• 제목/요약/키워드: active site

검색결과 1,209건 처리시간 0.033초

Effect of His192 Mutation on the Activity of Alginate Lyase A1-III from Sphingomonas Species A1

  • Yoon, Hye-Jin;Choi, Yong-Jin;Osamu Miyake;Wataru Hashimoto;Kousaku Murata;Bunzo Mikami
    • Journal of Microbiology and Biotechnology
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    • 제11권1호
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    • pp.118-123
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    • 2001
  • The alginate lyase A1-III gene of Sphingomonas species A1 is composed of 1,077 nucleotides, encoding a protein (359 amino acids) with a molecular mass of 40,322 Da. Recombinant A1-III expressed in Escherichia coli exhibited the same full enzymatic activity as native A1-III. In order to identify the critical residue for activity, a site-directed mutation was introduced into the A1-III gene (H192A, His192->Ala). Recombinant A1-III (H192A) exhibited a significant decrease in enzyme activity (one-thirty thousandth of that of A1-III), without any conformational change, as detected by the CD spectra in the far UV region. Also, the chemical modification of wild-type A1-III with methyl 4-nitro benzene sulfonate resulted in a 40% decrease from the initial activity, whereas the same modification of A1-III (H192A) produced no change in the activity. The role of His192 on the catalytic process was also explored based on a model of A1-III docked with mannuronic acid into the active site.

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The active site and substrate binding mode of 1-aminocyclopropane-1- carboxylate oxidase of Fuji apple (Malus domesticus L.) determined by site directed mutagenesis and comparative modeling studies

  • Ahrim Yoo;Seo, Young-Sam;Sung, Soon-Kee;Yang, Dae-Ryook;Kim, Woo-Tae-K;Lee, Weontae
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.70-70
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    • 2003
  • Active sites and substrate bindings of 1-aminoxyclopropane-1-carboxylate oxidase (MD-ACO1) catalyzing the oxidative conversion of ACC to ethylene have been determined based on site-directed mutagenesis and comparative modeling methods. Molecular modeling based on the crystal structure of Isopenicillin N synthase (IPNS) provided MD-ACO1 structure. MD-ACO1 protein folds into a compact jelly roll shape, consisting of 9 ${\alpha}$-helices, 10 ${\beta}$-strands and several long loops. The MD-ACO1/ACC/Fe(II)/Ascorbate complex conformation was determined from automated docking program, AUTODOCK. The MD-ACO1/Fell complex model was consistent with well known binding motif information (HIS177-ASP179-HIS234). The cosubstrate, ascorbate is placed between iron binding pocket and Arg244 of MD-ACO1 enzyme, supporting the critical role of Arg244 for generating reaction product. These findings are strongly supported by previous biochemical data as well as site-directed mutagenesis data. The structure of enzyme/substrate suggests the structural mechanism for the biochemical role as well as substrate specificity of MD-ACO1 enzyme.

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Computational screening of electroactive indolequinone derivatives as high-performance active materials for aqueous redox flow batteries

  • Han, Young-Kyu;Jin, Chang-Soo
    • Current Applied Physics
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    • 제18권12호
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    • pp.1507-1512
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    • 2018
  • The development of an organic-based aqueous redox flow battery (RFB) using quinone as an electroactive material has attracted great attention recently. This is because this battery is inexpensive, produces high energy density, and is environment friendly in stationary electrical energy storage applications. Herein, we investigate the redox potentials and solubilities of indole-5,6-quinone and indole-4,7-quinone derivatives in terms of the substituent effects of functional groups using theoretical calculations. Our results indicate that full-site substituted derivatives of indolequinone are more useful as active materials compared to single-site substituted derivatives. In particular, our calculations reveal that the substitution of $-PO_3H_2$ and $-SO_3H$ functional groups with multiple polar bonds is very effective in increasing the activity of the aqueous RFB. As a strategy to overcome the limitation that the aqueous solubility is intrinsically low because they are organic molecules, we suggest the substitution of functional groups with multiple polar bonds to the backbones of active organic materials. Among 180 indolequinone derivatives, 17 candidates that meet the redox potential standards ($${\leq_-}0.2V$$ or $${\geq_-}0.9V$$) and eight candidates with solubility exceeding 2 mol/L are identified. Three indolequinone derivatives that satisfy both conditions are finally presented as promising electroactive candidates for an aqueous RFB.

국립공원내 야영장의 이용 만족도 분석 -설악산 국립공원 설악동 지구를 중심으로- (Analysis of visitor's satisfaction on camp site in national park -Focused on Sorakdong district in Mount Sorak National Park­)

  • 조태동;김미진;신병철
    • 한국환경과학회지
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    • 제14권5호
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    • pp.457-461
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    • 2005
  • The survey of user's satisfactory degree of location, plantings, facilities, operation, and management factors on national parks' camp sites was carried out. Many problems were analyzed and identified. This survey was for proposing the correct way to plan and draft for camping site after this. In addition to, the realization of environmental education through nature friendly recreation was aimed at. According to the result of the survey, the followings were suggested as necessities to improve the camp site. 1. Repair and replacement of insufficiently managed facilities should be aimed at. 2. Facilities for a handicapped person should be preferentially introduced. 3. Active management and publicity in low-demand season that practiced by developed countries should be carried out. Seasonal program for various age groups should be operated. 4. Surrounding tourist site, camp site, and trail should be organically connected by shuttle bus and other transports. A survey was carried out for the user satisfaction in the campsites of national parks. Based on this survey concerned with the location, planting, facilities, operation and management, the problems were identified and analysed to improve future design and plan of the campsites. This study aims for the realization of environmental education through nature friendly recreation. The following points are suggested as needed according to the survey result. 1. damaged facilities should be repaired or replaced. 2. introduction of the facilities for disabled should be prioritized. 3. consistent and active management and publicity work should be operated in the off-peak season. and the seasonal activity programs for the various age groups should be provided. 4. the site should be well connected with other surrounding campsites and tourist sites using the shuttle bus and other transport method.

Power Tracing을 이용한 VSC HVDC 설치위치 선정 (VSC HVDC Site Selection Using Power Tracing)

  • 오세승;장길수;문승일
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2007년도 추계학술대회 논문집 전력기술부문
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    • pp.162-164
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    • 2007
  • This paper presents a HVDC site selection algorithm to increase transfer capability using VSC HVDC system which can control active power as well as reactive power. Using normal powerflow results and simple index $k_r$ the HVDC site selection algorithm is enhanced and more tightly-coupled transmission lines are identified in a domain of generators.

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Site-specific Mutagenesis에 의한 PRD1 DNA Polymerase의 활성부위 결정 (Determination of Active Site in PRD1 DNA Polymerase by Site-specific Mutagenesis)

  • 황정원;정구홍
    • 미생물학회지
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    • 제29권4호
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    • pp.209-214
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    • 1991
  • The PRD1 DNA polymerase is a small multi-functional enzyme containing conserved amino acid sequences shared by family B DNA polymerases. Thus the PRD1 DNA polymerase provides an useful model system with which to study structure-functional relationships of DNA polymerase molecules. In order to investigate the functional and structural roles of the highly conserved amino acid sequences, we have introduced three mutations into a conserved amino acid of the PRD1 DNA polymerase. Genetic complememtation study indicated that each mutation inactivated DNA polymerase catalytic activity.

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화학수식제에 의한 Bacillus alcalophilus AX2000 유래 Xylanase의 활성에 관여하는 아미노산 잔기의 확인 (Identification of Amino Acid Residues Involved in Xylanase Activity from Bacillus alcalophilus AX2000 by Chemical Modifiers)

  • 박영서
    • 한국미생물·생명공학회지
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    • 제34권2호
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    • pp.121-128
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    • 2006
  • Bacillus alcalophilus AX2000으로부터 xylanase를 정제한 후 효소의 활성부위를 조사하기 위하여 여러 가지 화학수식제를 사용하여 효소활성의 저해도를 측정하였다. 여러 가지 화학 수식제 중에서 carbodiimide와 N-bromosuccinimide가 효소 활성을 완전히 저해시켜 glutamic acid또는 aspartic acid 잔기와 tryptophan 잔기가 효소의 활성부위에 관여하리라 추측되었다. 각각의 경우에 효소 실활은 수식제의 첨가농도에 따라 pseudo first-order kinetics 양식을 보여주었으며, carbodiimide와 N-bromosuccinimide는 각각 비경쟁적 저해와 경쟁적 저해방식을 나타내었다. 기질첨가에 의한 효소활성 보호실험을 통하여 tryptophan 잔기가 기질결합부위라 판단 되었다. 효소 실활속도의 분석에 의해 효소활성에는 2개의 glutamic acid 또는 aspartic acid 잔기와 1개의 tryptophan 잔기가 관여하는 것으로 나타났다.

MCL 알고리즘을 이용한 단백질 표면의 바인딩 영역 분석 기법 (Investigating Binding Area of Protein Surface using MCL Algorithm)

  • 정광수;유기진;정용제;류근호
    • 정보처리학회논문지D
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    • 제14D권7호
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    • pp.743-752
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    • 2007
  • 단백질은 다른 물질과의 결합하여 기능을 수행하기 때문에 활성 사이트가 유사한 단백질은 유사한 기능을 가진다. 따라서 단백질의 바인딩 영역을 식별함으로써 단백질의 기능을 추론할 수 있다. 이 논문은 MCL (Markov Cluster) 알고리즘을 이용하여 단백질의 바인딩 영역을 추출하는 새로운 방법을 제시한다. 이를 위하여 단백질의 표면 잔기 거리를 나타내는 distance matrix를 생성하고, 여기에 MCL 프로세스를 적용한다. 제시한 방법을 평가하기 위해 Catalytic Site Atlas (CSA) 데이터를 사용하였다. CSA 데이터 (94개의 단일 체인 단백질)를 이용한 실험 결과, 알고리즘은 91개 단백질의 활성 사이트 주변의 바인딩 영역을 검출하였다. 이 논문은 단백질 활성 사이트를 분석하기 위한 새로운 기하학적 특징을 제시하였고, 활성 사이트와 관련이 없는 잔기를 제거함으로써 단백질 표면의 분석의 시간을 줄일 수 있는 장점이 있다.