• Korean Journal of Soil Science and Fertilizer
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• v.49 no.1
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• pp.17-29
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• 2016

Though there is an abundant supply of nitrogen in the atmosphere, it cannot be used directly by the biological systems since it has to be combined with the element hydrogen before their incorporation. This process of nitrogen fixation ($N_2$-fixation) may be accomplished either chemically or biologically. Between the two elements, biological nitrogen fixation (BNF) is a microbiological process that converts atmospheric di-nitrogen ($N_2$) into plant-usable form. In this review, the genetics and mechanism of nitrogen fixation including genes responsible for it, their types and role in BNF are discussed in detail. Nitrogen fixation in the different agricultural systems using different methods is discussed to understand the actual rather than the potential $N_2$-fixation procedure. The mechanism by which the diazotrophic bacteria improve plant growth apart from nitrogen fixation such as inhibition of plant ethylene synthesis, improvement of nutrient uptake, stress tolerance enhancement, solubilization of inorganic phosphate and mineralization of organic phosphate is also discussed. Role of diazotrophic bacteria in the enhancement of nitrogen fixation is also dealt with suitable examples. This mini review attempts to address the importance of diazotrophic bacteria in nitrogen fixation and plant growth improvement.

• 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.

• Korean Journal of Soil Science and Fertilizer
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• v.20 no.3
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• pp.261-268
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• 1987

A green house experiment was conducted to find out the differences in the amount of biologically fixed nitrogen and kjeldahl nitrogen on the different soil texture, kinds and amounts of fertilizer nitrogen under light (photosynthetic $N_2$-fixation) and dark (heterotrophic $N_2$-fixation) condition in submerged paddy soil. The reults obtained were summarized as follows; 1. The amount of biologically fixed nitrogen per mg carbon from different organic matter was obtained as 0.13 mg in glucose, 0.09 mg in rice straw, and 0.07 mg in refused mushroom compost and barley straw under 60 days of incubation. 2. Nitrogen fixing activities were decreased with increase of fertilizer nitrogen and those tendency was pronounced more in sandy soil with application of urea than that of ammonium sulfate. 3. The application of ammonium sulfate in sandy soil under light condition was increased the photosynthetic $N_2$-fixation and the applied urea was remarkably reduced the heterotrophic $N_2$-fixation in sandy soil. The proportion of biologically fixed total nitrogen after experiment in sandy soil was obtained as 25% for dark(heterotrophic $N_2$-fixation) and 75% for light (photosynthetic $N_2$-fixation) condition. On the other hand, very similar biological $N_2$-fixing tendency was obtained between kinds of nitrogen fertilizer and two light condition in clayey soil. 4. The kjeldahl nitrogen was remarkably decreased after experiment under dark condition with application of urea than that of light condition with ammonium sulfate, and no remarkable decreasing tendency was obtained in clayey soil between two kinds of fertilizer nitrogen. 5. The high significant positive correlationship was obtained between calculated biological nitrogen fixation by acetylene reducing activity and kjeldahl nitrogen after experiment under light (y=0.8488X-5.9632, $r=0.9928^{**}$, n=21) and dark (y=0.8795X-7.1056, $r=0.9782^{**}$, n=21) condition. In this experiment condition, conversion factors of 6:1 was obtained from biological nitrogen fixation to soil nitrogen.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.1
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• pp.1-4
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• 2005

This experiment was conducted to investigate the soil nitrogen credit of biological nitrogen fixation (BNF) and the nitrogen balance of soybean in soybean-barley cropping systems. Soybean cultivar, Shinpaldalkong2 and barley cultivar, Olbori, were used in soybean mono-cropping (SM), barley monocropping (BM), and barley­soybean double cropping system. The barley-soybean double cropping system was treated with two different levels of nitrogen fertilizers, 0 nitrogen fertilizer (BS-F0), and standard nitrogen fertilizer (BS-F1). Nitrogen and organic matter concentrations in soil of BS-F1 plot on October, 2001 were increased $4.8\%\;and\;5.9\%$, respectively, compared with those on October, 2000. The ranges of BNF rate in soybean were $69.1\~ 88.2\%$ in two years, and the rate was the highest in BS-F0 plot and the lowest in SM plot. The ranges of nitrogen harvest index (NHI) in all treatments were $83.9\~86.7\%$. The yield was 270 kg/10a in BS­F1 plot and 215 kg/10a in BS-F0 plot. However, the nitrogen balances were +0.6 kg/10a of gain of soil nitrogen in BS-F0 plot and -0.4 kg/10a of loss of soil nitrogen in BS-F1 plot. In comparisons of SM and BS-F1 plots, although the seed yields were similar in two plots, the loss of soil nitrogen was higher in SM than BS-F1 plot. Overall, our results suggest that barley-soybean double cropping system was more effective in respect to seed productivity and soil nitrogen conservation than soybean monocropping system, and the N credit to following crops by soybean cultivation was identified in soybean double cropping system.

• Environmental Engineering Research
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• v.12 no.4
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• pp.136-147
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• 2007

In an effort to identify possible microbes for seeking bioagents for remediation of herbicide-contaminated soils, seven species of phototrophic nonsulfur bacteria (Rhodobacter capsulatus and sphaeroides, Rhodospirillum rubrum, Rhodopseudomonas acidophila, blastica and viridis, Rhodomicrobium vannielii) were grown in the presence of the herbicide, butachlor, and bacterial growth rates and nitrogen fixation were measured with different carbon sources. Under general conditions, all species showed 17-53% reductions in growth rate following butachlor treatment. Under nitrogen-fixing conditions, Rb. capsulatus and Rs. rubrum showed 1-4% increases in the growth rates and 2-10% increases in nitrogen-fixing abilities, while the other 5 species showed decreases of 17-47% and 17-85%, respectively. The finding that Rp. acidophila, Rp. blastica, Rp. viridis and Rm. vannielii showed stronger inhibitions of nitrogenase activity seems to indicate that species in genera Rhodobacter and Rhodospirillum are less influenced by butachlor than those in Rhodopseudomonas and Rhodomicrobium in terms of nitrogen-fixing ability. Overall, nitrogenase activity was closely correlated with both growth rate and glutamine synthetase activity (representing nitrogen metabolism). When the carbon sources were compared, pyruvate (three carbons) was best for all species in terms of growth rate and nitrogen fixation, with malate (four carbons) showing intermediate values and ribose(five carbons) showing the lowest; these trends did not change in response to butachlor treatment. We verified that each of the 7 species had a plasmid ($12.2{\sim}23.5\;Kb$). We found that all 7 species could use butachlor as a sole carbon source and 3 species were controlled by plasmid-born genes, but it is doubtful whether plasmid-born genes were responsible to nitrogen fixation.

• Korean Journal of Organic Agriculture
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• v.19 no.spc
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• pp.161-164
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• 2011

Nitrogen fixation by legumes can be valuable sources for organic farming. This study was to investigate the effect of different legume mixtures on nitrogen fixation and transfer to grasses on spring paddy field. Three different mixtures were used (rye+hairy vetch, Italian ryegrass+crimson clover, oat+pea) in a randomized complete block design with three replications and sowed in pots with different sowing rate (5:5 rye:hairy vetch,7:3=Italian:crimson, 6:4=oat:pea) on early March. $(^{15}NH_4)SO_4$ solution at. 99.8 atom%$^{15}N$ was applied to the each pot at the rate of 2kg N $ha^{-1}$ on $16^{th}$ April. Forage were harvested at ground level in heading stage and separated into legume and grass. Total N content and $^{15}N$ value were determined using a continuous flow stable isotope ratio mass spectrometry. DM yield of rye+vetch, Italian+crimson and oat+pea were 6,607, 3,213 and 4,312kg/ha, respectively. Proportion of N from fixation was 0.73(rye+vetch), 0.42(Italian+crimson) and 0.93(oat+pea). The percentages of N transfer from legume to grass were from 61% to 24% in different method by treatments and -35% to 21% in isotope dilution method.

• Proceedings of the Korean Society of Crop Science Conference
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• 2001.09a
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• pp.152-153
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• 2001

• Journal of Microbiology and Biotechnology
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• v.25 no.8
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• pp.1339-1348
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• 2015

Until now, considerable effort has been made to engineer novel nitrogen-fixing organisms through the transfer of nif genes from various diazotrophs to non-nitrogen fixers; however, regulatory coupling of the heterologous nif genes with the regulatory system of the new host is still not well understood. In this work, a 49 kb nitrogen fixation island from P. stutzeri A1501 was transferred into E. coli using a novel and efficient transformation strategy, and a series of recombinant nitrogen-fixing E. coli strains were obtained. We found that the nitrogenase activity of the recombinant E. coli strain EN-01, similar to the parent strain P. stutzeri A1501, was dependent on external ammonia concentration, oxygen tension, and temperature. We further found that there existed a regulatory coupling between the E. coli general nitrogen regulatory system and the heterologous P. stutzeri nif island in the recombinant E. coli strain. We also provided evidence that the E. coli general nitrogen regulator GlnG protein was involved in the activation of the nif-specific regulator NifA via a direct interaction with the NifA promoter. To the best of our knowledge, this work plays a groundbreaking role in increasing understanding of the regulatory coupling of the heterologous nitrogen fixation system with the regulatory system of the recipient host. Furthermore, it will shed light on the structure and functional integrity of the nif island and will be useful for the construction of novel and more robust nitrogen-fixing organisms through biosynthetic engineering.

• Applied Chemistry for Engineering
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• v.21 no.3
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• pp.243-251
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• 2010

Nitrogen oxides (NOx) in combustion flue gas are currently mitigated by chemical processes such as catalytic reduction, absorption and adsorption. However, development of environmentally sustainable biological processes is necessary in the near future. In this paper, the up-to-dated R&D trend of biological methodologies regarding NOx removal was reviewed, and their advantages and disadvantages were discussed. The principles and applications of bacterial system including nitrification and denitrification and photosynthetic microalgae system were compared. In order to enhance biological treatment rate and performance, the insoluble nitric oxide (NO) should be first absorbed using a proper solubilization agent, and then microbial degradation or fixation is to be followed. The use of microalgal system has a good prospect because it can fix $CO_2$ and NOx simultaneously and requires no additional carbon for energy source.

• Applied Biological Chemistry
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• v.24 no.2
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• pp.132-138
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• 1981

Four species of the photosynthetic bacteria from 36 samples were isolated. their characteristics, their capability for nitrogen fixation and their capability for purification of organic waste water by photosynthetic bacteria were examined. Photosynthetic bacteria are widely distributed in soil. The isolated species are Rhodopseudomonas capsulatus. R. spheroides, R. gelatinosa, and R. ruburm. The capability for nitrogen fixation varies with the species of photosynthetic bacteria, and it is very pronounced in the R. capsulatus. The capability for purification of organic waste water is relatively strong but varies with the species.