• Preventive Nutrition and Food Science
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• 제3권1호
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• pp.36-42
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• 1998

Ion-selective eleltrodes(ISEs) are simple electrodechemical devices for the direct measurement of ions in the samples. A novel potentiometric biosensor for the determination of L-Malate or D-isocitrate has been developed by using CO2-3 -ISE-FIA system was composed of a pump, an injector, a malic enzyme or isocitric dehydrogenase enzyme reactor, a CO2-3 -ISE, a pH/mV meter, and an integrater. The various factors, such as buffer capacity types of plstericizer and polymer, were optimized for the CO2-3 selectivity. In this novel CO2-3 --ISE-FIA system, the potential difference due to the amount of CO2-3 produced from each enzyme reaction was proportional to the amount of L-malate or D-isocitrate.

• BMB Reports
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• 제32권4호
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• pp.385-392
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• 1999

The $NAD^+$-specific activity of a dual coenzyme-specific isocitrate dehydrogenase (IDH; EC 1.1.1.41) from the primitive fungus Pythium ultimum was investigated to elucidate the regulatory factors that may influence the intracellular distribution of carbon and the availability of intermediates, e.g. citrate, for fatty acid synthesis. Inhibition of $NAD^+$-IDH activity by diphospho- and triphosphonucleotides (ATP, ADP, and GTP) reflected the sensitivity of this enzyme to cellular energy charge even though monophosphonucleotides (AMP and GMP) had little effect on activity. NADPH, but not NADH, substantially inhibited $NAD^+$-IDH activity, showing noncompetitive inhibition with isocitrate. Oxalacetate and ${\alpha}$-ketoglutarate showed competitive inhibition with isocitrate, while citrate and cis-aconitate showed mixed-noncompetitive inhibition with isocitrate. Inhibition by these substances ranged from 29 to 46% at 10 mM. The inhibitory effect of oxalacetate was increased synergistically by glyoxylate, which alone caused 31% uncompetitive inhibition at 10 mM, and a mixture of the two substances at 1 mM each showed 98% inhibition of $NAD^+$-IDH activity. The regulation of $NAD^+$-IDH in Pythium ultimum seems to be a complex process involving mitochondrial metabolites. The addition of glyoxylate (3 mM) and oxalacetate (3 mM) to the culture medium resulted in the production of 49% more lipid by P. ultimum.

• Bulletin of the Korean Chemical Society
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• 제12권5호
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• pp.504-509
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• 1991

S. fradiae showed very high activity of isocitrate dehydrogenase compared to other microorganisms. The activity of this enzyme was increased with the growth of the organism. But the increase might not imply its involvement in the growth. Rather its increased activity seemed to have a connection with the biosynthesis of neomycin. The enzyme showed high specificity toward $NADP^+$ and D-isocitrate with Km values of 5.75 and 6.74 uM, respectively, It was activated by $Mn^{2+}$. Its molecular weight was estimated from its gel retardation coefficient to be in the range of 61,000-63,000 daltons and its optimum pH was 8.0. The enzyme was thermally unstable.

• Mycobiology
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• 제47권2호
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• pp.207-216
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• 2019

Talaromyces marneffei is the only dimorphic species in its genus and causes a fatal systemic mycosis named talaromycosis. Our previous study indicated that knockdown of AcuD gene (encodes isocitrate lyase of glyoxylate bypass) of T. marneffei by RNA interference approach attenuated the virulence of T. marneffei, while the virulence of the AcuD knockout strains was not studied. In this study, T. marneffei-zebrafish infection model was successfully established through hindbrain microinjection with different amounts of T. marneffei yeast cells. After co-incubated at $28^{\circ}C$, the increasing T. marneffei inoculum doses result in greater larval mortality; and hyphae generation might be one virulence factor involved in T. marneffei-zebrafish infection. Moreover, the results demonstrated that the virulence of the ${\Delta}AcuD$ was significantly attenuated in this Zebrafish infection model.

• Archives of Pharmacal Research
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• 제27권1호
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• pp.48-52
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• 2004

DA-11004 is a synthetic, potent NADP-dependent isocitrate dehydrogenase (IDPc) inhibitor where $IC_{50}$ for IDPc is 1.49 $\mu$M. The purpose of this study was to evaluate the effects of DA-11004 on the high fat high sucrose (HF)-induced obesity in male C57BL/6J mice. After completing a 8-week period of experimentation, the mice were sacrificed 1hr after the last DA-11004 treatment and their blood, liver, and adipose tissues (epididymal and retroperitoneal fat)were collected. There was a significant difference in the pattern of increasing body weight between the HF control and the DA-11004 group. In the DA-11004 (100 mg/kg) treated group the increase in body weight significantly declined and a content of epididymal fat and retroperitoneal fat was also significantly decreased as opposed to the HF control. DA-11004 (100 mg/kg) inhibited the IDPc activity, and thus, NADPH levels in plasma and the levels of free fatty acid (FFA) or glucose in plasma were less than the levels of the HF control group. In conclusion, DA-11004 inhibited the fatty acid synthesis in adipose tissues via IDPc inhibition, and it decreased the plasma glucose levels and FFA in HF diet-induced obesity of C57BL/6J mice.

• BMB Reports
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• 제54권12호
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• pp.601-607
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• 2021

Odorant receptors (ORs) account for about 60% of all human G protein-coupled receptors (GPCRs). OR expression outside of the nose has functions distinct from odor perception, and may contribute to the pathogenesis of disorders including brain diseases and cancers. Glioma is the most common adult malignant brain tumor and requires novel therapeutic strategies to improve clinical outcomes. Here, we outlined the expression of brain ORs and investigated OR expression levels in glioma. Although most ORs were not ubiquitously expressed in gliomas, a subset of ORs displayed glioma subtype-specific expression. Moreover, through systematic survival analysis on OR genes, OR51E1 (mouse Olfr558) was identified as a potential biomarker of unfavorable overall survival, and OR2C1 (mouse Olfr15) was identified as a potential biomarker of favorable overall survival in isocitrate dehydrogenase (IDH) wild-type glioma. In addition to transcriptomic analysis, mutational profiles revealed that somatic mutations in OR genes were detected in > 60% of glioma samples. OR5D18 (mouse Olfr1155) was the most frequently mutated OR gene, and OR5AR1 (mouse Olfr1019) showed IDH wild-type-specific mutation. Based on this systematic analysis and review of the genomic and transcriptomic profiles of ORs in glioma, we suggest that ORs are potential biomarkers and therapeutic targets for glioma.

• Clinical and Experimental Reproductive Medicine
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• 제34권1호
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• pp.19-31
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• 2007

목 적: 자궁내막증은 자궁내부에 존재하여야 할 자금내막조직이 자궁 외에 존재하는 질환으로 그 발생기전은 아직 명확하게 밝혀져 있지 않다. 이에 저자들은 자궁내막증 환자와 정상 대조군의 자궁내막조직 간의 유전자 발현의 차이가 자궁내막증의 발병과 관련이 있을 것이라는 가정 하에 DNA microarray 기술을 도입하여 연구를 시행하였다. 연구방법: 2002년 1월부터 2002년 12월까지의 기간 동안 본원 산부인과에서 자궁내막증 환자와 자궁내막증 이외의 다른 부인과적 질환으로 수술을 시행한 환자들을 대상으로 채취한 자궁내막 조직으로 KNU 4.8K cDNA chip을 이용하여 유전자 발현을 비교 연구하였다. 유전자칩으로 자궁내막증 조직에서 발현의 증감을 보였던 유전자 중에서 8종의 유전자를 대상으르 RT-PCR이나 real time RT-PCR 법을 통하여 그 발현 양상을 검증하였다. 결 과: 자궁내막증에 이환된 여성의 자궁내막조직에서 대조군에 비하여 높게 발현되고 있는 것으로 나타난 유전자들은 ATP synthase H transporting F1 (ATP5B), eukaryotic translation elongation factor 1, isocitrate dehydrogenase 1 (NADP+), mitochondrial ribosomal protein L3, ATP synthase H+ trarsporting (ATP5C1), LPS induced TNF-$\alpha$ factor 등으로 세포의 에너지 생성과 대사과정 및 신호전달에 관여하는 유전자들이었다. 한편 자궁내막중 환자의 자궁내막조직에서 대조군에 비하여 낮게 발현된 유전자들은 insulin like growth factor II associated protein, EGF-containing fibulin-like EMP1, matrix Gla protein, TGF beta-induced, TGF beta receptor 1(activin A receptor type II-like kinase), cystallin alpha B, fibulin 5, tissue inhibitor of metalloproteinase 3, collage type XII, alpha 1, tissue inhibitor of metalloproteinase 1, decorin 등으로 세포외기질의 구성 및 기능에 관련이 있었다. 결 론: 이상의 DNA mirroarry 및 RT-PCR을 통해 얻어진 결과에서 자궁내막증의 자궁내막조직에서 대조군에 비하여 유전자들의 발현에 차이가 있음을 확인하였다.

• 한국작물학회지
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• 제63권1호
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• pp.25-34
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• 2018

휴면성이 다른 백중밀, 금강밀, 우리밀 후숙종자의 ABA 농도에 따른 발아 및 배아에서의 단백질체 발현 특성을 조사한 결과는 다음과 같다. 1. 백중밀, 금강밀, 우리밀 등 3품종의 0, 10, 30 및 $50{\mu}M$ ABA에서의 평균 발아지수와 발아율은 각각 0.95와 98% 이상으로 통계적으로 유의한 차이가 없었다. 유아와 유근의 생장은 $0{\mu}M$ ABA보다 10, 30 및 $50{\mu}M$ ABA에서 생장이 크게 억제되었는데, ABA 농도가 높을수록 생장이 더 억제되었다. 2. 3품종 배아의 평균 ABA 함량은 $0{\mu}M$ ABA와 $50{\mu}M$ ABA에서 각각 0.78 ng/mg과 269.04 ng/mg으로서 농도에 따른 ABA 함량의 차이가 컸다. 3. $0{\mu}M$ ABA 처리구에 비하여 $50{\mu}M$ ABA 처리구에서 발현양이 증가한 단백질 spot (S1, S3, S4, S6, S15, S16, S17)은 7개였으며, 감소한 단백질 spot (S2, S5, S9, S10, S11, S12, S13, S14, S18)은 8개였고, 증가와 감소가 동시에 이루어진 단백질 spot (S2)은 1개였다. $50{\mu}M$ ABA에서만 검출된 단백질 spot (S7, S8)은 2개였다. 4. 각 단백질 spot의 $0{\mu}M$ ABA 처리구 양에 대한 $50{\mu}M$ ABA 처리구 양의 평균 배수 값(fold 값)이 1.5배 이상으로 증가한 단백질 spot은 S1 (globulin-3A), S6 (globulin-1 S allele), S16 (globulin-1 S allele), S17 (globulin-1 S allele) 등으로 모두 globulin류 단백질이었다. 또한 $50{\mu}M$ ABA에서만 확인된 단백질 spot인 S7 (globulin 3)과 S8 (globulin-1 S allele)도 globulin 단백질이었다. 5. 각 단백질 spot의 $0{\mu}M$ ABA 처리구 양에 대한 $50{\mu}M$ ABA 처리구 양의 평균 배수 값(fold 값)이 0.7 이하로 감소한 단백질 spot은 S10 (glutamine sysnthetase cytosolic isozyme), S12 (S-adenosylmethionine synthetase 2), S14 (isocitrate dehydrogenase NADP)이었다. 이상의 결과는 ABA에 의한 밀 유묘의 유아와 유근의 생장억제는 배아에서의 ABA 농도 증가, 그리고 이에 따른 배아의 glutamine 합성에 관여하는 다양한 효소의 발현 감소 및 메틸기공여물질의 감소와 이에 따른 메틸기 전이활성의 감소 등이 관여하고 있음을 의미한다. 한편 배아에서의 ABA에 의한 globulin 단백질의 증가는 배아 특이적 globulin의 일시적 합성 증가와 globulin 분해 효소의 활성 억제 등이 복합적으로 관여한 결과로 생각된다.

• Korean Chemical Engineering Research
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• 제55권3호
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• pp.363-368
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• 2017

메탄올자화균이란 일탄소 화합물인 메탄올을 주탄소원 및 에너지원으로 이용할 수 있는 미생물을 말한다. Methylobacterium extorquens AM1은 serine cycle을 탄소대사경로로 이용하는 메탄올자화균 중에서도 가장 많이 연구가 진행된 균주이다. M. extorquens AM1의 poly 3-hydroxybutyrate (PHB) cycle은 EMCP (ethylmalonyl-CoA pathway), glyoxylate regeneration cycle, TCA cycle과 연결되어 있으며 EMCP 유래 유기산 또는 TCA 유기산을 생산하기 위해서는 PHB cycle로 흐르는 carbon flux의 차단이 필요하다. 이를 위해서 PHB 합성과 acetyl-CoA flux의 조절유전자로 알려져 있는 PhaR 유전자를 markerless gene deletion 방법을 이용해서 M. extorquens AM1에서 knockout했다. 결과적으로, knockout 균주인 ${\Delta}phaR$에서 야생종 대비 확연히 PHB granule이 줄어든 것이 확인되었다. Lag phase가 약 12 h 늦어졌지만, ${\Delta}phaR$은 야생종과 비슷한 세포성장과 메탄올소비 경향을 보임을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권4호
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• pp.548-555
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• 2018

Objective: This experiment was conducted to investigate whether asparagine (Asn) could improve liver energy status in weaning pigs when challenged with lipopolysaccharide. Methods: Forty-eight weaned pigs ($Duroc{\times}Large\;White{\times}Landrace$, $8.12{\pm}0.56kg$) were assigned to four treatments: i) CTRL, piglets received a control diet and injected with sterile 0.9% NaCl solution; ii) lipopolysaccharide challenged control (LPSCC), piglets received the same control diet and injected with Escherichia coli LPS; iii) lipopolysaccharide (LPS)+0.5% Asn, piglets received a 0.5% Asn diet and injected with LPS; and iv) LPS+1.0% Asn, piglets received a 1.0% Asn diet and injected with LPS. All piglets were fed the experimental diets for 19 d. On d 20, the pigs were injected intraperitoneally with Escherichia coli LPS at $100{\mu}g/kg$ body weights or the same volume of 0.9% NaCl solution based on the assigned treatments. Then the pigs were slaughtered at 4 h and 24 h after LPS or saline injection, and the liver samples were collected. Results: At 24 h after LPS challenge, dietary supplementation with 0.5% Asn increased ATP concentration (quadratic, p<0.05), and had a tendency to increase adenylate energy charges and reduce AMP/ATP ratio (quadratic, p<0.1) in liver. In addition, Asn increased the liver mRNA expression of pyruvate kinase, pyruvate dehydrogenase, citrate synthase, and isocitrate dehydrogenase ${\beta}$ (linear, p<0.05; quadratic, p<0.05), and had a tendency to increase the mRNA expression of hexokinase 2 (linear, p<0.1). Moreover, Asn increased liver phosphorylated AMP-activated protein kinase (pAMPK)/total AMP-activated protein kinase (tAMPK) ratio (linear, p<0.05; quadratic, p<0.05). However, at 4 h after LPS challenge, Asn supplementation had no effect on these parameters. Conclusion: The present study indicated that Asn could improve the energy metabolism in injured liver at the late stage of LPS challenge.