• 제목/요약/키워드: Metabolic Engineering

검색결과 744건 처리시간 0.094초

Dual Component Analysis for In Vivo T2* Decay of Hyperpolarized 13C Metabolites

  • Joe, Eunhae;Lee, Joonsung;Lee, Hansol;Yang, Seungwook;Choi, Young-Suk;Wang, Eunkyung;Song, Ho-Taek;Kim, Dong-Hyun
    • Investigative Magnetic Resonance Imaging
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    • 제21권1호
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    • pp.1-8
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    • 2017
  • Purpose: To investigate the exchange and redistribution of hyperpolarized $^{13}C$ metabolites between different pools by temporally analyzing the relative fraction of dual $T_2{^*}$ components of hyperpolarized $^{13}C$ metabolites. Materials and Methods: A dual exponential decay analysis of $T_2{^*}$ is performed for [1-$^{13}C$] pyruvate and [1-$^{13}C$] lactate using nonspatially resolved dynamic $^{13}C$ MR spectroscopy from mice brains with tumors (n = 3) and without (n = 4) tumors. The values of shorter and longer $T_2{^*}$ components are explored when fitted from averaged spectrum and temporal variations of their fractions. Results: The $T_2{^*}$ values were not significantly different between the tumor and control groups, but the fraction of longer $T_2{^*}$ [1-$^{13}C$] lactate components was more than 10% in the tumor group over that of the controls (P < 0.1). The fraction of shorter $T_2{^*}$ components of [1-$^{13}C$] pyruvate showed an increasing tendency while that of the [1-$^{13}C$] lactate was decreasing over time. The slopes of the changing fraction were steeper for the tumor group than the controls, especially for lactate (P < 0.01). In both pyruvate and lactate, the fraction of the shorter $T_2{^*}$ component was always greater than the longer $T_2{^*}$ component over time. Conclusion: The exchange and redistribution of pyruvate and lactate between different pools was investigated by dual component analysis of the free induction decay signal from hyperpolarized $^{13}C$ experiments. Tumor and control groups showed differences in their fractions rather than the values of longer and shorter $T_2{^*}$ components. Fraction changing dynamics may provide an aspect for extravasation and membrane transport of pyruvate and lactate, and will be useful to determine the appropriate time window for acquisition of hyperpolarized $^{13}C$ images.

알콜중독환자의 신경기능 장애: 생체 양성자 자기공명분광 연구 (Neuronal Dysfunction in Patients with Chronic Alcoholism Evaluated by In Vivo $^1H$ Magnetic Resonance Spectroscopy)

  • Bo-Young Choe;Euy-Neyng Kim;Chang-Wook Lee;In-Ho Baik;Kwang-Soo Lee;Byung-Chul Son;Heung-Jae Chun;Hyoung-Koo Lee;Tae-Suk Suh
    • Investigative Magnetic Resonance Imaging
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    • 제4권2호
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    • pp.94-99
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    • 2000
  • 목적 : 국한적 수분억제 생체내 양성자 자기공명분광학을 사용하여 만성 알코을 중독자와 정상 건강인의 뇌대사물질의 비율을 비교평가하였다. 대상 및 방법 : 만성 알코을 중독자 10명,정상 건강인 10명의 좌측 소뇌와 basal ganglia의 관심부위 $2{\times}2{\times}2{\;}\textrm{cm}^3$를 각각 선택하여 STEAM펄스시퀀스를 이용하였다 Creatine (Cr)을 기준으로 Marquart알고리즘을 이용하여 각 대사물질의 비율을 산출하였다. 결과 만성 알코을 중독자의 특징적인 소견은 정상인과 비교하여 볼 때 N-acetyl aspartate (NAA)/Cr비율이 감소하였다. 그러나 Choline (Cho)/Cr등 다른 대사비율은 정상인과 유이한 차이를 발견할수 없었다. NAA/Cr비율은 임상평가와 상당한 상호관계가 있는 것으로 나타났다. 결론 : 본 예비실험 결과는 만성 알코을 중독자의 NAA/Cr의 비율 감소를 토대로 환자 소뇌의 신경세포 소실을 추정할 수 있었다. NAA/Cr비율은 향후 만성 알코을 중독자의 임상평가를 추정할수 있는 대사표준으로 사용될 수 있음을 시사해주고 있다. 따라서 생체내 양성자 자기공명분광법은 양성자 비율을 토대로 만성 알코을 중독자의 임상평가에 유용한 방법으로 사용될 수 있으리라 사료된다.

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Translocalization of enhanced PKM2 protein into the nucleus induced by cancer upregulated gene 2 confers cancer stem cell-like phenotypes

  • Yawut, Natpaphan;Kaowinn, Sirichat;Cho, Il-Rae;Budluang, Phatcharaporn;Kim, Seonghye;Kim, Suhkmann;Youn, So Eun;Koh, Sang Seok;Chung, Young-Hwa
    • BMB Reports
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    • 제55권2호
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    • pp.98-103
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    • 2022
  • Increased mRNA levels of cancer upregulated gene (CUG)2 have been detected in many different tumor tissues using Affymetrix microarray. Oncogenic capability of the CUG2 gene has been further reported. However, the mechanism by which CUG2 overexpression promotes cancer stem cell (CSC)-like phenotypes remains unknown. With recent studies showing that pyruvate kinase muscle 2 (PKM2) is overexpressed in clinical tissues from gastric, lung, and cervical cancer patients, we hypothesized that PKM2 might play an important role in CSC-like phenotypes caused by CUG2 overexpression. The present study revealed that PKM2 protein levels and translocation of PKM2 into the nucleus were enhanced in CUG2-overexpressing lung carcinoma A549 and immortalized bronchial BEAS-2B cells than in control cells. Expression levels of c-Myc, CyclinD1, and PKM2 were increased in CUG2-overexpressing cells than in control cells. Furthermore, EGFR and ERK inhibitors as well as suppression of Yap1 and NEK2 expression reduced PKM2 protein levels. Interestingly, knockdown of β-catenin expression failed to reduce PKM2 protein levels. Furthermore, reduction of PKM2 expression with its siRNA hindered CSC-like phenotypes such as faster wound healing, aggressive transwell migration, and increased size/number of sphere formation. The introduction of mutant S37A PKM2-green fluorescence protein (GFP) into cells without ability to move to the nucleus did not confer CSC-like phenotypes, whereas forced expression of wild-type PKM2 promoted such phenotypes. Overall, CUG2-induced increase in the expression of nuclear PKM2 contributes to CSC-like phenotypes by upregulating c-Myc and CyclinD1 as a co-activator.

Heat Shock Causes Oxidative Stress and Induces a Variety of Cell Rescue Proteins in Saccharomyces cerevisiae KNU5377

  • Kim, Il-Sup;Moon, Hye-Youn;Yun, Hae-Sun;Jin, Ing-Nyol
    • Journal of Microbiology
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    • 제44권5호
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    • pp.492-501
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    • 2006
  • In this study, we attempted to characterize the physiological response to oxidative stress by heat shock in Saccharomyces cerevisiae KNU5377 (KNU5377) that ferments at a temperature of $40^{\circ}C$. The KNU5377 strain evidenced a very similar growth rate at $40^{\circ}C$ as was recorded under normal conditions. Unlike the laboratory strains of S. cerevisiae, the cell viability of KNU5377 was affected slightly under 2 hours of heat stress conditions at $43^{\circ}C$. KNU5377 evidenced a time-dependent increase in hydroperoxide levels, carbonyl contents, and malondialdehyde (MDA), which increased in the expression of a variety of cell rescue proteins containing Hsp104p, Ssap, Hsp30p, Sod1p, catalase, glutathione reductase, G6PDH, thioredoxin, thioredoxin peroxidase (Tsa1p), Adhp, Aldp, trehalose and glycogen at high temperature. Pma1/2p, Hsp90p and $H^+$-ATPase expression levels were reduced as the result of exposure to heat shock. With regard to cellular fatty acid composition, levels of unsaturated fatty acids (USFAs) were increased significantly at high temperatures ($43^{\circ}C$), and this was particularly true of oleic acid (C18:1). The results of this study indicated that oxidative stress as the result of heat shock may induce a more profound stimulation of trehalose, antioxidant enzymes, and heat shock proteins, as well as an increase in the USFAs ratios. This might contribute to cellular protective functions for the maintenance of cellular homeostasis, and may also contribute to membrane fluidity.

Transiently Experessed Salt-Stress Protection of Rice by Transfer of a Bacterial Gene, mtlD

  • Lee, Eun-A;Kim, Jung-Dae;Cha, Yoo-Kyung;Woo, Dong-Ho;Han, In-Seob
    • Journal of Microbiology and Biotechnology
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    • 제10권3호
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    • pp.415-418
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    • 2000
  • Productivity of a rice plant is greatly influenced by salt stress. One of the ways to achieve tolerance to salinity is to transfer genes encoding protective enzymes from other organisms, such as microorganisms. The bacterial gene, mtlD, which encodes mannitol-1-phosphate dehydrogenase (Mtl-DH), was introduced to the cytosol of a rice plant by an imbibition technique to overproduce mannitol. The germination and survival rate of the imbibed rice seeds were markedly increased by transferring the mtlD gene when it was delivered in either a pBIN19 or pBmin binary vector. When a polymerase chain reaction was performed with the genomic DNAs of the imbibed rice leaves as a template and with mtlD-specific primers, several lines were shown to contain an exogenous mtlD DNA. However, a reverse transcription (RT)-PCR analysis revealed that not all of them showed an expression of this foreign gene. This paper demonstrates that the growth and germination of rice plants transiently transformed with the bacterial gene, mtlD, are enhanced and these enhancements may have resulted from the experssion of the mtlD gene. The imbibition method empolyed in this study fulfills the requirements for testing the function of such a putative gene in vivo prior to the production of a stable transgenic plant.

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초음파 검사를 이용한 비알코올성 지방간에 운동이 미치는 영향 평가 (Evaluation of the Effect of Exercise on Nonalcoholic Fatty Liver By Sonography)

  • 김경연;임현수
    • 대한방사선기술학회지:방사선기술과학
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    • 제35권1호
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    • pp.17-23
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    • 2012
  • 비알코올성 지방간 질환(NAFLD)은 알코올을 과도하게 섭취하지 않고도 간세포 내에 지방이 축적되어 있는 상태이며, 이는 비만과 밀접한 관련이 있다고 연구되어 지고 있다. 이 연구의 목표는 NAFLD의 위험인자를 파악하여 예방하거나 위험요인을 관리하는데 있다. 이 연구는 2011년 5월 1일부터 10월 31일까지 6개월 동안 진행되었고, 복부초음파 검사를 시행한 83명 중 연구 실험에 적합한 11명을 특별히 심사해서 선정하였다. 연구결과는 다음과 같이 나타났는데 첫째, 체중과 체질량지수에 있어 유의한 감소경향을 나타냈고, 둘째, 초음파 진단 상 지방간 심화정도의 변화에서 유의한 감소경향을 나타났으며, 셋째, 칼로리에 상관없이 꾸준한 운동을 하면 지방간 심화정도가 감소경향을 나타냈다. 따라서, 이 연구에서는 장기적인 운동 프로그램을 적용할 경우에는 지방간 치료에 긍정적인 효과를 나타낼 수 있을 것으로 추론해 본다.

Importance of culture history on 17α-ethinylestradiol cometabolism by nitrifying sludge

  • Jantanaprasartporn, Angkana;Maneerat, Suppasil;Rongsayamanont, Chaiwat
    • Environmental Engineering Research
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    • 제23권1호
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    • pp.28-35
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    • 2018
  • $17{\alpha}-ethinylestradiol$ (EE2), a synthetic estrogen which interfere the endocrine and reproductive function in living organisms, has been found extensively to be deposited into municipal wastewater treatment plants and the environment via human excretion. EE2 has long been known to be efficiently cometabolized by ammonia-oxidizing bacteria (AOB) during ammonia ($NH_3$) oxidation. Current study aims to investigate the effect of culture history on the biotransformation of EE2 by nitrifying sludge which was enriched under different ammonia loading rates in continuous flow reactors. Result showed that past growth condition largely affected not only the metabolic rate of $NH_3$ oxidation but also EE2 cometabolism. Sludge previously acclimated with higher $NH_3$ loads as well as sludge dominated with AOB belong to high growth cluster (Nitrosomonas europaea-Nitrosococcus mobilis) showed higher rate of EE2 biotransformation than those one being acclimated with lower $NH_3$ loads because of its ability to provide more reducing power from $NH_3$ oxidation. Moreover, the correlation between the degradation rates of $NH_3$ and EE2 was higher in sludge being acclimated with higher load of $NH_3$ in comparison with other sludge. Implication of the findings emphasized the role of volumetric $NH_3$ loading rate in determining EE2 removal in wastewater treatment system.

고도불포화지방산 생합성: 식물에서의 대사공학적 응용 (Biosynthesis of Polyunsaturated Fatty Acids: Metabolic Engineering in Plants)

  • 김순희;김소연;김종범;노경희;김영미;박종석
    • Journal of Applied Biological Chemistry
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    • 제52권3호
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    • pp.93-102
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    • 2009
  • Polyunsaturated fatty acids (PUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have significantly beneficial effects on health in relation to cardiovascular, immune, and inflammatory conditions and they are involved in determining the biophysical properties of membranes as well as being precursors for signaling molecules. PUFA biosynthesis is catalyzed by sequential desaturation and fatty acyl elongation reactions. This aerobic biosynthetic pathway was thought to be taxonomically conserved, but an alternative anaerobic pathway for the biosynthesis of PUFA is now known to contain analogous polyketide synthases (PKS). Certain fish oil can be a rich source of PUFA although processed marine oil is generally undesirable as food ingredients because of the associated objectionable flavors that are difficult and cost-prohibitive to remove. Oil-seed plants contain only the 18-carbon polyunsaturated fatty acid alpha-linolenic acid, which is not converted in the human body to EPA and DHA. It is now possible to engineer common oilseeds which can produce EPA and DHA and this has been the focus of a number of academic and industrial research groups. Recent advances and future prospects in the production of EPA and DHA in oilseed crops are discussed here.

Production of tropane alkaloids by metabolic engineering of Hyoscyamus niger H6H(hyoscyamine $6{\beta}-hydroxylase$) gene introduced Scopolia parviflora hairy root

  • Kang, Young-Min;Lee, Ok-Sun;Jung, Hee-Young;Kim, Won-Jung;Kang, Seung-Mi;Min, Ji-Yun;Bahk, Dong-Jin;Yun, Dae-Jin;Bahk, Jung-Dong;Choi, Myung-Suk
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.568-570
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    • 2003
  • The Hyoscyamus niger hyoscyamine $6{\beta}-hydroxylase$ (H6H, EC 1.14.11.11) gene was introduced into the genome of a Scopolia parviflora by the binary vector system using the disarmed Agrobacterium rhizogenes strain KCTC 2703. Expression of H6H enzyme which are involved in alkaloids pathway by western blot analysis using proteins extracted from leaf, stem flower, branch root and main root were examined The enzyme expression was found only in the roots, with no expression in leaf, stem and flower. The alkaloids contents were the most higher in root and then leaf and stem has very small amount of alkaloid contents were analyzed by HPLC. The expression level of H6H in transgenic plants were two or more times than wild type plants. In transgenic plant which constitutively expresses H6H enzyme, high concentration of scopolamine was accumulated.

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형질전환된 각질형성세포에서 생약추출물에 의한 $NF-{\kappa}B$ 활성화 억제효과 탐색 (Screening of Crude Drugs for the Inhibitory Effect on $NF-{\kappa}B$ Activation in Transfectant HaCaT Cells)

  • 안광석;김성기;문기영;한범수;강삼식;김영식
    • 생약학회지
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    • 제34권2호통권133호
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    • pp.156-160
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    • 2003
  • $NF-{\kappa}B$ (nuclear factor-kappa B) plays a particularly central role in epidermal biology. It has been established that ultraviolet radiation (UVR) is one of the mechanisms to induce the activation of $NF-{\kappa}B$ in human skin. We previously demonstrated that melanogenic inhibitors may act through the inhibition of $NF-{\kappa}B$ activation in keratinocytes. In order to find another type of melanogenic inhibitors of $NF-{\kappa}B$ activation, various kinds of the extracts from crude drugs $(30\;{\mu}g/ml)$ were preincubated with transfectant HaCaT cells for 3 hrs and then UVR $(60\;mj/cm^2)$ was irradiated. UVR-exposed cells were incubated for another 6 hrs to measure the $NF-{\kappa}B$ activity. $NF-{\kappa}B$ activation was measured with the secreatory alkaline phosphates (SEAP) reporter gene assay using a fluorescence detection method. Among natural products, Lycium chinense, Acanthopanax senticosus, Angelica koreana, Kalopanax pictus and Asparagus cochinchinensis were the most potent inhibitors of $NF-{\kappa}B$ activation by UVR. These observations suggest that some crude drugs might act partially through the modulation of the synthesis of melanotrophic factors to decrease melanogenesis in keratinocytes.