• Title/Summary/Keyword: Novel engineering

Search Result 8,184, Processing Time 0.039 seconds

Metabolic Engineering of the Brassinosteroid Biosynthetic Pathways (Brassinosteroid의 대사공학)

  • Lee, Mi-Ock;Song, Ki-Hong;Lee, Hyun-Kyung;Jung, Ji-Yoon;Choe, Vit-Nary;Choe, Sung-Hwa
    • Journal of Plant Biotechnology
    • /
    • v.29 no.2
    • /
    • pp.139-144
    • /
    • 2002
  • Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd /dwf3 were shown to be blocked in D$^4$reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bril/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus it is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRIl could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

Ginseng Gintonin Activates the Human Cardiac Delayed Rectifier K+ Channel: Involvement of Ca2+/Calmodulin Binding Sites

  • Choi, Sun-Hye;Lee, Byung-Hwan;Kim, Hyeon-Joong;Jung, Seok-Won;Kim, Hyun-Sook;Shin, Ho-Chul;Lee, Jun-Hee;Kim, Hyoung-Chun;Rhim, Hyewhon;Hwang, Sung-Hee;Ha, Tal Soo;Kim, Hyun-Ji;Cho, Hana;Nah, Seung-Yeol
    • Molecules and Cells
    • /
    • v.37 no.9
    • /
    • pp.656-663
    • /
    • 2014
  • Gintonin, a novel, ginseng-derived G protein-coupled lysophosphatidic acid (LPA) receptor ligand, elicits $[Ca^{2+}]_i$ transients in neuronal and non-neuronal cells via pertussis toxin-sensitive and pertussis toxin-insensitive G proteins. The slowly activating delayed rectifier $K^+$ ($I_{Ks}$) channel is a cardiac $K^+$ channel composed of KCNQ1 and KCNE1 subunits. The C terminus of the KCNQ1 channel protein has two calmodulin-binding sites that are involved in regulating $I_{Ks}$ channels. In this study, we investigated the molecular mechanisms of gintonin-mediated activation of human $I_{Ks}$ channel activity by expressing human $I_{Ks}$ channels in Xenopus oocytes. We found that gintonin enhances $I_{Ks}$ channel currents in concentration- and voltage-dependent manners. The $EC_{50}$ for the $I_{Ks}$ channel was $0.05{\pm}0.01{\mu}g/ml$. Gintonin-mediated activation 1 of the $I_{Ks}$ channels was blocked by an LPA1/3 receptor antagonist, an active phospholipase C inhibitor, an $IP_3$ receptor antagonist, and the calcium chelator BAPTA. Gintonin-mediated activation of both the $I_{Ks}$ channel was also blocked by the calmodulin (CaM) blocker calmidazolium. Mutations in the KCNQ1 $[Ca^{2+}]_i$/CaM-binding IQ motif sites (S373P, W392R, or R539W)blocked the action of gintonin on $I_{Ks}$ channel. However, gintonin had no effect on hERG $K^+$ channel activity. These results show that gintonin-mediated enhancement of $I_{Ks}$ channel currents is achieved through binding of the $[Ca^{2+}]_i$/CaM complex to the C terminus of KCNQ1 subunit.

A Missense Mutation in Exon 5 of the Bovine Growth Hormone Gene (소 성장호르몬 유전자의 Exon 5번에서의 새로운 다형성 연구)

  • Yoon, D. H.;Kim, T. H.;Lee, K. H.;Park, E. W.;Lee, H. K.;Cheong, I. C.;Hong, K. C.
    • Journal of Animal Science and Technology
    • /
    • v.45 no.1
    • /
    • pp.13-22
    • /
    • 2003
  • Growth Hormone (GH) gene is a member of gene family through the evolutionary process from a small common ancestral gene by a series of gene duplications. The role of the GH in growth and performance controls has been extensively studied in human, mice and livestock. Many researchers have considered GH as a strong candidate gene for evaluation of genetic polymorphisms that could be associated with economic traits in cattle. We report here a novel missense mutation within the exon 5 of the bovine Growth Hormone (bGH) gene. We could amplified 522 bp fragments from eight unrelated Hanwoo cattle by PCR, then, subsequently cloned and sequenced. An Msp I RFLP corresponding to a C to T transition was observed at position 2258 nt. From this result, we could predict a missense mutation (Arg to Trp) at codon 166 in a highly conserved region among many mammals. Codominant Mendelian segregation of the two alleles, Msp I (+) and Msp I (-), was observed in two full-sib F2 families (n = 32, African taurine Bos taurus ${\times}$ African zebu Bos indicus) and eight half-sib Hanwoo families. For the availability of genetic marker, we have performed PCR-RFLP with a large number of individual animals from 15 different cattle breeds (European and Asian taurines, and African indicines). Consideration of breed frequencies of Msp I (-) allele in relation to breed type and their geographic origins, shows higher frequencies in humped breeds or Asian cattle breeds than in humpless or European breeds. This result indicates that the missense mutation can be contributed the functional significance such as the signal transduction through the receptor binding, also may be used as a marker for selection of the economic traits in Hanwoo.

Water Scarcity Assessment Using Green and Blue Water Concepts (그린워터 및 블루워터를 이용한 물부족 평가)

  • Kim, Sung Eun;Lee, Dong Kun;Yang, Byung Sun;Jin, Yihua
    • Journal of Environmental Impact Assessment
    • /
    • v.27 no.3
    • /
    • pp.267-278
    • /
    • 2018
  • With climate change and population growth, there are significant increases in water scarcity. There have been water security assessments to abate the gap between water demand and availability to support water resource management. However, most of the assessments are focusing on the water that flows through either on or below the land surface, failing to consider water that infiltrates and can be used by vegetation. This study presents water scarcity assessment accounting for Blue and Green water concept, and applied the method to Boryung region. Monthly streamflow, evapotranspiration, and soil moisture were estimated by SWAT modeling, and each of them was used to analyze Blue and Green water scarcity. Blue and Green water scarcity had different aspect, and the result indicated the time when water scarcity is more likely to happen. The water scarcity assessment framework presented in this paper provides novel assessment method integrating hydrologic and ecosystem aspects, thereby improving the understanding of how water resources should be managed.

Pharmacological Characterization of KR-31125, a Novel Nonpeptide AT1 Receptor Antagonist (안지오텐신 수용체 길항제 KR-31125의 특성에 관한 연구)

  • Lee, Sung-Hou
    • Journal of Life Science
    • /
    • v.20 no.6
    • /
    • pp.831-837
    • /
    • 2010
  • KR-31125 (2-butyl-5-dimethoxymethyl-6-phenyl-7-methyl-3-[[2'-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-3H-imidazo[4,5-b]pyridine) is a potent inhibitor of angiotensin II type 1 ($AT_1$) receptors in human recombinant $AT_1$ receptors and rabbit aorta. These in vitro studies revealed that KR-31125 inhibited specific [$^{125}I$] [$Sar^1$, $Ile^8$]-angiotensin II binding to human recombinant $AT_1$ receptors in a concentration dependent manner with an $IC_{50}$ value of $19.72{\pm}2.65$ nM. However, no interaction with $AT_2$ receptors was detected as displayed by the competition binding of [$^{125}I$] CGP 42112A to human recombinant $AT_2$ receptor. The binding action was also confirmed as a competitive mode that was identical to the previously studied compound, losartan. In addition, KR-31125 caused a nonparallel shift to the right in the concentration response curves to angiotensin II with a 30-80% decrease in the maximum contractile responses ($pK_B$: 7.63). Compared to the previous studies with losartan that showed a parallel right shift in the maximum contractile responses to AII ($pA_2$: 7.59), KR-31125 presented a different mode of action with a similar potency to losartan. These results demonstrate that KR-31125 is a highly potent and $AT_1$ selective angiotensin II receptor antagonist that can be applied to the fields of new diagnostic and research tools with upcoming in vivo study results.

Effects of gintonin on the proliferation, migration, and tube formation of human umbilical-vein endothelial cells: involvement of lysophosphatidic-acid receptors and vascular-endothelial-growth-factor signaling

  • Hwang, Sung-Hee;Lee, Byung-Hwan;Choi, Sun-Hye;Kim, Hyeon-Joong;Won, Kyung Jong;Lee, Hwan Myung;Rhim, Hyewon;Kim, Hyoung-Chun;Nah, Seung-Yeol
    • Journal of Ginseng Research
    • /
    • v.40 no.4
    • /
    • pp.325-333
    • /
    • 2016
  • Background: Ginseng extracts are known to have angiogenic effects. However, to date, only limited information is available on the molecular mechanism underlying the angiogenic effects and the main components of ginseng that exert these effects. Human umbilical-vein endothelial cells (HUVECs) are used as an in vitro model for screening therapeutic agents that promote angiogenesis and wound healing. We recently isolated gintonin, a novel ginseng-derived lysophosphatidic acid (LPA) receptor ligand, from ginseng. LPA plays a key role in angiogenesis and wound healing. Methods: In the present study, we investigated the in vitro effects of gintonin on proliferation, migration, and tube formation of HUVECs, which express endogenous LPA1/3 receptors. Results: Gintonin stimulated proliferation and migration of HUVECs. The LPA1/3 receptor antagonist, Ki16425, short interfering RNA against LPA1 or LPA3 receptor, and the Rho kinase inhibitor, Y-27632, significantly decreased the gintonin-induced proliferation, migration, and tube formation of HUVECs, which indicates the involvement of LPA receptors and Rho kinase activation. Further, gintonin increased the release of vascular endothelial growth factors from HUVECs. The cyclooxygenase-2 inhibitor NS-398, nuclear factor kappa B inhibitor BAY11-7085, and c-Jun N-terminal kinase inhibitor SP600125 blocked the gintonin-induced migration, which shows the involvement of cyclooxygenase-2, nuclear factor kappa B, and c-Jun N-terminal kinase signaling. Conclusion: The gintonin-mediated proliferation, migration, and vascular-endothelial-growth-factor release in HUVECs via LPA-receptor activation may be one of in vitro mechanisms underlying ginsenginduced angiogenic and wound-healing effects.

Optical Design of the Integrated Triplexer Fabricated by Micro Block Stacking Method (MBS 방법으로 제작한 집적형 Triplexer의 광학 설계)

  • Yoon, Hyun-Jae;Kim, Jong-Hyuk
    • Korean Journal of Optics and Photonics
    • /
    • v.22 no.4
    • /
    • pp.191-197
    • /
    • 2011
  • In this paper, we have designed an integrated triplexer which is the basic component for a FTTH(Fiber To The Home) system which can transmit CATV and voice/data at the same time in a single fiber. The integrated triplexer can be fabricated with a novel technique of "Micro-Block Stacking (MBS)" method which automatically aligns the optical components in the optical beam pass using accurate ceramic holders. We analyze the displacement of the optical focus according to the tolerances of the component dimensions and the assembly process using code V simulator. For the transmitter, the most serious shift of the focal points is caused by the displacements of the LD spot. So the focal point moves up to $72{\mu}m$ from the center point for ${\pm}25{\mu}m$, ${\pm}25{\mu}m$, ${\pm}30{\mu}m$ displacements. For the receiver the most serious shift of the focal points is caused by the displacements of a 0.8mm ball lens (for the analog receiving part) and a micro ball lens (for the digital receiving part), and the focal point moves up to $55{\mu}$ for ${\pm}55{\mu}m$, ${\pm}5{\mu}m$, ${\pm}55{\mu}m$ micro ball lens displacements.

Purification and Properties of a Novel Extracellular Agarase from Marine Bacterium, Sphingomonas paucimobilis AS-1 (해양미생물 Sphingomonas paucimobilis AS-1이 생산하는 새로운 extracelluar agarase의 정제 및 특성)

  • Jung, Il-Sun;Kim, Yu-Jung;Song, Hyo-Ju;Gal, Sang-Wan;Choi, Young-Ju
    • Journal of Life Science
    • /
    • v.18 no.1
    • /
    • pp.103-108
    • /
    • 2008
  • An agar-degrading marine bacterium, strain AS-1 was isolated from the seawater. The strain AS-1 was identified as Sphingomonas paucimobilis (90% probability) by VITEK. The optimum medium for agarase activity of the isolated strain was determined to be marine medium, marine broth 2216 containing 0.1% agar as carbon source. An extracellular agarase was purified 104-fold from the culture supernatant by ammonium sulfate precipitation, ion exchange chromatography and gel filtration methods. The molecular weight of the purified enzyme was estimated to be 80 kDa by SDS-PAGE. The optimum pH and temperature for activity were 7.0 and $40^{\circ}C$, respectively. Antioxidative activity of the strain AS- was 72% in the supernatant cultured for 12 h. The culture supernatant of the strain AS-1 showed antibacterial activity against bacteria causing putrefaction and food poisoning such as Escherichia coli, Staphylococcus aureus and Proteus vulgaris. However, the cell growth of the lactic aicd forming strain, Lactobacillus plantarium was promoted by the treatment of 10% culture supernatant of an agar-degrading strain.

Novel Sulfonated Poly(arylene ether sulfone) Composite Membranes Containing Tetraethyl Orthosilicate (TEOS) for PEMFC Applications (고분자 전해질형 연료전지를 위한 TEOS가 함유된 술폰화 폴리아릴렌에테르술폰 복합막의 제조 및 특성)

  • Lee, Keun-Kyu;Kim, Tae-Ho;Hwang, Taek-Sung;Hong, Young-Taik
    • Membrane Journal
    • /
    • v.20 no.4
    • /
    • pp.278-289
    • /
    • 2010
  • A series of composite membranes based on sulfonated poly(arylene ether sulfone) (SPAES) were prepared via addition of tetraethyl orthosilicate (TEOS) and solution casting method. The morphological structure, water uptake, proton conductivity of the resulting composite membranes were extensively investigated as function of the content of TEOS. By the sol-gel reaction, TEOS molecules were not completely converted to $SiO_2$ particles, but formed only oligomer-type. Also, EDS confirms that the resulting silicon dioxide was homogeneously distributed in the composite membranes. As the content of TEOS increased, the prepared membranes increased water uptake and proton conductivity at high temperature and low relative humidity condition. In particular, considerably high proton conductivity (0.015 S/cm) at $120^{\circ}C$ and 48%RH was demonstrated in the composite membrane containing 200% TEOS, which is 10 times greater than that of unmodified SPAES membrane. Also, the composite membranes were found to have enhanced thermal stability compared to the unmodified membrane.

Siamese Crocodile White Blood Cell Extract Inhibits Cell Proliferation and Promotes Autophagy in Multiple Cancer Cell Lines

  • Phosri, Santi;Jangpromma, Nisachon;Chang, Leng Chee;Tan, Ghee T.;Wongwiwatthananukit, Supakit;Maijaroen, Surachai;Anwised, Preeyanan;Payoungkiattikun, Wisarut;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
    • /
    • v.28 no.6
    • /
    • pp.1007-1021
    • /
    • 2018
  • Cancer represents one of the most significant threats to human health on a global scale. Hence, the development of effective cancer prevention strategies, as well as the discovery of novel therapeutic agents against cancer, is urgently required. In light of this challenge, this research aimed to evaluate the effects of several potent bioactive peptides and proteins contained in crocodile white blood cell extract (cWBC) against LU-1, LNCaP, PC-3, MCF-7, and CaCo-2 cancer cell lines. The results demonstrate that 25, 50, 100, and $200{\mu}g/ml$ cWBC exhibits a strong cytotoxic effect against all investigated cell lines ($IC_{50}$ $70.34-101.0{\mu}g/ml$), while showing no signs of cytotoxicity towards noncancerous Vero and HaCaT cells. Specifically, cWBC treatment caused a significant reduction in the cancerous cells' colony forming ability. A remarkable suppression of cancerous cell migration was observed after treatment with cWBC, indicating potent antimetastatic properties. The mechanism involved in the cancer cell cytotoxicity of cWBC may be related to apoptosis induction, as evidenced by typical apoptotic morphology features. Moreover, certain cWBC concentrations induced significant overproduction of ROS and significantly inhibited the $S-G_2/M$ transition in the cancer cell. The molecular mechanisms of cWBC in apoptosis induction were to decrease Bcl-2 and XIAP expression levels and increase the expression levels of caspase-3, caspase-8, and p53. These led to a decrease in the expression level of the cell cycle-associated gene cyclin-B1 and the arrest of cell population growth. Consequently, these findings demonstrate the prospect of the use of cWBC for cancer therapy.