• 제목/요약/키워드: Metabolic Engineering

검색결과 749건 처리시간 0.028초

엽록체 항산화기구 대사조절에 의한 환경스트레스 내성 식물 (Transgenic Plants with Enhanced Tolerance to Environmental Stress by Metabolic Engineering of Antioxidative Mechanism in Chloroplasts)

  • 권석윤;이영표;임순;이행순;곽상수
    • Journal of Plant Biotechnology
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    • 제32권3호
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    • pp.151-159
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    • 2005
  • Injury caused by reactive oxygen species (ROS), known as oxidative stress, is one of the major damaging factors in plants exposed to environmental stress. Chloroplasts are specially sensitive to damage by ROS because electrons that escape from the photosynthetic electron transfer system are able to react with relatively high concentration of $O_2$ in chloroplasts. To cope with oxidative stress, plants have evolved an efficient ROS-scavenging enzymes such as superoxide dismutase (SOD) and ascorbate peroxidase (APX), and low molecular weight antioxidants including ascorbate, glutathione and phenolic compounds. To maintain the productivity of plants under the stress condition, it is possible to fortify the antioxidative mechanisms in the chloroplasts by manipulating the antioxidation genes. A powerful gene expression system with an appropriate promoter is key requisite for excellent stress-tolerant plants. We developed a strong oxidative stress-inducible peroxidase (SWPA2) promoter from cultured cells of sweetpotato (Ipomoea batatas) as an industrial platform technology to develop transgenic plants with enhanced tolerance to environmental stress. Recently, in order to develop transgenic sweetpotato (tv. Yulmi) and potato (Solanum tuberosum L. cv. Atlantic and Superior) plants with enhanced tolerance to multiple stress, the genes of both CuZnSOD and APX were expressed in chloroplasts under the control of an SWPA2 promoter (referred to SSA plants). As expected, SSA sweetpotato and potato plants showed enhanced tolerance to methyl viologen-mediated oxidative stress. In addition, SSA plants showed enhanced tolerance to multiple stresses such as temperature stress, drought and sulphur dioxide. Our results strongly suggested that the rational manipulation of antioxidative mechanism in chloroplasts will be applicable to the development of all plant species with enhanced tolerance to multiple environmental stresses to contribute in solving the global food and environmental problems in the 21st century.

Brassinosteroid의 대사공학 (Metabolic Engineering of the Brassinosteroid Biosynthetic Pathways)

  • 이미옥;송기홍;이현경;정지윤;최빛나리;최성화
    • Journal of Plant Biotechnology
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    • 제29권2호
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    • pp.139-144
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    • 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.

비만 환자에서 한방 비만 변증에 따른 방풍통성산과 방기황기탕의 치료효과 및 부작용 비교 : 무작위 배정, 이중 맹검, 위약-대조군 임상시험 (Efficacy and Adverse Events of Bangpungtongseong-san(Bofutsusho-san) and Bangkihwangki-tang(Boiogiot-tang) by Oriental Obesity Pattern Identification on Obese Subjects : Randomized, Double Blind, Placebo-controlled Trial)

  • 박정현;이명종;김호준;홍선우;이동기;유재욱;최선미;문진석;임지연;이정복
    • 한방재활의학과학회지
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    • 제21권2호
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    • pp.265-278
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    • 2011
  • Objectives : This trial were conducted to evaluate the efficacy and adverse events of Bofutsusho-san(BTS) and Boiogiot-tang(BOT) by oriental obesity pattern identification on obese subjects, as compared to placebo. Methods : 166 subjects(body mass index ${\geq}25kg/m^2$) were recruited and randomized to receive BT(n=55), FH(n=55) or placebo(n=56) for 8 weeks. Anthropometric factors, serum lipid, glucose, blood pressure(BP), pulse rate, resting metabolic rate and oriental obesity pattern identification questionnaire were measured at baseline and 8 weeks. Adverse events and safety outcome variables were also checked during trials. Results : The frequency of top-scored oriental obesity pattern was ordered by indigestion(食積) > stagnation of the liver qi(肝鬱) > yang deficiency(陽虛) > spleen deficiency(脾虛) > phlegm(痰飮) > blood stasis(瘀血) in subjects. BTS group significantly decreased body weight, body mass index(BMI), waist circumference(WC), body fat mass, total cholesterol and HDL-cholesterol in stagnation of the liver qi(肝鬱) and WC in indigestion(食積). BOT group showed significant decrease of body weight, BMI, WC, and body fat mass in indigestion(食積) not in deficiency(虛症). Adverse events were reported most frequently in yang deficiency(陽虛) by BT group and stagnation of the liver qi(肝鬱) by BOT group. Conclusions : Bofu-tsusho-san was effective in treating obesity with stagnation of the liver qi(肝鬱). Obesity pattern identification could be a useful diagnostic tool predicting treatment effects and adverse events.

Metabolic engineering of Vit C: Biofortification of potato

  • Upadhyaya, Chandrama P.;Park, Se-Won
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2010년도 정기총회 및 추계학술발표회
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    • pp.14-14
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    • 2010
  • Vitamin C (ascorbic acid) is an essential component for collagen biosynthesis and also for the proper functioning of the cardiovascular system in humans. Unlike most of the animals, humans lack the ability to synthesize ascorbic acid on their own due to a mutation in the gene encoding the last enzyme of ascorbate biosynthesis. As a result, vitamin C must be obtained from dietary sources like plants. In this study, we have developed two different kinds of transgenic potato plants (Solanumtuberosum L. cv. Taedong Valley) overexpressing strawberry GalUR and mouse GLoase gene under the control of CaMV 35S promoter with increased ascorbic acid levels. Integration of the these genes in the plant genome was confirmed by PCR and Southern blotting. Ascorbic acid(AsA) levels in transgenic tubers were determined by high-performance liquid chromatography(HPLC). The over-expression of these genes resulted in 2-4 folds increase in AsA intransgenic potato and the levels of AsA were positively correlated with increased geneactivity. The transgenic lines with enhanced vitamin C content showed enhanced tolerance to abiotic stresses induced by methyl viologen(MV), NaCl or mannitol as compared to untransformed control plants. The leaf disc senescence assay showed better tolerance in transgenic lines by retaining higher chlorophyll as compared to the untransformed control plants. Present study demonstrated that the over-expression of these gene enhanced the level of AsA in potato tubers and these transgenics performed better under different abiotic stresses as compared to untransformed control. We have also investigated the mechanism of the abiotic stress tolerance upon enhancing the level of the ascorbate in transgenic potato. The transgenic potato plants overexpressing GalUR gene with enhanced accumulation of ascorbate were investigated to analyze the antioxidants activity of enzymes involved in the ascorbate-glutathione cycle and their tolerance mechanism against different abiotic stresses under invitro conditions. Transformed potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed significant increase in the activities of superoxide dismutase(SOD, EC 1.15.1.1), catalase, enzymes of ascorbate-glutathione cycle enzymes such as ascorbate peroxidase(APX, EC 1.11.1.11), dehydroascorbate reductase(DHAR, EC 1.8.5.1), and glutathione reductase(GR, EC 1.8.1.7) as well as the levels of ascorbate, GSH and proline when compared to the untransformed tubers. The increased enzyme activities correlated with their mRNA transcript accumulation in the stressed transgenic tubers. Pronounced differences in redox status were also observed in stressed transgenic potato tubers that showed more tolerance to abiotic stresses when compared to untransformed tubers. From the present study, it is evident that improved to lerance against abiotic stresses in transgenic tubers is due to the increased activity of enzymes involved in the antioxidant system together with enhanced ascorbate accumulated in transformed tubers when compared to untransformed tubers. At moment we also investigating the role of enhanced reduced glutathione level for the maintenance of the methylglyoxal level as it is evident that methylglyoxal is a potent cytotoxic compound produced under the abiotic stress and the maintenance of the methylglyoxal level is important to survive the plant under stress conditions.

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고려홍삼에 함유된 선택조절제 및 항식욕감퇴 인자에 관한 연구 (Studies on Selective Modulators and Antianorexigenic Agents in Korean Red Ginseng)

  • Takaku Takeshi;Kameda Kenji;Matsuura Yukinaga;Sekiya Keizo;Okai Hideo;Okuda Hiromichi
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1988년도 학술대회지
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    • pp.28-32
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    • 1988
  • 에피네프린은 지방분해를 촉진하고 인슈린은 지방생성을 촉진한다는 것은 잘 알려져 있다. 최근에 한국홍삼분말 중에는 에피네프린으로 유도된 지방분해를 억제하며 인슐린이 매개하는 당으로부터의 지방합성을 촉진하는 아데노신과 산성펩티드 성분이 존재함이 밝혀졌다. 산성펩티드의 화학구조는 Glu-Glu-Glu-Glu-Glu-Glu-Glucose인 것으로 밝혀졌다. 한국홍삼으로 부터 추출된 이러한 물질들은 흰쥐 지방세포에서 반대의 대사경로를 통하여 지방분해를 억제하는 반면 지방합성을 촉진하기도 하는 선택적 조절작용을 나타낸다. 최근에 우리는 여러가지 악성종양을 가진 환자의 복수액으로부터 toxohormone-L로 명명된 독성물질을 분리하였다. Toxohormone-L은 흰쥐 지방세포에서 지방분해를 촉진하고 흰쥐에서 식욕감퇴를 유발했다. Toxohormone-L의 이러한 작용은 한국홍삼에서 분리된 진세노사이드 $Rb_2$에 의해서 억제됨이 발견되었다. 한국홍삼에서 발견된 이러한 물질의 생리학적 유의성을 언급하고자 한다.

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Aspergillus sp. PS-104의 액침배양증 mycellial pellet 형성에 미치는 황토의 영향 (Effects of Loess on the Mycellial Pellet Formation of Phosphate-solubilizing Fungus, Aspergillus sp. PS-104 in the Submerged Culture)

  • 강선철;구본성;태언희
    • 한국환경농학회지
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    • 제21권1호
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    • pp.17-23
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    • 2002
  • 유리인산 생산균을 생물비료화 하기 위한 노력의 일환으로 인산가용화능이 우수한 Aspergillus sp. PS-104 균주를 액침배양하면서 pellet 형성에 미치는 황토의 영향을 조사하였다 Aspergillus sp. PS-104의 분생포자를 황토를 포함하는 PDB 배지에 접종하여 50 rpm의 낮은 속도로 교반하여 배양하면 무정형의 불규칙적인 pellet을 형성하는 반면에 150 rpm의 높은 속도로 배양하면 구형의 규칙적인 pellet을 형성하였다. 또한 0$\sim$1.0% (W/V) 범위의 황토 첨가시에는 황토의 농도가 높을수록 pellet의 크기가 작게 형성되었으며, 1.0% 황토 농도에서 $1.0\pm0.1$ mm의 가장 작은 pellet이 형성되었다. 이 결과는 황토를 첨가하지 않는 경우에 비하여 약 4배 작아진 것이다. 그러나 황토 농도가 1.5% 이상 되면 무정형의 큰 pellet이 형성되었다. 또한 황토의 주성분인 $SiO_2$, $Fe_2O_3$, $Al_2O_3$, $CaCO_3$, $CaSO_4$ and $MgCO_3$ 분말을 각각 0$\sim$1.0% 농도로 배지에 첨가하여 pellet 형성에 미치는 영향을 조사한 결과 $Al_2O_3$를 제외하고는 첨가물의 농도가 높을수록 작은 크기의 pellet이 형성되었으며, $MgCO_3$ 를 첨가했을 때 가장 작은 크기의 pellet을 형성하였다.

Lnk is an important modulator of insulin-like growth factor-1/Akt/peroxisome proliferator-activated receptor-gamma axis during adipogenesis of mesenchymal stem cells

  • Lee, Jun Hee;Lee, Sang Hun;Lee, Hyang Seon;Ji, Seung Taek;Jung, Seok Yun;Kim, Jae Ho;Bae, Sun Sik;Kwon, Sang-Mo
    • The Korean Journal of Physiology and Pharmacology
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    • 제20권5호
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    • pp.459-466
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    • 2016
  • Adipogenic differentiation of mesenchymal stem cells (MSCs) is critical for metabolic homeostasis and nutrient signaling during development. However, limited information is available on the pivotal modulators of adipogenic differentiation of MSCs. Adaptor protein Lnk (Src homology 2B3 [SH2B3]), which belongs to a family of SH2-containing proteins, modulates the bioactivities of different stem cells, including hematopoietic stem cells and endothelial progenitor cells. In this study, we investigated whether an interaction between insulin-like growth factor-1 receptor (IGF-1R) and Lnk regulated IGF-1-induced adipogenic differentiation of MSCs. We found that wild-type MSCs showed greater adipogenic differentiation potential than $Lnk^{-/-}$ MSCs. An ex vivo adipogenic differentiation assay showed that $Lnk^{-/-}$ MSCs had decreased adipogenic differentiation potential compared with wild-type MSCs. Interestingly, we found that Lnk formed a complex with IGF-1R and that IGF-1 induced the dissociation of this complex. In addition, we observed that IGF-1-induced increase in the phosphorylation of Akt and mammalian target of rapamycin was triggered by the dissociation of the IGF-1R-Lnk complex. Expression levels of a pivotal transcription factor peroxisome proliferator-activated receptor gamma ($PPAR-{\gamma}$) and its adipogenic target genes (LPL and FABP4) significantly decreased in $Lnk^{-/-}$ MSCs. These results suggested that Lnk adaptor protein regulated the adipogenesis of MSCs through the $IGF-1/Akt/PPAR-{\gamma}$ pathway.

Box model을 이용한 서해 곰소만 하전 갯벌의 질소 수지 (Nitrogen Budget Analysis Using a Box Model for Hajeon Tidal Flat in the West Coast of Korea)

  • 유재원;홍재상;양성렬;박경
    • 한국해양학회지:바다
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    • 제7권4호
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    • pp.257-266
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    • 2002
  • 1999년 4월 17-18일 양일 간 전라북도 부안, 변산반도 남단에 위치한 곰소만의 하전 갯벌에서 3.0$\times$4.5 ㎦의 연구 지역을 설정 , box model을 이용한 물질(Chl-a, DIN, DON, TN) 수지 계산과 자정 기능의 추정을 위한 현장 조사를 실시하였다. 조사 지역의 Chl-a 소실률(-0.05mg Chl/$m^2$/hr)은 일본 아이치현 미카와만 이시키 갯벌의 7% 수준으로 나타났다. 이렇게 낮은 소실률은 낮은 식물플랑크톤 현존량, 조사 시기의 낮은 온도에 따른 대사율 감소 그리고 조사 지역의 낮은 대형저서동물량 때문인 것으로 추정되었다. 질소의 측면에서 물질 수지의 계산 결과, 13.5 $\textrm{km}^2$ 면적의 하전 갯벌에서는 하루 동안 TN및 PON의 소실률이 각각 8.36$\times$$10^{2}$, 5.36$\times$$10^{3}$kgN/day로 그리고 식물플랑크톤 소실에 따른 PON 소실률이 1.62$\times$$10^{2}$kg N/day로 추정되어 갯벌이 연안해역에서 질소 제거에 중요한 역할을 수행하고 있음을 시사하였다. 곰소만 하전 갯벌에서의 PON 소실률을 기존의 1개 하수처리장 평균 처리비용과 비교 할 때 13.5$\textrm{km}^2$ 면적의 하전 갯벌의 수질 정화기능은 대규모 하수처리장 2개 이상에 상당하는 경제적 가치를 지니는 것으로 나타났다

천연물 성분을 이용한 환경 유해미생물의 biofilm 생성 저해능 비교에 관한 연구 (Comparison of In vitro Anti-Biofilm Activities of Natural Plant Extracts Against Environment Harmful Bacteria)

  • 강은진;박지헌;진슬;김영록;도형기;양웅석;이재용;황철원
    • 한국환경과학회지
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    • 제28권2호
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    • pp.225-233
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    • 2019
  • In this study, we investigated the in vitro anti-biofilm activities of plant extracts of chives (Allium tuberosum), garlic (Allium sativum), and radish (Raphanus sativus L.) against environment harmful bacteria (gram-positive Staphylococcus aureus and, gram-negative Salmonella typhimurium and Escherichia coli O157:H7). In the paper disc assay, garlic extracts exhibited the highest anti-biofilm activity. The Minimal Inhibitory Concentration (MIC) of all plant extracts was generally higher for gram-negative bacteria than it was for gram-positive bacteria. Gram-negative bacteria were more resistant to plant extracts. The tetrazolium dye (XTT) assay revealed that, each plant extract exhibited a different anti-biofilm activity at the MIC value depending on the pathogen involved. Among the plant extracts tested, garlic extracts (fresh juice and powder) effectively reduced the metabolic activity of the cells of food-poisoning bacteria in biofilms. These anti-biofilm activities were consistent with the results obtained through light microscopic observation. Though the garlic extract reduced biofilm formation for all pathogens tested, to elucidate whether this reduction was due to antimicrobial effects or anti-biofilm effects, we counted the colony forming units of pathogens in the presence of the garlic extract and a control antimicrobial drug. The garlic extract inhibited the E. coli O157:H7 biofilm effectively compared to the control antimicrobial drug ciprofloxacin; however, it did not inhibit S. aureus biofilm significantly compared to ciprofloxacin. In conclusion, garlic extracts could be used as natural food preservatives to prevent the growth of foodborne pathogens and elongater the shelf life of processed foods.

주스착즙 방식에 따른 당근 주스의 품질 특성 변화 (Effect of extraction method on quality characteristics of the carrot juice)

  • 박혜정;김지윤;이송민;김희숙;이상현;이문현;장정수
    • 한국식품과학회지
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    • 제51권4호
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    • pp.369-378
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    • 2019
  • 본 연구는 원심분리형, 외기어 방식 그리고 쌍기어 방식과 같은 착즙 방법에 따른 당근 주스의 품질 특성을 비교하기 위해 수행되었다. 신선한 주스의 품질은 착즙률, 영양성분 및 층 분리 안정성을 통해 평가되었다. 쌍기어 착즙 방식은 가장 높은 착즙률을 보였다. 게다가 당근 주스의 미네랄 함량은 쌍기어 착즙방식으로 제조된 당근 주스에서 가장 높게 나타났으며, 특히 유일하게 쌍기어 방식의 착즙이 100 g의 당근으로부터 1일 권장 섭취량 이상의 ${\beta}$-carotene이 얻어졌다. 더욱이 당근 주스의 입자 크기가 가장 작게 나타난 것은 쌍기어 방식의 착즙이었고 다음으로 외기어 방식과 원심분리형으로 나타났다. 따라서 쌍기어 방식의 착즙을 최적의 방법으로 선정하고, 이를 이용해 당근 주스의 항산화 활성 및 대사 활성 변화를 조사하였다. 그 결과 당근 주스는 ${\alpha}$-tocopherol 보다 더 높은 지질과산화 저해율을 보였으며, 위장과 소장 소화를 거치면서 angiotensin I-converting enzyme(ACE) 저해 활성이 증가되었다.