• 제목/요약/키워드: L-cysteine

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SPME/GC-MS를 이용한 혈액중의 호모시스테인, 메치오닌 및 시스테인의 분석 (Determination of Homocysteine, Methionine, Cysteine in Human Plasma with SPME/GC-MS)

  • 유은아;김정수;명승운
    • 분석과학
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    • 제13권6호
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    • pp.736-742
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    • 2000
  • SPME/GC-MS 방법을 이용하여 호모시스테인, 메치오닌 및 시스테인의 혈액중의 함량을 측정하고 관상동맥질환과 호모시스테인의 함량과의 관계를 조사하였다. 혈액중의 호모시스테인, 메치오닌 및 시스테인은 관상동맥질환의 위해성에 대한 생체표식인자로 알려지고 있다. 관상동맥질환자들에 대한 이들의 함량분포는 호모시스테인의 경우 $18.47-33.38{\mu}mol/L$,메치오닌피 경우는 $30.16-55.72{\mu}mol/L$ 그리고 시스테인의 경우는 $183.16-387.32{\mu}mol/L$ 범위의 함량을 보였다. 이 방법은 시간과 노력이 절약되고 또한 용매의 제한 사용으로 훨씬 환경 친화적인 방법임을 시사하였다.

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Studies on Thermostable Tryptophanase from a Symbiotic Thermophile

  • Chung, Yong-Joon;Beppu, Teruhiko
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 1986년도 추계학술대회
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    • pp.515.1-515
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    • 1986
  • Thermostable tryptophanase was extracted from a thermophilie bacterium, strain T which was absolutely symbiotic with strain 5. The enzyme was purified 14.7 fold with 5.8% yield by chromatographies using ion exchange, gel filtration, and hydrophobic interaction columns, followed by high performance liquid chromatography on hydroxyapatite column. The purified enzyme has a molecular weight of approximately 210,000 estimated by gel filtration column chromatography, and the molecular weight of subunit was determined by SDS polyacrylamide gel electrophoresis to be 46,000, which indicates that the native enzyme is made of four homologous subunits. The tryptophanase was stable at 65o0 and the optimum temperature for the enzyme activity for 20 min reaction was 70$^{\circ}C$. The purified enzyme activity for 20 min ieaction was 70$^{\circ}C$. The purified enzyme catalyzed the degradation of L-tryptophan into indole, pyruvate and ammonia in the presence of pyridoxal phosphate. 5-Hydroxy-Ltryptophan, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-L-cysteine, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-Lcysteine, and L-serine were also used as substrates to form pyruvate. The amino acid composition of the tryptophanase was determined, and found to contain a high percentage of hydrophobic amino acids, especially in the proline content, which was much higher than that of Escherichia coli tryptophanase. In addition, the 35N-terminal amino acid sequence of the tryptophanase was completely different from that of E. coli tryptophanase.

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Bifidobacterium breve의 성장 특성과 생존력에 미치는 배양조건의 영향 (Effect of Culture Conditions on the Growth Characteristics and Survival of Bifidobacterium breve)

  • 박희경;허태련
    • 한국식품과학회지
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    • 제28권3호
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    • pp.451-457
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    • 1996
  • Bifidobacterium breve의 최적 배양 조건과 안정성을 증진시킬 수 있는 배양조건을 조사하기 위하여 pH와 L-cysteine HCI의 첨가효과를 조사하였다. 최대 균체수를 얻기위한 pH 조건은 pH $6.0{\sim}6.5$가 최적 배양 조검임을 알 수 있었다. 저장시의 최적 배양 조건은 $5.5{\sim}6.0$에서 가장 우수하였다. pH $5.5{\sim}6.0$에서 배양한 균주를 $4^{\circ}C$에서 저장시 25일 이후에 $2.4{\times}10^6/ml$$1.4{\times}10^6/ml$$10^6/ml$ 이상을 유지하여 저장시 안정성이 우수한 반면 pH 7.0에서 배양한 균주는 $2.4{\times}10^6/ml$로 저장시 생존력이 상당히 낮았으며 pH 5.5와 6.0에서 배양한 균주의 ${\beta}$-galactosidase 활력은 25일 저장 이후에도 $78{\sim}85%$를 유지한 반면 pH 7.0에서 배양한 균주의 ${\beta}$-galactosidase 활력은 급격히 감소하였다. L-cysteine HCI 첨가 효과는 0.05% 이상 첨가시 증식에 유리한 환경을 제공하였으며 과산화수소에 대한 내성은 $0.05{\sim}0.10%$ L-cysteine HCI을 첨가한 배이에서 증식한 B. breve가 우수하였다. Osmoprotectant를 첨가하여 배양시 증식과 동결건조시 균주의 안정성에 대한 효과를 조사해 본 결과 2mM betaine이나 2mM trehalose를 첨가시 증식에도 효과가 있었으며 osmoprotectant 역할을 함을 알 수 있었다. 또 한 동결건조에서도 betaine이나 trehalose를 첨가하여 배양된 B. breve는 거의 손상을 받지 않음을 알 수 있었다.

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Isolation and characterization of a cDNA encoding a mammalian cathepsin L-like cysteine proteinase from Acanthmoeba healui

  • Hong, Yeon-Chul;Hwang, Mi-Yul;Yun, Ho-Cheol;Yu, Hak-Sun;Kong, Hyun-Hee;Yong, Tai-Soon;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • 제40권1호
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    • pp.17-24
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    • 2002
  • We have cloned a cDNA encoding a cysteine proteinase of the Acanthamoeba healui OC-3A strain isolated from the brain of a granulomatous amoebic encephalitis patient. A DNA probe for an A. healui cDNA library screening was amplified by PCR using degenerate oligonucleotide primers designed on the basis of conserved amino acids franking the active sites of cysteine and asparagine residues that are conserved in the eukaryotic cysteine proteinases. Cysteine proteinase gene of A. healui (AhCPI) was composed of 330 amino acids with signal sequence, a proposed pro-domain and a predicted active site made up of the catalytic residues, $Cys^{25},{\;}His^{159},{\;}and{\;}Asn^{175}$. Deduced amino acid sequence analysis indicates that AhCPI belong to ERFNIN subfamily of C 1 peptidases. By Northern blot analysis. no direct correlation was observed between AhCPI mRNA expression and virulence of Acanthamoeba, but the gene was expressed at higher level in amoebae isolated from soil than amoeba from clinical samples. These findings raise the possibility that AhCPI protein may play a role in protein metabolism and digestion of phagocytosed bacteria or host tissue debris rather than in invasion of amoebae into host tissue.

Dichloro, Alanine and S-Methylcysteine Cobalt (III) Complexes of Ethylenediamine-N,N$^\prime$-di-$\alpha$-isobutyric Acid

  • Jun, Moo-Jim;Park, Chang-Woo;Park, Youn-Bong;Cheon, Jin-Woo;Choi, Sung-Rack
    • Bulletin of the Korean Chemical Society
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    • 제11권4호
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    • pp.354-357
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    • 1990
  • Synthesis of dichloro cobalt (Ⅲ) complexes of a flexible $N_2O_2-type$ tetradentate ligand, ethylenediamene-N,N'-di-${\alpha}$-isobutyric acid (eddib), has yielded two geometrical isomers, s-cis-$(Co(eddib)Cl_2)- and uns-cis-(Co(eddib)Cl_2)-.$ A series of substitution reactions, $(Co(eddib)Cl_2)^- {\to} (CO(eddib)Cl H_2O) {\to} (Co(eddib)CO_3)^- {\to} (Co(eddib(H_2O)_2)^+$ have been run for each of the two geometrical isomers. The reaction between the s-cis-(Co(eddib)Cl_2)^-$ complex and L-alanine (L-als) or S-methyl-L-cysteine (L-mcy) gave the meridional s-cis-[Co(eddib)(aa)) (aa = L-ala or L-mcy) complex. The S-methyl-L-cysteine was found to coordinate to cobalt (Ⅲ) ion via the nitrogen and oxygen donor atoms.

Determination of L-FMAUS, a new L-FMAU derivative, in rat plasma and urine by high-performance liquid chromatography

  • Chung, Hye-Jin;Kim, Yu-Chul;Koo, Chang-Hui;Lee, Myung-Gull
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.277.1-277.1
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    • 2003
  • A high-performance liquid chromatographic method using the liquid extraction procedure was developed for the determination of L -FMAUS. a new L -FMAU derivative, in rat plasma and urine using 3-aminophenyl sulfone as an internal standard. A 100-${\mu}\ell$ aliquot of distilled water containing the L -cysteine (100 mg/$m\ell$) was added to a 100-${\mu}\ell$ aliquot of biological sample. L-Cysteine was employed to protect binding between 5'-thiol of l and protein in the biological sample. (omitted)

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품종별 마늘의 품질 특성 및 흑마늘 숙성 중 품질 변화 (Physicochemical characteristics of domestic garlic by varieties and their quality changes during aging of black garlic)

  • 유승희;차건수;김동규;강민정
    • 한국식품저장유통학회지
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    • 제30권3호
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    • pp.446-458
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    • 2023
  • 우리나라 대표적인 남도, 대서, 홍산과 의성 마늘의 품질특성을 비교하고 각각의 마늘을 흑마늘로 숙성하면서 유효물질의 변화를 조사하였다. 흑마늘은 60-80℃의 범위에서 15일간 숙성하면서 5일 단위로 시료를 취하였다. 총페놀화합물은 생마늘에서 85.2-109.7 mg GAE/100 g이던 것이 숙성기간이 증가함에 따라 증가하였고, 생마늘과 제조된 흑마늘 모두 홍산마늘이 가장 높았다. 알린 함량은 생마늘에서 848.3-1,087.5 mg/100 g이던 것이 숙성 5일에 7.3-20.6 mg/100 g으로 유의하게 감소한 후 더 감소하였다. 생마늘의 GSAC(γ-glutamyl-S-allyl-cysteine)와 GS1PC (γ-glutamyl-S-1-Propenyl-l-Cysteine) 함량은 202.1-541.0 mg/100 g, 311.1-474.3 mg/100 g이었으며 숙성 동안 점차 감소하였다. 한편, SAC(S-allyl-cysteine) 및 S1PC (S-1-Propenyl-l-Cysteine) 함량은 4종 모두 생마늘일 때 매우 낮았으나 숙성 5일 차에 크게 증가한 후 감소하는 경향을 보였다. 숙성 5일째 SAC 함량은 남도마늘이 208.0 mg/100 g으로 다른 시료에 비해 유의적으로 높았고, S1PC는 의성마늘이 66.2 mg/100 g으로 유의적으로 높았다. 이는 생마늘에서 SAC 및 S1PC의 전구물질인 GSAC와 GS1PC가 높았던 시료가 흑마늘 숙성 후에도 높은 경향이었다. 따라서 마늘은 품종에 따라 유효물질의 함량에 차이가 있고 숙성 후의 주요 화합물의 변화에도 유의적인 차이가 있을 것으로 추측되어 향후 기능성 식품 연구의 기초자료로 활용될 것으로 판단된다.

마늘의 조리방법에 따른 S-Allyl-L-Cysteine 함량 변화 (Effect of Cooking Methods on S-Allyl-L-Cysteine Content of Garlic)

  • 이상훈;정윤숙;황경아;송진;노건민;황인국
    • 한국식품영양과학회지
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    • 제45권8호
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    • pp.1221-1226
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    • 2016
  • 본 연구에서는 마늘의 기능성분인 S-allyl-L-cysteine(SAC) 분석법을 검증하고 조리방법에 따른 마늘의 SAC 함량을 분석하였다. SAC 분석법의 유효성을 검증하기 위하여 특이성, 직선성, 검출한계, 정량한계, 정밀성 및 정확성을 평가하였다. SAC 표준물질과 마늘 추출물을 유도체화시킨 후 fluorescence detector로 분석한 결과 동일한 머무름 시간을 갖는 단일 피크를 확인하였다. 직선성은 상관계수 값이 0.9999였으며, 검출한계는 $0.15{\mu}g/mL$, 정량한계는 $0.47{\mu}g/mL$, 정밀성의 상대표준편차는 5% 이하, 정확성인 회수율은 95% 이상으로 우수하였다. 생마늘의 SAC 함량은 2.77 mg/g이었으며, 데치기와 전자레인지 처리는 조리 수준에 따라 SAC 함량에 차이를 보이지 않았다. 삶기와 고압멸균처리 시 조리시간이 증가함에 따라 SAC 함량은 증가하였고, 특히 $121^{\circ}C$에서 60분 고온고압처리 할 경우 50.24 mg/g으로 증가하였다.

Amplification of Glutathione Production in E. coli Cells Using Recombinant DNA Techniques

  • Nam, Yong-Suk;Park, Young-In;Lee, Se-Yong
    • Journal of Microbiology and Biotechnology
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    • 제1권3호
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    • pp.157-162
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    • 1991
  • Conditions for glutathione production in E. coli cells which possess pGH501 (2 gshI+gshII) were studied. In terms of ATP supply for the glutathione synthesis, two different systems have been constructed and compared. When the acetate kinase reaction of E. coli was used for ATP generation, 20 mM of L-cysteine was completely converted to glutathione by toluene-treated E. coli cells (100 mg/ml) harboring pGH501 within 2 h at $37^{\circ}C$. However, considering the economical aspects, the glycolytic pathway of yeast was chosen as a better system for ATP generation. The optimal concentrations of reactants for glutathione production were determined to be as follows; 80 mM L-glutamate, 20 mM L-cysteine, 20 mM glycine, 20 mM $MgCl_2$, 50 mM potassium phosphate buffer (pH 7.5), 400 mM glucose, polyoxyethylene stearylamine ($5\;\mul/ml$), toluene-treated E. coli HB101/pGH501 (100 mg/ml), and dried yeast cells (400 mg/ml). The conversion ratio of L-cysteine to glutathione was 80% (about 5 mg/ml) under optimal condition within 6 h at $37^{\circ}C$.

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Chemical Modification of Cysteine Residues in Hafnia alvei Aspartase by NEM and DTNB

  • Shim, Joon-Bum;Kim, Jung-Sung;Yoon, Moon-Young
    • BMB Reports
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    • 제30권2호
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    • pp.113-118
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    • 1997
  • Aspartase from Hafnia alvei was inactivated by N-ethylmaleimide (NEM) and 5,5' -Dithiobis-(2-znitrobenzoic acid) (DTNB) following pseudo-first order kinetics. Their apparent reaction orders were 0.83 and 0.50 for NEM and DTNB modifications, respectively, indicating that inactivation was due to a sulfhydryl group in the active site of aspartase and participation of the sulfhydryl group in an essential step in the catalytic reaction. When aspartase was modified by DTNB, the enzyme activity was restored by dithiothreitol treatment, indicating that cysteine residuetsl islarel possibly at or near the active site. The pH-dependence of the inactivation rate by NEM suggested that an amino acid residue having pK value of 8.3 was involved in the inactivation. When aspartase was incubated with NEM and L-aspartate together, L-aspartate markedly protected the enzyme from inactivation by NEM, but the other reagents used did not.

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