• Title/Summary/Keyword: MoFe 단백질

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Structural and Functional Analysis of Nitrogenase Fe Protein with MgADP bound and Amino Acid Substitutions (MgADP 결합 및 아미노산 치환 Nitrogenase Fe 단백질의 구조 및 기능 분석)

  • Jeong, Mi-Suk;Jang, Se-Bok
    • Journal of Life Science
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    • v.14 no.5
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    • pp.752-760
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    • 2004
  • The function of the [4Fe-4S] cluster containing iron (Fe-) protein in nitrogenase catalysis is to serve as the nucleotide-dependent electron donor to the MoFe protein which contains the sites for substrate binding and reduction. The ability of the Fe protein to function in this manner is dependent on its ability to adopt the appropriate conformation for productive interaction with the MoFe protein and on its ability to change redox potentials to provide the driving force required for electron transfer. The MgADP-bound (or off) conformational state of the nitrogenase Fe protein structure described reveals mechanisms for long-range communication from the nucleotide-binding sites to control affinity of association with the MoFe protein component. Two pathways, termed switches I and II, appear to be integral to this nucleotide signal transduction mechanism. In addition, the structure of the MgADP bound Fe protein provides the basis for the changes in the biophysical properties of the [4Fe-4S] observed when Fe protein binds nucleotides. The structures of the nitrogenase Fe protein with defined amino acid substitutions in the nucleotide dependent signal transduction pathways of the Switch I and Switch II have been determined by X-ray diffraction methods. These two pathways have been also implicated by site directed mutagenesis studies, structural analysis and analogies to other proteins that utilize similar nucleotide dependent signal transduction pathways. We have examined the validity of the assignment of these pathways in linking the signals generated by MgATP binding and hydrolysis to macromolecular complex formation and intermolecular electron transfer. The results provide a structural basis for the observed biophysical and biochemical properties of the Fe protein variants and interactions within the nitrogenase Fe protein-MoFe protein complex.

Mechanism of Biological Nitrogen Fixation in Azotobacter vinelandii (Azotobacter vinelandii에서의 생물학적 질소고정 작용 메카니즘)

  • Kim, Yong-Ung;Han, Jae-Hong
    • Applied Biological Chemistry
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    • v.48 no.3
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    • pp.189-200
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    • 2005
  • Biological nitrogen fixation is an important process for academic and industrial aspects. This review will briefly compare industrial and biological nitrogen fixation and cover the characteristics of biological nitrogen fixation studied in Azotobacter vinelandii. Various organisms can carry out biological nitrogen fixation and recently the researches on the reaction mechanism were concentrated on the free-living microorganism, A. vinelandii. Nitrogen fixation, which transforms atmospheric $N_2$ into ammonia, is chemically a reduction reaction requiring electron donation. Nitrogenase, the biological nitrgen fixer, accepts electrons from biological electron donors, and transfers them to the active site, FeMo-cofactor, through $Fe_4S_4$ cluster in Fe protein and P-cluster in MoFe protein. The electron transport and the proton transport are very important processes in the nitrogenase catalysis to understand its reaction mechanism, and the interactions between FeMo-cofactor and nitrogen molecule are at the center of biological nitrogen fixation mechanism. Spectroscopic studies including protein X-ray crystallography, EPR and $M{\ddot{o}}ssbauer$, biochemical approaches including substrate and inhibitor interactions as well as site-directed mutation study, and chemical approach to synthesize the FeMo-cofactor model compounds were used for biological nitrogen fixation study. Recent research results from these area were presented, and finally, a new nitrogenase reaction mechanism will be proposed based on the various research results.

Properties and Activities of Nireogenase System of Azospirillum amazonensa Kp1 (Azospirillum amazonense Kp1의 질소고정효소계의 활성 및 특성)

  • 송승달;김성준;추연식
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.151-157
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    • 1990
  • The maximum nitrogen fixation activity of the associative, microaerobic and acid tolerant bacteria, Azospirillum amazonense Kp1 was obtained with 0.2Kpa of $O_{2}$ and showed a reversible inhibition by the higher concentrations. Ammonium treatment caused a gradual inhibition of the activity up to 350mM. The nitrogenase systems were purified by gradient chromatography on DEAE-52 cellulose, heat treatment and preparative PAGE. The MoFe protein showed molecular weight of 210,000 including two nonidentical subunits with apparent molecular weights of 55,000 and 50,000 and an isoelectricpoint of 5.2 and contained 2, 24 and 28 atoms of Mo, Fe and acid labile S per molecule. The Fe protein revealed molecular weight of 66,000 including two types of subunits with molecular weights of 35,000 and 31,000 and an isoelectric point of 4.6, and contained 4 atoms of Fe and 6 atoms of S per molecule. The maximum specific nitrogenase activity attained 2,200 and 1,700nM $C_2H_4mg^{-1} min^{-1}$, respectively for MoFe and Fe proteins at pH7 and $35^{\circ}C$. The activity was lost after 10 and 30 days under the cold room ($4^{\circ}C$) condition for Fe and MoFe proteins, respectively.

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Effects of Various Diets on Growth and Body Composition of Juvenile Parrot Fish, Oplegnathus fasciatus (먹이 종류가 돌돔 Oplegnathus fasciatus 치어의 성장 및 체성분에 미치는 영향)

  • Moon Lee, HaeYoung;Nam, Myung-Mo
    • Korean Journal of Ichthyology
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    • v.27 no.4
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    • pp.293-299
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    • 2015
  • The feeding experiment was conducted to investigate the effects of one experimental diet (EDP) and five different commercial diets (CEPs) on growth and body composition for juvenile parrot fish, Oplegnathus fasciatus. An EDP was formulated to contain 50% crude protein (CP) from fishmeal, casein, zein and wheat flour and 15% crude lipid (CL) from squid liver oil. Five CEPs for seawater fish were two domestic E commercial diet (DECD) and C commercial diet (DCCD), three imported H commercial diet (IHCD), L commercial diet (ILCD) and O commercial diet (IOCD) containing 53.1~66.6% CP and 10.7~14.6% CL, respectively. Each diet was fed to triplicate groups of juvenile parrot fish initially weighing $1.14{\pm}.01g/fish$ (mean${\pm}$SD) in a flow-through seawater system with a water temperature of $19.0{\sim}25.0^{\circ}C$. Weight gain (WG) and feed efficiency (FE) were significantly greatest in fish fed the DCCD and IOCD; intermediate responses were observed for fish fed the ILCD, while the IECD, IHCD, and the EDP produced the lowest WG and FE values. Survival with no significant difference approached 100% for fish fed the all six diets in this experiment. Whole-body moisture, protein, lipid and ash contents were not affected by the different type of diets. Therefore, type of diets appeared to be important factor in influencing WG and FE of juvenile parrot fish; the best diets for juvenile parrot fish was determined to be the domestic commercial C and the imported commercial O diets containing high protein (61.3, 66.6%) and lipid (14.6, 13.0%) in natural seawater based on highest WG, and FE, respectively. This study indicates that the two commercially formulated diets containing two highest proteins and lipids used in this experiment could be practical diets for juvenile parrot fish; these differences of growth performance between experimental diet and commercial diets may be reason for different dietary protein and lipid levels.

The Medium Optimization through Continuous Culture of an Methanol Utilizing Bacterium for SCP Production (SCP 생산용 메탄올 자화균주의 연속배양에 의한 배지 최적화)

  • 김창호;김태진홍석인
    • KSBB Journal
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    • v.5 no.4
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    • pp.355-363
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    • 1990
  • Methanol-utilizing bacterium isolated from sewage samples in Seoul showed optimal temperature and pH of $33^{\circ}C$ and 7.1 for growth, respectively. The maximum specific growth rate was $0.42hr^{-1}$. The minimum medium composition was reconstituted depending on the surplus and the deficit of each component in the basal medium at steady state. The optimal composition was given as(g/l); Methanol 40, $(NH_4)_2\;SO_42, \;KH_2PO_4\;1.5, \;K_2HPO_4\;0.2, \;H_3PO_4\;0.79, \;Na_2HPO_4{\cdot}12H_2O\;0.15, \;MgSO_4{\cdot}7H_2O\;1.5, \;FeSO_4{\cdot}7H_2O\;0.034, \;MnSO_4{\cdot}4H_2O\;0.005, \;CuSO_4{\cdot}5H_2O\;0.0027, \;CaCl_2{\cdot}2H_2O\;0.25, \;ZnSO_4{\cdot}7H_2O\;0.007, \;(NH_4)_6\;Mo_7O_{24}{\cdot}4H_2O\;0.00048, \;H_3BO_3\;0.00068, \;CoCl_2\; 0.00024$ Under the continuous culture with optimum medium the maximum cell productivity was 3.8g/1/hr at dilution rate $0.23hr^{-1}$. Maximum cell concentration and its protein content were 19.5g/l and 70% at dilution rate of $0.1hr^{-1}$, respectively.

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Effects of Soil pH on Nutritional and Functional Components of Chinese Cabbage (Brassica rapa ssp. campestris) (토양 pH가 배추(Brassica rapa ssp. campestris)의 영양성분과 기능성분에 미치는 영향)

  • Lee, Jo-Eun;Wang, Pingjuan;Kim, Gyung-Yun;Kim, Sung-Han;Park, Su-Hyoung;Hwang, Yong-Soo;Lim, Yong-Pyo;Lee, Eun-Mo;Ham, In-Ki;Jo, Man-Hyun;An, Gil-Hwan
    • Horticultural Science & Technology
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    • v.28 no.3
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    • pp.353-362
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    • 2010
  • The contents of functional and nutritional components of 13 cultivars of Chinese cabbage (CC, $Brassica$ $rapa$ subspecies $campestris$) were analyzed to compare the effects of soil pH of the greenhouse (pH 6.2) and outdoor (pH 7.6). The CC cultivated on pH 6.2 (CC-6.2) soil contained significantly increased amounts (2-9 fold) of pectin, crude protein, vitamin C and vitamin E compared to the counterpart (CC-7.6). The contents of ash and the minerals (Ca, Fe, Na, and Mn) were also significantly increased in CC-6.2. However, CC-6.2 contained 40-50% lower contents of reducing sugars, cellulose and crude fat than CC-7.6. CC-7.6 contained more glucosinolates, gluconasturtiin (18.33 vs. $1.16nmol{\cdot}g^{-1}$ wet weight) and gluconapin (145 vs. $2nmol{\cdot}g^{-1}$ wet wt), than CC-6.2. In conclusion, CC-6.2 had an improved texture (high pectin and low cellulose) and nutritional value (high in protein, Ca, Fe, vitamin C, and E), whereas the CC-7.6 had better taste (high in reducing sugars) and anticancer functionality (high in glucosinolates).

Effect of Rearing Period on Chemical Composition of Duck Meats (출하 일령이 오리육의 화학적 변화에 미치는 영향)

  • Chae Hyun-Seok;Yoo Young-Mo;Ahn Chong-Nam;Kim Dong-Hun;Ham Jun-Sang;Jeong Seok-Keun;Lee Jong-Moon;Choi Yang-Il
    • Food Science of Animal Resources
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    • v.26 no.1
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    • pp.9-14
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    • 2006
  • The aim of this study was to determine the chemical traits of duck meats with different rearing periods(45 70 days). The content of protein was 21.3 and 19.61% in the breast and leg at 70 days post hatched, respectively. Group at 70 days showed high protein composition than those of 45 days group. The content of fat was slightly increased with the rearing periods. The composition of Ca, mineral compound, was not showed the difference between leg and breast meat. However, it was increased with the rearing periods. The composition of Fe was not significantly different in the breast meat. However, there was a significant increase in the leg (p<0.05). It was compared 14.6 ppm at 45 days group to 27.5 ppm at 70 days group. The content of methionine was significantly increased in the breasts due to the increasing rearing periods (p<0.05), but not in leg meat. There was no significant difference between parts, but glutamic acid content was significantly decreased along with the increasing rearing periods (p<0.05). The content of collagen at 70 days(0.77 g/100 g) was higher than those of 45 days(0.65 g/100 g). It was affected by the rearing periods In the wing-removed skit, it was 0.15 g increases at 70 rearing period group. The cholesterol content was significantly increased in the leg but not in the breast. With the rearing periods, it was significantly increased in the leg (p<0.05).