• Journal of Life Science
• /
• v.14 no.5
• /
• pp.752-760
• /
• 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.

• Applied Biological Chemistry
• /
• v.48 no.3
• /
• pp.189-200
• /
• 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.

• Proceedings of the Korean Biophysical Society Conference
• /
• 2002.06b
• /
• pp.18-18
• /
• 2002

Nitrogenase, comprised of the MoFe and Fe proteins, catalyzes the reduction of dinitrogen to ammonia at ambient temperature and pressure. The MoFe protein contains two metal centers, the P-cluster (Fe8S7-8) and the FeMo-cofactor (Fe7S9：homocitrate), the substrate binding site. Despite the availability of the crystal structure of the MoFe protein, suprisingly little is known about the molecular details of catalysis at the active site, and no small-molecule substrate or inhibitor had ever been shown to directly interact with a protein-bound cluster of the functioning enzyme, until our electron-nuclear double resonance(ENDOR) study of CO-inhibited nitrogenase.(omitted)

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.24 no.1
• /
• pp.1-10
• /
• 2004

This pot experiment was conducted to investigate the effects of combined micronutrient application($T_1$;control, $T_2$; Fe, $T_3$; Fe＋Mn, $T_4$; Fe＋Mn＋Cu, $T_5$ ; Fe＋Mn＋Cu＋Zn, $T_6$ ; Fe＋Mn＋Cu＋Zn＋Mo, $T_7$ ; Fe＋Mn＋Cu＋Zn＋Mo＋B) on forage performance of pure and mixed cultures of orchardgrass and white clover. The third part was concerned with the changes in the contents and yields of N-compounds (crude/pure protein and soluble N-compounds) in forages. The results obtained are summarized as follows: 1. The contents of N-compounds(crude/pure protein and soluble N-compounds) were generally different according to the forage species, whether it was a pure or mixed culture, and additional fertilization, especially N. In orchardgrass, these contents were relatively low at the $T_3$ and $T_6$ in both pure and mixed cultures. In white clover, these contents were relatively decreased by the $T_1$, $T_3$, and $T_6$ in mixed culture. 2. The treatments influenced relatively more on the yields of crude/pure protein than on the dry matter yields of forages, and this tendency was more significant in white clover than in orchardgrass. 3. In white clover, the great differences in the yields of crude protein by the treatments occurred especially in mixed culture and at 5th cut without no additional fertilization. In white clover, the positive effects of optimum treatments on the yields of crude protein seemed to be decreased by the additional fertilization, especially N. In mixed culture, the favorable growth of white clover by the optimum treatments tended to be positively related to the favorable contents and yields of N-compounds. The changes in the yields of pure protein were similar to the tendency of crude protein

• Korean Journal of Microbiology
• /
• v.28 no.2
• /
• pp.151-157
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.2
• /
• pp.294-300
• /
• 2010

The Iro protein is a member of the HiPIP family with the [$Fe_4S_4$] cluster for electron transfer. Many reports proposed that the conserved aromatic residues might be responsible for the stability of the iron-sulfur cluster in HiPIP. In this study, Tyr10 was found to be a critical residue for the stability of the [$Fe_4S_4$] cluster, according to site-directed mutagenesis results. Tyr10, Phe26, and Phe48 were essential for the stability of the [$Fe_4S_4$] cluster under acidic condition. Trp44 was not involved in the stability of the [$Fe_4S_4$] cluster. Molecular structure modeling for the mutant Tyr10 proteins revealed that the aromatic group of Tyr10 may form a hydrophobic barrier to protect the [$Fe_4S_4$] cluster from solvent.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.2
• /
• pp.124-128
• /
• 2011

Ferredoxin is a typical iron-sulfur protein that is ubiquitous in biological redox systems. This study investigates the in vitro assembly of a [$Fe_2S_2$] cluster in the ferredoxin from Acidithiobacillus ferrooxidans in the presence of three scaffold proteins: IscA, IscS, and IscU. The spectra and MALDI-TOF MS results for the reconstituted ferredoxin confirm that the iron-sulfur cluster was correctly assembled in the protein. The inactivation of cysteine desulfurase by L-allylglycine completely blocked any [$Fe_2S_2$] cluster assembly in the ferredoxin in E. coli, confirming that cysteine desulfurase is an essential component for iron-sulfur cluster assembly. The present results also provide strong evidence that [$Fe_2S_2$] cluster assembly in ferredoxin follows the AUS pathway.

• Journal of Nutrition and Health
• /
• v.17 no.1
• /
• pp.31-40
• /
• 1984

A new microassay was proposed for the determination of serum thiobarbituric acid(TBA) value greatly enhanced by ferric ion under anaerobic condition. One ${\mu}mole$ of $FeCl_{3}$ per $10{\mu}l$ of serum was added to the TBA reaction mixture containing serum protein precipitate. The reaction mixture was heated on boiling water-bath for 50min under $N_{2}$ flushing. The sensitivity of this assay was greatly enhanced by 40 times comparing with that of Yagi's method (1976). In favour of the enhancement, this test could be measured by colorimetry or spectrophotometry with the sample size of $10-20{\mu}l$ serum. The sensitivity and reproducibility were also improved by means of partial dehydration of the butanol extract with $(NH_{4})_{2}SO_{4}$ salting-out. Serum TBA values of healthy human at different age groups were determined by this proposed method.

• Korean Journal of Agricultural Science
• /
• v.37 no.1
• /
• pp.73-80
• /
• 2010

Functional and nutritional components of 7 cultivars of Chinese cabbage (CC; Brassica rapa subspecies campestris) from China were analyzed to compare the effects of soil pH (6.9 vs. 7.6). The CC grew on the soil of pH6.9 (CC-6.9) contained significantly increased amounts (2.3~4-fold) of pectin, crude protein, vitamin C and vitamin E compared to the control. The contents of ash and minerals (Ca, Fe, Na, Zn, K and Mn) were also significantly increased in CC-6.9. However, CC-6.9 contained 16-75% lower contents of reducing sugars, cellulose and crude fat than CC-7.6. CC-7.6 contained more glucosinolates than CC-6.9; gluconasturtiin (20.68 vs. 0.84 $nmol{\cdot}g^{-1}$ wet wt) and gluconapin (202.55 vs. 0.15 $nmol{\cdot}g^{-1}$ wet wt). In conclusion, CC-6.9 had an improved texture (high pectin and low cellulose) and nutritional value (high in protein, Ca, Fe, Zn, vitamin C, and E), whereas CC-7.6 had better taste (high in reducing sugars) and anticancer functionality (high in glucosinolates).

• Korean Journal of Ichthyology
• /
• v.27 no.4
• /
• pp.293-299
• /
• 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.