  • We recently reported a development of an experimental system which can identify the release of a superoxide-dependent vasorelaxant factor from endothelial cells using a two-bath system. In the present work, we further exploited the above system and observed whether the superoxide-dependent relaxing factor(s), released from the porcine coronary artery (PCA) endothelium, was similar in relaxation to those obtained from cat thoracic aortic endothelium and cultured endothelial cells of bovine aorta. However, there was observed a novel difference among the former one and the latter two relaxing factors; the release of relaxing factor from PCA endothelium can be inhibited either by catalase or by superoxide dismutase (SOD), whereas the latter two can be inhibited only by SOD. It was further attempted to characterize the synthetic mechanisms of the relaxing factors: (1) They were readily inhibited by various lipoxygenase inhibitors (gossypol, nordihydroguaiaretic acid, AA 861, and eicosatetraynoic acid). (2) They were not inhibited by cyclooxygenase inhibitor (indomethacin) and by cytochrome P-450 monooxygenease inhibitors (proadifen and cimetidine). Thus, it is likely that these relaxing factors, although obtained from different species, show common functional roles of arteriolar relaxation. It is suggested that they are related to pathophysiological involvement of various tissue ischemia-reperfusion injuries.

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Effect of Genetic Predisposition on Blood Lipid Traits Using Cumulative Risk Assessment in the Korean Population

  • Go, Min-Jin;Hwang, Joo-Yeon;Kim, Dong-Joon;Lee, Hye-Ja;Jang, Han-Byul;Park, Kyung-Hee;Song, Ji-Hyun;Lee, Jong-Young
    • Genomics & Informatics
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    • v.10 no.2
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    • pp.99-105
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    • 2012
  • Dyslipidemia, mainly characterized by high triglyceride (TG) and low high-density lipoprotein cholesterol (HDL-C) levels, is an important etiological factor in the development of cardiovascular disease (CVD). Considering the relationship between childhood obesity and CVD risk, it would be worthwhile to evaluate whether previously identified lipid-related variants in adult subjects are associated with lipid variations in a childhood obesity study (n = 482). In an association analysis for 16 genome-wide association study (GWAS)-based candidate loci, we confirmed significant associations of a genetic predisposition to lipoprotein concentrations in a childhood obesity study. Having two loci (rs10503669 at LPL and rs16940212 at LIPC) that showed the strongest association with blood levels of TG and HDL-C, we calculated a genetic risk score (GRS), representing the sum of the risk alleles. It has been observed that increasing GRS is significantly associated with decreased HDL-C (effect size, $-1.13{\pm}0.07$) compared to single nucleotide polymorphism combinations without two risk variants. In addition, a positive correlation was observed between allelic dosage score and risk allele (rs10503669 at LPL) on high TG levels (effect size, $10.89{\pm}0.84$). These two loci yielded consistent associations in our previous meta-analysis. Taken together, our findings demonstrate that the genetic architecture of circulating lipid levels (TG and HDL-C) overlap to a large extent in childhood as well as in adulthood. Post-GWAS functional characterization of these variants is further required to elucidate their pathophysiological roles and biological mechanisms.