• ALGAE
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• 제32권4호
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• pp.359-377
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• 2017

Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata is one of the most commercially useful macroalgae cultivated in southeastern China. In red algae, the biosynthesis of terpenoids through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway can produce a direct influence on the synthesis of many biologically important metabolites. In this study, two genes of cDNAs, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and 1-deoxy-D-xylulose-5-phosphate reductase (DXR), which encoding the first two rate-limiting enzymes among MEP pathway were cloned from P. haitanensis. The cDNAs of P. haitanensis DXS (PhDXS) and DXR (PhDXR) both contained complete open reading frames encoding polypeptides of 764 and 426 amino acids residues, separately. The expression analysis showed that PhDXS was significant differently expressed between leafy thallus and conchocelis as PhDXR been non-significant. Additionally, expression of PhDXR and its downstream gene geranylgeranyl diphosphate synthase were both inhibited by fosmidomycin significantly. Meanwhile, we constructed types of phylogenetic trees through different algae and higher plants DXS and DXR encoding amino acid sequences, as a result we found tree clustering consequences basically in line with the "Cavalier-Smith endosymbiotic theory." Whereupon, we speculated that in red algae, there existed only complete MEP pathway to meet needs of terpenoids synthesis for themselves; Terpenoids synthesis of red algae derivatives through mevalonate pathway came from two or more times endosymbiosis of heterotrophic eukaryotic parasitifer. This study demonstrated that PhDXS and PhDXR could play significant roles in terpenoids biosynthesis at molecular levels. Meanwhile, as nuclear genes among MEP pathway, PhDXS and PhDXR could provide a new way of thinking to research the problem of chromalveolata biological evolution.

• 통합자연과학논문집
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• 제4권3호
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• pp.202-209
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• 2011

Tuberculosis is a major health problem in humans because of its multidrug resistance and discovering new treatments for this disease is urgently required. The synthesis of isoprenoids in Mycobacterium tuberculosis has been reported as an interesting pathway to target. In this context, 2C-methyl-D-erythritol 4-phosphate (MEP) pathway of M. tuberculosis has drawn attention. The MEP pathway begins with the condensation of glyceraldehyde 3-phosphate and pyruvate forming 1-deoxy-D-xylulose 5-phosphate (DXP) which is catalyzed by 1-deoxy-D-xylulose 5-phosphate synthase (DXS). As there is no X-ray structure was reported for this target, comparative modeling was used to generate the three dimensional structure. The structure was further validated by PROCHECK, VERIFY-3D, PROSA, ERRAT and WHATIF. Molecular docking studies was performed with the substrate (Thiamine pyrophosphate) and the reported inhibitor 2-methyl-3-(4-fluorophenyl)-5-(4-methoxy-phenyl)-4H-pyrazolol[1,5-a]pyrimidin-7-one) against the developed model to identify the crucial residues in the active site. This study may further be useful to provide structure based drug design.

• BMB Reports
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• 제40권6호
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• pp.911-920
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• 2007

Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the leading infectious diseases to humans. It is urgent to discover novel drug targets for the development of antitubercular agents. The 2-C-methyl-Derythritol-4-phosphate (MEP) pathway for isoprenoid biosynthesis has been considered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammals. MEP cytidyltransferase (IspD), the third-step enzyme of the pathway, catalyzes MEP and CTP to form 4-diphosphocytidyl-2-C-methylerythritol (CDP-ME) and PPi. In the work, ispD gene from M. tuberculosis H37Rv (MtIspD) was cloned and expressed. With N-terminal fusion of a histidine-tagged sequence, MtIspD could be purified to homogeneity by one-step nickel affinity chromatography. MtIspD exists as a homodimer with an apparent molecular mass of 52 kDa. Enzyme property analysis revealed that MtIspD has high specificity for pyrimidine bases and narrow divalent cation requirements, with maximal activity found in the presence of CTP and $Mg^{2+}$. The turnover number of MtIspD is $3.4 s^{-1}$. The Km for MEP and CTP are 43 and $92{\mu}M$, respectively. Furthermore, MtIspD shows thermal instable above $50^{\circ}C$. Circular dichroism spectra revealed that the alteration of tertiary conformation is closely related with sharp loss of enzyme activity at higher temperature. This study is expected to help better understand the features of IspD and provide useful information for the development of novel antibiotics to treat M. tuberculosis.

• 대한임상검사과학회지
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• 제47권1호
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• pp.24-27
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• 2015

To measure motor evoked potentials (MEP) during emergency surgery is often difficult in patients with subarachnoid hemorrhage (SAH) from a ruptured cerebral aneurysm, The cause of these difficulties may be considered as damage to the motor pathway by hemorrhage. To identify the cause of difficulties in measuring MEP, we defined the association between motor evoked potentials during surgery and the severity of the hemorrhage in patients with subarachnoid hemorrhage.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2012년도 정기총회 및 춘계학술발표회
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• pp.20-20
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• 2012

Isoprenoids represent the most diverse group of metabolites, which are functionally and structurally identified in plant organism to date. Ginsenosides, glycosylated triterpenes, are considered to be the major pharmaceutically active ingredient of ginseng. Its backbones, categorized as protopanaxadiol (PPD), protopanaxatriol (PPT), and oleanane saponin, are synthesized via the isoprenoid pathway by cyclization of 2,3-oxidosqualene mediated with dammarenediol synthase or beta-amyrin synthase. The rate-limiting 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), which is the first committed step enzyme catalyzes the cytoplasmic mevalonate (MVA) pathway for isoprenoid biosynthesis. DXP reductoisomerese (DXR), yields 2-C-methyl-D-erythritol 4-phosphate (MEP), is partly involved in isoprenoid biosynthesis via plastid. Squalene synthase and squalene epoxidase are involved right before the cyclization step. The triterpene backbone then undergoes various modifications, such as oxidation, substitution, and glycosylation. Here we will discuss general biosynthesis pathway for the production of ginsenoside and its modification based on their subcellular biological functions.

• Journal of Yeungnam Medical Science
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• 제11권2호
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• pp.248-261
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• 1994

허혈성 뇌졸중 환자에서 운동장애의 판단 및 예후 판정에 운동유발전위 (motor evoked potential : MEP)의 유용성을 알아보기 위해 정상대조군 24례와 허혈성 뇌졸중 환자 24례를 대상으로 대뇌피질 및 척추부위에 자기자극을 가하며 단모지외전근 및 무지외전근에서 MEP를 기록하였고 각각의 근육에서 대뇌피질을 자극하여 기록된 MEP의 잠복기에서 척추부위를 자극하여 기록된 MEP의 잠복기를 감한 차이로 CMCT를 산출하였다. 대조군은 24례 모두에게 MEP를 유발할 수 있었고 환자군에서는 24폐 중 무반응을 나타낸 경우가 각 근육에서 11례였다. 정상대조군과 MEP를 유발할 수 있었던 뇌졸중 환자에서 단모지외전근에서의 평균 CMCT는 각각 $8.6{\pm}1.4$msec, $9.4{\pm}2.3$msec로 통계적 유의성은 없었고, 무지외전근에서는 $15.1{\pm}2.1$msec, $18.8{\pm}3.9$msec로 유의한 차이를 보였다(p<0.01). 허혈성 뇌졸중 환자에서 추체로 또는 중심피질 병변일 때 혹은 급성기의 운동장애가 심할 때 대뇌피질 자기자극에 무반응을 보인 경우가 많았고, 추체로를 제외한 피질하 병변일 때 혹은 경한 운동장애를 보일 때 CMCT가 정상인 경우가 많았다. 운동장애를 보였던 환자중 급성기의 CMCT가 정상인 경우는 1~2개월 후 상당한 운동회복을 보였고 대뇌피질 자기자극에 반응이 없을 때 운동회복은 미약하였다. 이로 미루어 MEP는 안전하고 간편한 중추성 운동신경계 전달경로의 전기생리학적 검사로써 뇌졸중환자의 운동장애를 판단하는 데 도움을 주고 급성기 운동장애의 예후를 예측하는 데 하나의 지표가 될 수 있을 것이라 사료된다.

• Journal of Microbiology and Biotechnology
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• 제30권5호
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• pp.762-769
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• 2020

Vitamin K2 (menaquinone) is an essential vitamin existing in the daily diet, and menaquinone-7 (MK-7) is an important form of it. In a recent work, we engineered the synthesis modules of MK-7 in Bacillus subtilis, and the strain BS20 could produce 360 mg/l MK-7 in shake flasks, while the methylerythritol phosphate (MEP) pathway, which provides the precursor isopentenyl diphosphate for MK-7 synthesis, was not engineered. In this study, we overexpressed five genes of the MEP pathway in BS20 and finally obtained a strain (BS20DFHG) with MK-7 titer of 415 mg/l in shake flasks. Next, we optimized the fermentation process parameters (initial pH, temperature and aeration) in an 8-unit parallel bioreactor system consisting of 300-ml glass vessels. Based on this, we scaled up the MK-7 production by the strain BS20DFHG in a 50-l bioreactor, and the highest MK-7 titer reached 242 mg/l. Here, we show that the engineered strain BS20DFHG may be used for the industrial production of MK-7 in the future.

• Journal of Plant Biotechnology
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• 제46권2호
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• pp.88-96
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• 2019

Terpenes constitute a large class of secondary metabolites in plants. The Oryza sativa terpene synthase is a vital gene in plant defense response. In this study, the molecular and physiological functions of Oryza sativa terpene synthase 30 (OsTPS30, LOC_Os08g07080) were investigated after exposure of the seeds and plants to gamma-rays. The OsTPS30 expression was slightly induced at 200 Gray (Gy), but was significantly induced at 400 Gy. The total terpenoid was synthesized more in OsTPS30-overexpressing (OX-OsTPS30) Arabidopsisthaliana plants than in wild-type (WT) plants. The OX-OsTPS30 plants exhibited resistance to gamma-rays, as compared to WT. The OX-OsTPS30 plants had significantly increased height and weight after gamma irradiation. Additionally, the activity of antioxidant enzymes was increased more in OX OsTPS30 plants than in WT plants after gamma irradiation. Furthermore, the OsTPS30-GFP fusion protein was mostly localized in the chloroplast, suggesting that OsTPS30 is putative MEP pathway-related terpene synthase.

• 대한한방내과학회지
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• 제28권3호
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• pp.434-441
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• 2007

Objectives : Peripheral neurodegeneration occurs in diabetes mellitus (DM), both sensory and motor nerve. but we don't know exactly if DM affects central nerve pathway for all studies. Electrophysiologic study is one of the most important diagnostic tools for diabetic neuropathy. Electroneurography and electromyography are usually used. but evoked potentials (EP) is more sensitive to small nerve fiber damages and useful for central nerve evaluation in addition to peripheral nerves. Most diabetic neuropathy studies by EP have been performed with somatosensory evoked potentials (SSEP). In contrast, the objective of this study is to investigate if DM targets central motor neurons by assessing the relation between fasting blood sugar (FBS) and motor evoked potentials (MEP) latency. Methods : We inspected the medical records of 34 patients who had MEP tests during admitting days. The latency from cervical portion to abductor pollicis brevis was used as peripheral motor conduction time (PMCT). and the latency from vertex to cervical portion was used as central motor conduction time (CMCT). Then, they were correlated to FBS using correlation analysis. Results : There was a significant linear relation between FBS and PMCT (Pearson's correlation coefficient r=0.487, p<0.01), but a poor linear relation between FBS and CMCT (Pearson's correlation coefficient r=-0.l97. p>0.05). Conclusions : This study suggests that prolonged latencies of MEP in DM may be due to peripheral neuropathy rather than dysfunction of central motor pathway. therefore the clinical use of MEP to diabetic neuropathy has to be divided segmentally.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1207-1215
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• 2014

Candida albicans, the major human fungal pathogen, undergoes morphological transition from the budding yeast form to filamentous growth in response to nitrogen starvation. In this study, we identified a new function of GST2, whose expression was required for filamentous growth of C. albicans under nitrogen-limiting conditions. The Gst2p showed Gst activity and required response to oxidative stress. The ${\Delta}gst2$ mutant displayed predominantly yeast phase growth in low ammonium media. Such morphological defect of ${\Delta}gst2$ mutants was not rescued by overexpression of Mep2p, Cph1p, or Efg1p, but was rescued by either overexpression of a hyperactive $RAS1^{G13V}$ allele or through exogenous addition of cyclic AMP. In addition, the ${\Delta}gst2$ mutants had lower levels of RAS1 transcripts than wild-type cells under conditions of nitrogen starvation. These results were consistent with the Ras1-cAMP pathway as a possible downstream target of Gst2p. These findings suggest that Gst2p is a significant component of nitrogen starvation-induced filamentation in C. albicans.