• 제목/요약/키워드: Gln3

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Bacillus amyloliquefaciens HC188이 생산하는 혈전분해 효소의 정제 및 특성 (Purification and Characterization of a Fibrinolytic Enzyme Produced by Bacillus amyloliquefaciens HC188)

  • 신소희;홍성욱;정건섭
    • 한국미생물·생명공학회지
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    • 제41권1호
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    • pp.33-43
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    • 2013
  • 식품으로서 안전하게 섭취하여 혈전증을 사전에 예방하거나 개선할 수 있도록 하기 위하여, 전통적 방법으로 제조한 청국장으로부터 미생물을 분리하여 혈전분해 효소활성이 우수한 미생물을 선발 동정한 결과, Bacillus amyloliquefaciens HC188이라 명명하였다. 선발미생물이 생산하는 혈전분해효소를 분리 및 정제한 결과, 50.7배의 정제도와 5.5%의 수율을 나타내었고, 혈전분해 효소단백질의 분자량은 22.3 kDa이었으며, N-terminal 아미노산 서열은 Ala-Gln-Ser-Val-Pro-Tyr-Gly-Val-Ser-Gln-Ile-Lys-Ala-Pro-Ala로 분석되었다. 효소의 최적반응 pH와 온도는 pH 8.0과 $40^{\circ}C$로 나타났으며, pH 6.0-8.0과 $20-40^{\circ}C$ 사이에서 효소가 비교적 안정하였다. 금속이온에 대한 영향은 2 mM과 5 mM 농도의 $CoCl_2$$CaCl_2$의 금속이온이 존재할 때 효소활성이 증가하였으며, inhibitor로서 EDTA와 PMSF에 의해 효소활성이 저해되므로 청국장에서 분리한 B. amyloliquefaciens HC188가 생성한 혈전분해 효소는 metallo-serine protease로 사료되었다.

베타 2-아드레날린 수용체의 유전자 변이형이 비만도에 미치는 영향 (Effects of ${\beta}2-Adrenergic$ Receptor Polymorphism on Obesity)

  • 김길수;오현희;최선미;양현성;배정환;윤유식
    • 한방비만학회지
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    • 제3권1호
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    • pp.7-16
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    • 2003
  • Objectives : The lipolytic effects of catecholamines in adipose tissue are mediated by members of adrenergic receptors. This study was conducted to examine the effects of ${\beta}2-AR$ Gln27Glu (Q27E) polymorphism on obesity indices and risk among obesity clinic patients. Methods : 532 subjects, 38 men and 494 women, who attended a weight loss program in a local obesity clinic were analyzed. Height, weight, BMI, WHR and obesity index were measured or calculated. Body composition was measured by bio-impedance analysis. Genotype of ${\beta}2-AR$ polymorphism in codon 27 was analyzed by PCR-RFLP method. Serum concentrations of fasting glucose, total and HDL cholesterol, and triglyceride were determined by autobiochemical analyzer. Results: The Genotype distributions of ${\beta}2-AR$ gene were QQ type 81.3%, QE type 17.9% and EE type 8%. Therefore, the frequency of E allele of ${\beta}2-AR$ gene was 0.170 in the total subjects. The frequency of the ${\beta}2-AR$ variant genotype(QE+EE) was significantly higher in obese group($BMI{\geqq}25$) compared with non-obese group(p=0.027). Weight was significantly higher in variant(QE+EE) type compared with normal(QQ) type in total subjects(p=0.001), male(p=0.022) and female(p=0.013). BMI, obesity index and WHR were also significantly higher in QE+EE type. Body fat man was significantly higher in QE+EE type in total subjects(p=0.005) and female(p=0.027). When forward stepwise regression analysis was used to create a model of risk predictors of obesity($BMI{\geqq}25$), QE+EE type of ${\beta}2-AR$ gene was found to be a significant risk factor for obesity (p=0.042, ORs 1.597). Conclusions: QE+EE genotype of ${\beta}2-AR$ was associated with increased obesity indices and might be a significant risk factor for obesity.

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$\beta_2$ 교감신경 수용체 다형성이 아토피 및 혈청 IgE에 미치는 영향 (Association Between $\beta_2$ Adrenoceptor Polymorphisms and Atopy/serum IgE in Asthmatic Patients)

  • 이신형;심재정;강용구;정혜철;김경규;권영환;김제형;이승룡;이소라;이상엽;조재연;인광호;유세화;강경호
    • Tuberculosis and Respiratory Diseases
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    • 제46권6호
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    • pp.826-835
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    • 1999
  • 연구배경 : $\beta_2$ 교감신경 수용체의 다형성이 기관지 천식의 표현형에 영향을 미친다고 알려져 있다. 이에 저자들은 먼저 기관지 천식 환자와 정상인에서의 $\beta_2$ 교감신경 수용체의 다형성의 빈도에 차이가 있는가를 알아보고, 또한 기관지 천식 환자에서 아토피의 유무 및 혈청 총 IgE의 증가 여부와 $\beta_2$ 교감신경 수용체의 다형성이 관계가 있는지 알아보고자 본 연구를 시행하였다. 방 법 : 기관지 천식 환자 109명과 정상인 42명에 대하여 혈청 총 IgE를 측정하고, 항원 특이 IgE 검사 및 피부 단자 검사를 실시하였고, mutated allele specific amplification 법으로 $\beta_2$ 교감신경 수용체와 다형성을 검색하였다. 결 과 : $\beta_2$ 교감신경 수용체의 다형성을 조사한 결과 16번 아미노산 위치에서는 Arg 야생형 및 Arg/Gly 이형접합체 변이형, Gly 동형 접합체 변이형이, 그리고 27번은 Gln 야생형, Gln/Glu 이형 접합체 변이형, Glu 동형 접합체 변이형이 관찰되었다. 34번 아미노산의 경우는 Val 야생형과 Val/Met 이형 접합체 변이형이, 164번은 Thr 야생형만 있었다. 1) 정상인과 기관지 천식 환자에서 $\beta_2$ 교감신경 수용체의 16, 27, 34번 아미노산 위치에 있어서 다형성의 빈도는 통계학적으로 차이가 없었다(p>0.05)(Table 3). 2) 기관지 천식환자에서 $\beta_2$ 교감신경 수용체 다형성의 빈도와 아토피의 존재 유무는 16, 27, 34 번 아미노산에서 모두 통계학적으로 관계가 없었다(p>0.05) (Table 4). 3) 기관지 천식 환자 중 혈청 총 IgE가 증가된 군과 정상 군에서 $\beta_2$ 교감신경 신경 수용체 다형성과 혈청 총 IgE의 증가 여부의 비교는 16, 27, 34번 아미노산 모두 통계학적인 관련이 없었다(p>0.05) (Table 5). 결 론 : 결론적으로 기관지 천식 환자와 정상인 사이의 $\beta_2$ 교감신경 수용체의 다형성은 16, 27, 34번 아미노산 위치에서 차이가 없었고, 기관지 천식환자에서 아토피의 유무 및 혈청 총 IgE의 증가 여부와 $\beta_2$ 교감신경 수용체의 다형성의 빈도는 통계적으로 유의한 관계가 없었다.

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Computational Analysis of Apolipophorin-III in Hyphantria cunea

  • Chandrasekar R.;Dhanalakshmi R.;Krishnan M.;Kim H. J.;Jeong H. C.;Seo S. J.
    • International Journal of Industrial Entomology and Biomaterials
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    • 제10권1호
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    • pp.25-33
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    • 2005
  • Recently a cDNA clone of apoLp-III from Hyphantria cunea was isolated and subjected to computational analysis to compare with other available sequences. Multiple sequence alignments were carried out using the amino acid sequences of apoLp-III from six insects. It was found that the H. cunea apoLp-III has relatively high sequence identities to Spodoptera litura ($69.5\%$), Manduca sexta ($66.8\%$), Galleria mellonella ($65.1\%$), Bombyx mori N4 ($54.3\%$) but less identity to Locusta migratoria ($18.3\%$). The amino acid composition was compared with other insects using EXPASY tools; it shows that alanine (Ala), glutamine (Gln), leucine (Leu) and lysine (Lys) are the major amino acid components of apoLp-III in H. cunea as well as other lepidopterans. Homology modeling performed using PSI-BLAST (PDB template M. sexta) reveals that the apoLp-III molecules consist of five, long amphipathic alpha helical bundles with short loops connecting the helices and shows homology with other insects. Phylogenetic analysis shows that the orthopteran apoLp-III represented by locust was most distantly related to the lepidopteran insects.

Receptor-oriented Pharmacophore-based in silico Screening of Human Catechol O-Methyltransferase for the Design of Antiparkinsonian Drug

  • Lee, Jee-Young;Baek, Sun-Hee;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • 제28권3호
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    • pp.379-385
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    • 2007
  • Receptor-oriented pharmacophore-based in silico screening is a powerful tool for rapidly screening large number of compounds for interactions with a given protein. Inhibition of the enzyme catechol-Omethyltransferase (COMT) offers a novel possibility for treating Parkinson's disease. Bisubstrate inhibitors of COMT containing the adenine of S-adenosylmethionine (SAM) and a catechol moiety are a new class of potent and selective inhibitor. In the present study, we used receptor-oriented pharmacophore-based in silico screening to examine the interactions between the active site of human COMT and bisubstrate inhibitors. We generated 20 pharmacophore maps, of which 4 maps reproduced the docking model of hCOMT and a bisubstrate inhibitor. Only one of these four, pharmacophore map I, effectively described the common features of a series of bisubstrate inhibitors. Pharmacophore map I consisted of one hydrogen bond acceptor (to Mg2+), three hydrogen bond donors (to Glu199, Glu90, and Gln120), and one hydrophobic feature (an active site region surrounded by several aromatic and hydrophobic residues). This map represented the most essential pharmacophore for explaining interactions between hCOMT and a bisubstrate inhibitor. These results revealed a pharmacophore that should help in the development of new drugs for treating Parkinson's disease.

Influence of Amino Acidic Additives on Properties of EPDM-g-MAH/ZnO Composites

  • Choi, Sung-Seen;Kim, Yeowool;Chung, Yu Yeon;Bae, Jong Woo;Kim, Jung-Soo
    • Elastomers and Composites
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    • 제51권3호
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    • pp.175-180
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    • 2016
  • Influence of amino acidic chemical on properties of maleic anhydride-grafted ethylene-propylene-diene terpolymer/zinc oxide (EPDM-g-MAH/ZnO) composites was investigated. 4-Aminosalicylic acid (ASA), 4-amino-2-methoxybenzoic acid (AMBA), 12-aminolauric acid (ALA), and glutamine (Gln) were employed as the amino acidic chemicals. Though small quantity (0.5 phr) of the amino acidic chemical was added to the EPDM-g-MAH/ZnO composite, the properties were notably changed. By adding the amino acidic chemical, the percent crystallinity and apparent crosslink density were reduced. Order of the percent crystallinity was related to that of the $pK_a$ values of amino acidic chemicals. By adding the amino acidic chemical, the basic tensile properties were on the whole improved. The experimental results were explained by the $pK_a$ values of amino acidic chemicals, change of zinc ionomer formation, and interactions between the additive and EPDM-g-MAH chain.

바이오패닝에 의한 Pb2+ 친화성 펩타이드 서열의 탐색 (Screening of Peptide Sequences Cognitive of Pb2+ by Biopanning)

  • ;홍순호;최우석;유익근
    • KSBB Journal
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    • 제28권3호
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    • pp.185-190
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    • 2013
  • For the selection of peptide specifically binding to $Pb^{2+}$, the biopanning with the commercially available Ph.D.-7 phage displayed heptapeptide library was carried out against $Pb^{2+}$ immobilized on a metal-chelating IDA (iminodiacetic acid) resin. After four rounds of screening against $Pb^{2+}$-IDA including negative selections against charged bead with metal ions other than $Pb^{2+}$ and uncharged bead, several candidate lead-binding phage peptides were initially determined based on the order of frequency from the screened phage clones. Of the selected phage peptide sequences, the peptide of the highest frequency, CysSerIleArgThrLeuHisGlnCys (CSIRTLHQC) also exhibited the strongest affinity for $Pb^{2+}$ in binding assays for individual phage clones. However, there was not a significant difference in $Pb^{2+}$ affinity between selected peptides when using synthetic heptapeptides corresponding to the displayed peptide sequences of phage clones.

Mechanism Studies of Substituted Triazol-1-yl-pyrimidine Derivatives Inhibition on Mycobacterium tuberculosis Acetohydroxyacid Synthase

  • Chien, Pham Ngoc;Jung, In-Pil;Reddy, Katta Venugopal;Yoon, Moon-Young
    • Bulletin of the Korean Chemical Society
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    • 제33권12호
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    • pp.4074-4078
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    • 2012
  • The first step in the common pathway for the biosynthesis of branched chain amino acids is catalyzed by acetohydroxyacid synthase (AHAS). The AHAS is found in plants, fungi and bacteria. With an aim to identify new anti-tuberculosis drugs that inhibit branched chain amino acid biosynthesis, we screened a chemical library against Mycobacterium tuberculosis AHAS. The screening identified four compounds, AVS 2087, AVS 2093, AVS 2236, and AVS 2387 with $IC_{50}$ values of 0.28, 0.21, 3.88, and $0.25{\mu}M$, respectively. Moreover, these four compounds also showed strong inhibition against reconstituted AHAS with $IC_{50}$ values of 0.37, 0.26, 1.0, and $1.18{\mu}M$, respectively. The basic scaffold of the AVS group consists of 1-pyrimidin-2-yl-1H-[1,2,4]-triazole-3-sulfonamide. The most active compound, AVS 2387, showed the lowest total interaction energy -8.75 Kcal/mol and illustrates its binding mode by hydrogen bonding with $H_{\varepsilon}$ of Gln517 with the distance of $2.24{\AA}$.

Electron Redistribution of Clavalanate on Binding to a $\beta$-Lactamase

  • Sang-Hyun Park;Hojing Kim
    • Bulletin of the Korean Chemical Society
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    • 제14권4호
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    • pp.491-496
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    • 1993
  • A class A ${\beta}$-lactamase from Staphylococcus aureus PC1 complexed with 3R,5R-clavulanate is studied. The starting geometry for the computation is the crystal structure of the ${\beta}$-lactamase. Docking of the clavulanate to the enzyme is done exploiting the requirements of electrostatic and shape complementarity between the enzyme and clavulanate. This structure is then hydrated by water molecules and refined by energy minimization and short molecular dynamics simulation. In the energy refined structure of this complex, the carboxyl group of the clavulanate is hydrogen bonded to Lys-234, and the the carbonyl carbon atom of the clavulanate is adjacent to the $O_{\gamma}$ of Ser-70. It is found that a crystallographic water molecule initially located at the oxyanion hole, which is formed by the two -NH group of Ser-70 and Gln-237, is replaced by the carbonyl oxygen atom of the 3R,5R-clavulanate after docking and energy reginement. The crystallographic water molecules are proved to be important in ligand binding. Glu-166 residue is found to be repulsive to the binding of clavulanate, which is in agreement with experimental observation. Arg-244 residue is found to be important to the binding of clavulanate as well as to interaction with C2 side chain of the clavulanate. The electron density redistribution of the clavulanate on binding to the ${\beta}$-lactamase in studied by an ab initio quantum-mechanical calculation. A significant redistribution of electron density of the clavulanate is induced by the enzyme, toward the enzyme, toward the transition state of the enzymatic reaction.

우렁쉥이 육으로부터 Carotenoprotein의 분리.정제 및 특성 (Purification and Characterization of Carotenoprotein from the Muscle of Ascidian, Halocynthia roretzi)

  • 강옥주;변희국;전유진;김종배;김세권
    • Applied Biological Chemistry
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    • 제41권1호
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    • pp.31-41
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    • 1998
  • 우렁쉥이의 육에 존재하는 carotenoprotein을 추출하여 이온교환수지 및 겔크로마토그래피를 이용하여 정제하였으며, 정제된 carotenoprotein의 분자량, pH 안정성, 열안정성, 변성제의 영향, 아미노산 조성 및 지방산 조성을 분석하였다. Carotenoprotein의 정제도는 5배, 수율은 2.9%였으며, 1 mg당 $0.15\;{\mu}g$ carotenoid가 함유되어 있었다. Carotenoprotein의 최대 흡수파장은 UV/Vis 스펙트럼에서 463 nm 및 439 nm였으며, 분자량은 64.4 kDa으로 단일 polypeptide로 이루어져 있었다. Carotenoprotein은 pH 7.0부근의 중성영역에서 안정하였고, 열안정성은$65^{\circ}C$에서 2시간 이내에 90%이상 유지하였으며, SDS 변성제에 의해 5 nm 단파장 영역으로 이동하였다. Carotenoprotein의 아미노산 조성은 Gly, hsn, Gln 및 Ser의 함량이 전체 아미노산의 35.3%로 전체적으로 산성의 성질을 보였으며, 구성지방산 조성의 함량은 monoene산이 45.2%로 가장 많았고, 다음으로 포화지방산 및 polyene산이 각각 38.1% 및 11.7%였다.

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