• Journal of the Korean Magnetic Resonance Society
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• v.19 no.3
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• pp.106-111
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• 2015

Helicobacter pylori (H. pylori) survives in acidic and fluctuating pH conditions of the stomach. The pH effect on H. pylori proteins is important for the advanced understanding of its evolution and viability, although this bacterium has the molecular machinery that neutralizes the acidic condition. HP1492 is known as a conserved NifU-like protein from H. pylori. NifU is a nitrogen fixation protein that mediates the transfer of iron-sulfur (Fe-S) cluster to iron-sulfur proteins like ferredoxin. Commonly, the monomeric reduced state of NifU can be converted to the dimeric oxidized state by intermolecular disulfide bond formation. Because it remains unclear that HP1492 actually behaves as known NifU protein, we first found that this protein can adopt both oxidized and reduced forms using size exclusion chromatography. Circular dichroism experiment showed that HP1492 is relatively well-structured at pH 6.5, compared to other pH conditions. On the basis of the backbone resonance assignment of HP1492, we further characterized the residues that are sensitive to pH using NMR spectroscopy. These residues showing large chemical shift changes could be mapped onto the secondary structure of the protein. Our results could provide the foundation for structural and biophysical studies on a wide spectrum of NifU proteins.

• The Korean Journal of Food And Nutrition
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• v.5 no.2
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• pp.137-143
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• 1992

As a research for developing biofertilizers, Klebsiella oxytoca, an associative nitrogen fixer in the rhizosphere of rice plant in the soil of paddy field, was subjected to molecular breeding. The results obtained were as followings. 1). By transforming pbIC71A, Nif A overproducing plasmid, into Klebsiella oxytoca NGl3, Klebsiell6f oxytoca SH3l, and Klebsiella oxytoca SH161, nitrogenase activities in the absence of nitrogen source in the medium were increased 6.4, 17.2, and 13.5 times, respectively, in comparison with the parent strains. 2). Nitrogenase activity of Klebgiella oxytoca NGl3, Klebsiella oxytoca SH3l, and Klebsiella oxytoca SH161 was completely repressed In the presence of 15mM NH4+. But, nitrogenase activities of Klebsiella oxytoca NGl3/PMC71A, Klebsiella oxytoca SH3l /PMC71A, and Klebsiella oxytoca SH 161/pMC714 harboring PMC71A, were 13.7%, 7.7%, and 6.2% of the nitrogenase activities in the absence of nitrogen source in the medium, respectively.

• Korean Journal of Microbiology
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• v.15 no.3
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• pp.131-138
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• 1977

The effects of various nitrogen soruces on the expression of nif gene were investigated using nif-lac fusants of Klebsiella pneumoniae. K. pneumoniae UK 2979 was infected with Mudl lysate prepared by heat induction of K. pneumoniae UK 4482. About 80 nif-lac fusants were greatly repressed. Amino acids, such as serine, glutamine and asparagine, were found to support the growth of K. pneumoniae M5al quite well, and showed a repressive effect on .betha.-galactosidase activities of nif-lac fusants LX-9 and LX-22 in NFHM. Glutamic acid, histidine and arginine rendered poor growth but high activities of .betha.-galactosidase. Good cell growth and high enzyme activity were observed when complex nitrogen sources, such as casitone, proteose pepone, were employed. .betha.-Galactosidase activities of LX-9 and LX-22 in nitrogen free minimal medium increased sharply within first 4 hours.

• The Korean Journal of Physiology and Pharmacology
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• v.11 no.1
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• pp.21-30
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• 2007

The present study was designed to establish comparatively the inhibitory effects of cilnidipine(CNP), nifedipine(NIF), and $\omega$-conotoxin GVIA(CTX) on the release of CA evoked by cholinergic stimulation and membrane depolarization from the isolated perfused model of the rat adrenal medulla. CNP(3 ${\mu}M$), NIF(3 ${\mu}M$), and CTX(3 ${\mu}M$) perfused into an adrenal vein for 60 min produced greatly inhibition in CA secretory responses evoked by ACh($5.32{\times}10^{-3}M$), DMPP($10^{-4}M$ for 2 min), McN-A-343($10^{-4}M$ for 2 min), high $K^+(5.6{\times}10^{-2}M)$, Bay-K-8644($10^{-5}M$), and cyclopiazonic acid($10^{-5}M$), respectively. For the CA release evoked by ACh and Bay-K-8644, the following rank order of potency was obtained: CNP>NIF>CTX. The rank order for the CA release evoked by McN-A-343 and cyclopiazonic acid was CNP>NIF>CTX. Also, the rank orders for high $K^+$ and for DMPP were NIF>CTX>CNP and NIF>CNP>CTX, respectively. Taken together, these results demonstrate that all voltage-dependent $Ca^{2+}$ channels(VDCCs) blockers of cilnidipine, nifedipine, and $\omega$-conotoxin GVIA inhibit greatly the CA release evoked by stimulation of cholinergic(both nicotinic and muscarinic) receptors and the membrane depolarization without affecting the basal release from the isolated perfused rat adrenal gland. It seems likely that the inhibitory effects of cilnidipine, nifedipine, and $\omega$-conotoxin GVIA are mediated by the blockade of both L- and N-type, L-type only, and N-type only VDCCs located on the rat adrenomedullary chromaffin cells, respectively, which are relevant to $Ca^{2+}$ mobilization. It is also suggested that N-type VDCCs play an important role in the rat adrenomedullary CA secretion, in addition to L-type VDCCs.

• 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.

• Korean Journal of Microbiology
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• v.26 no.3
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• pp.181-187
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• 1988

Using a Tn5-Mob system, pEA9 was characterized as a self-transmissible plasmid carrying a kanamycin resistance marker. The self-transfer frequencies of pEA9 varied greatly depending on pH values. The transfer frequency was about $4\times 10^{-5}$ at pH 5, that was 10 times higher than one at pH 6.5. With a helper plasmid, transfer frequencies were increased about $10^{4}$ times than the frequencies obtained without it.

• Journal of the Korean Magnetic Resonance Society
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• v.17 no.2
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• pp.105-110
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• 2013

HP1492 is a NifU-like protein of Helicobacter pylori (H. pylori) and plays a role as a scaffold which transfer Fe-S cluster to Fe-S proteins like Ferredoxin. To understand how to bind to iron ion or iron-sulfur cluster, HP1492 was expressed and purified in Escherichia coli (E. coli). From the NMR measurement, we could carry out the sequence specific backbone resonance assignment of HP1492. Approximately 91% of all resonances could be assigned unambiguously. By analyzing results of CSI and TALOS from NMR data, we could predict the secondary structure of HP1492, which consists of three ${\alpha}$-helices and three ${\beta}$-sheets. This study is an essential step towards the structural characterization of HP1492.

• Korean Journal of Microbiology
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• v.22 no.4
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• pp.265-269
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• 1984

For the utilization as donor cells of conjugation, DAP-Auxotrophs were isolated from C. coli cells, carrying plasmid $p^{RD1}$ with(a) drug resistance makers from Pseudononas $(Km^r, \;Carb^r, \;Tc^r)$ and (b) the nif-gene group from Klebsiela. E. coli $p^{RD1}$ cells were treated with nitrosoguanidine for the mutagenesis and cephalexin for the isolation of DAP-Auxotrophs. The nature of auxotrophs was verified by suitable biochemical test and checking with 6-cyanopurine as a color indicator for the presence of nif-gene.

• Proceedings of the Zoological Society Korea Conference
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• 1997.10b
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• pp.271.1-271
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• 1997

No Abstract, See Full Text

• Journal of Korean Clinical Health Science
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• v.7 no.1
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• pp.1232-1237
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• 2019

Purpose. In this study, two types of soft contact lenses with different materials were selected to compare the time of tear film destruction and high order aberrations before and after wearing. Methods. Thirty patients (60 eyes) in their 20s were included in this study. Two lenses with different materials, Group 4 (Etafilcon A) and Group 5 (Narafilcon A) were selected. Using aberration analyzer and keratometry, high-order aberration and tear film test (NIF-BUT, NIAvg-BUT) were performed before and after wearing. Results. When comparing the higher aberrations of the Etafilcon A and Narafilcon A lenses, the higher aberrations of the Narafilcon A lens were higher overall. For the initial tear film break-up time (NIF-BUT) after wearing, the Etafilcon A lens was reduced by 4.0 seconds and the Narafilcon A lens increased by 0.6 seconds. For the mean tear film break-up time (NIAvg-BUT) after wearing, the Etafilcon A lens decreased by 2.4 seconds and the Narafilcon A lens increased by 1.7 seconds. Conclusions. NIF-BUT and NIAvg-BUT of Narafilcon A lens were increased. The lens with relatively low water content and higher oxygen permeability than the lens with high water content has relatively less tear evaporation, which means that the time of destruction of the tear film is increased.