• Biomedical Science Letters
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• v.4 no.1
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• pp.1-9
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• 1998

A Corynebacterium diphtheriae iron-repressible gene dirA, that was homologous to TSA of Saccharomyces cerevisiae and AhpC subunit of Salmonella typhimurium alkyl hydroperoxide reductase, was amplified with PCR and expressed in E. coli. The DirA purified from the transformed E. coli crude extracts prevented the inactivation of enzyme caused by metal-catalyzed oxidation (MCO) system containing thiols but not by ascorbate/Fe$^{3+}$/$O_2$ MCO system. The DirA concentration, which inhibited the inactivation of glutamine synthetase by 50% (IC$_{50}$) against MCO system, was 0.12 mg/ml. The multimeric forms of DirA were converted to the monomeric form in SDS-PAGE under the thioredoxin system comprised of NADPH, Saccharomyces cerevisiae thioredoxin reductase, and thioredoxin. Also, DirA showed thioredoxin dependent peroxidase activity. All of these results were consistent with the characteristics of a thiol specific antioxidant (TSA) protein having two conserved cysteine residues.

• The Journal of the Korean Society for Microbiology
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• v.8 no.1
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• pp.13-17
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• 1973

To understand the characteristics of 29 toxigenic strains of Corynebacterium diphtheriae isolated in Seoul area, type classification, biochemical properties and antibiotic susceptibility pattern to 9 kinds of antibiotics were investigated. The results obtained were summerized as follows; I. Among the 29 strains, gravis type was the overwhelming majority(24 strains), followed by intermedius type(3 strains) and mitis type(2 strains). II. Fermentation of glucose, maltose, lactose, trehalose and mannitol, nitrate reduction and urease were tested. All strains fermented glucose, but not sucrose, lactose, mannitol and trehalose. 9 strains fermented maltose and 20 strains did not. Nitrate was reduced by 28 strains but not by one strain. In urease test one strain showed positive, 28 strains negative. III. Antibiotic susceptibility test to penicillin G, chloramphenical, kanamycin, lincomycin, streptomycin, terramycin, erythromycin and gentamycin were carried out. The MIC of erythromycin(0.025 ${\mu}g/ml$ 26 strains and 0.05 ${\mu}g/ml$ 3 strains) was the lowest, followed by ampicillin, lincomycin and penicillin G. Streptomycin showed the highest MIC.

• KSBB Journal
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• v.14 no.2
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• pp.241-247
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• 1999

Experimental studies were carried out to optimize the culture conditions of Corynebacterium diphtheriae for the production of diphtheria toxin. A new media which does not contain any meat digest products was selected. The main ingredient of new medium was enzymatic digests of casein known as NZ-Case. In fermenter experiments, the toxin production was increased with the increase of cell growth. The optimum initial pH of media, air flow rate and agitation speed were 7.0, 0.22, vvm and 400 rpm, respectively. The contents of iron and calcium-phosphate precipitate were important for maximal cell growth and toxin production. The optimum concentration of iron was 0.3 mg/L and calcium-phosphate precipitate could serve in gradual supply of iron to maintain the optimal culture condition which is required for enhanced yield of toxin production. In potency test, the potency of toxoid from fermentor culture was higher than that from static culture. When diphtheria toxin is produced by fermentor culture, it is possible to produce higher levels of toxin and better toxoid quality in terms of safety, yield, productivity and immunity.

• YAKHAK HOEJI
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• v.22 no.2
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• pp.59-71
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• 1978

사람에게 병을 일으키는 수 많은 세균이 분비하는 다수의 외독소중, 이곳에서는 Corynebacterium diphtheriae의 외독소인 디프테리아 독소와 Vibrio cholerae가 분비하는 콜레라 장내 독소에 관해 주로 논하기로 하며, 디프테리아 독소와 생화학적 병인성 기전이 비슷한 Pseudomonas aeruginosa가 분비하는 pseudomnas외독소와 콜레라 장내독소와 병인성기전이 유사한 Escherichia coli의 장내독소에 관하여는 간략하게 언급하고져 한다.

• Molecules and Cells
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• v.38 no.8
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• pp.715-722
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• 2015

In Gram-negative bacteria in the periplasmic space, the dimeric thioredoxin-fold protein DsbC isomerizes and reduces incorrect disulfide bonds of unfolded proteins, while the monomeric thioredoxin-fold protein DsbA introduces disulfide bonds in folding proteins. In the Gram-negative bacteria Salmonella enterica serovar Typhimurium, the reduced form of CueP scavenges the production of hydroxyl radicals in the copper-mediated Fenton reaction, and DsbC is responsible for keeping CueP in the reduced, active form. Some DsbA proteins fulfill the functions of DsbCs, which are not present in Gram-positive bacteria. In this study, we identified a DsbA homologous protein (CdDsbA) in the Corynebacterium diphtheriae genome and determined its crystal structure in the reduced condition at $1.5{\AA}$ resolution. CdDsbA consists of a monomeric thioredoxin-like fold with an inserted helical domain and unique N-terminal extended region. We confirmed that CdDsbA has disulfide bond somerase/reductase activity, and we present evidence that the N-terminal extended region is not required for this activity and folding of the core DsbA-like domain. Furthermore, we found that CdDsbA could reduce CueP from C. diphtheriae.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.11-15
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• 2004

Embryonic stem (ES) cells are derived from the inner cell mass of the blastocyst-stage embryos that can be propagated indefinitely and, at the same time, can be differentiated into all the cell types that constitute the body. Current research using ES cells is mainly focused on the efficient generation of specific cell types by employing optimal differentiation conditions, which often requires the genetic manipulation of ES cells. As a way of developing an efficient system to regulate foreign gene expression in ES cells, we have inserted the gene encoding Corynebacterium diphtheria toxin-A (DTA) into an autonomously induced plasmid under positive doxycycline control ('Tet-on' system). In this study, we demonstrate that this system can lead to the cell death of mouse ES cells by the induction of DTA expression when exposed to the tetracycline derivative, doxycycline. MTT assay showed that this induction resulted in the apoptosis of ES cells.

• Biomaterials Research
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• v.22 no.4
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• pp.328-336
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• 2018

Background: Biogenic fabrication of silver nanoparticles from naturally occurring biomaterials provides an alternative, eco-friendly and cost-effective means of obtaining nanoparticles. It is a favourite pursuit of all scientists and has gained popularity because it prevents the environment from pollution. Our main objective to take up this project is to fabricate silver nanoparticles from lichen, Usnea longissima and explore their properties. In the present study, we report a benign method of biosynthesis of silver nanoparticles from aqueous-ethanolic extract of Usnea longissima and their characterization by ultraviolet-visible (UV-vis), Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) analyses. Silver nanoparticles thus obtained were tested for antimicrobial activity against gram positive bacteria and gram negative bacteria. Results: Formation of silver nanoparticles was confirmed by the appearance of an absorption band at 400 nm in the UV-vis spectrum of the colloidal solution containing both the nanoparticles and U. longissima extract. Poly(ethylene glycol) coated silver nanoparticles showed additional absorption peaks at 424 and 450 nm. FTIR spectrum showed the involvement of amines, usnic acids, phenols, aldehydes and ketones in the reduction of silver ions to silver nanoparticles. Morphological studies showed three types of nanoparticles with an abundance of spherical shaped silver nanoparticles of 9.40-11.23 nm. Their average hydrodynamic diameter is 437.1 nm. Results of in vitro antibacterial activity of silver nanoparticles against Staphylococcus aureus, Streptococcus mutans, Streptococcus pyrogenes, Streptococcus viridans, Corynebacterium xerosis, Corynebacterium diphtheriae (gram positive bacteria) and Escherichia coli, Klebsiella pneuomoniae and Pseudomonas aeruginosa (gram negative bacteria) showed that it was effective against tested bacterial strains. However, S. mutans, C. diphtheriae and P. aeruginosa were resistant to silver nanoparticles. Conclusion: Lichens are rarely exploited for the fabrication of silver nanoparticles. In the present work the lichen acts as reducing as well as capping agent. They can therefore, be used to synthesize metal nanoparticles and their size may be controlled by monitoring the concentration of extract and metal ions. Since they are antibacterial they may be used for the treatment of bacterial infections in man and animal. They can also be used in purification of water, in soaps and medicine. Their sustained release may be achieved by coating them with a suitable polymer. Silver nanoparticles fabricated from edible U. longissima are free from toxic chemicals and therefore they can be safely used in medicine and medical devices. These silver nanoparticles were stable for weeks therefore they can be stored for longer duration of time without decomposition.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.622-627
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• 2002

Diphtheria toxin repressor (DtxR) binds to approximately 30 to 35-bp regions containing an interrupted 9-bp inverted repeat within a 19-bp core sequence. The core sequence is fairly conserved and critical for DtxR binding. The flanking regions that are consisted of 5 to 8 more of nucleotides from the core are also required for DtxR binding. The nucleotides in both flanking regions are A-T rich. To examine whether the A-T nucleotides in both flanking regions from the core have significant roles for DtxR binding, a DNA fragment was constructed based on the diphtheria tox promoter/operator, and DNA fragments with substitution of A and T nucleotides In the flanking regions to G and C were also constructed. To assess the effect of these substitutions on binding of DtxR and repressibility by DtxR, $\beta$-galactosidase activity from lacZ fused to the region was assessed. Gel mobility shift of the region by purified DtxR was also examined. The DNA fragments containing the mutations in the flanking regions still exhibited repression and mobility shift with DtxR. The core segment with the mutation is still, therefore, recognized by DtxR. Nonetheless, the results from the assays indicated that the substitution significantly decreased repression of the operator by DtxR in vivo under high-iron condition and decreased binding of DtxR to the operator. These results suggest that A and T nucleotides fur both flanking regions are preferred for the binding of DtxR.

• KSBB Journal
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• v.26 no.6
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• pp.491-504
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• 2011

This review article provides an overview of the evolution of diphtheria vaccine, its value and its future. Diphtheria is an infectious illness caused by diphtheria toxin produced by pathogenic strains of Corynebacterium diphtheriae. It is characterized by a sore throat with membrane formation due to local tissue necrosis, which can lead to fatal airway obstruction; neural and cardiac damage are other common complications. Diphtheria vaccine was first brought to market in the 1920s, following the discovery that diphtheria toxin can be detoxified using formalin. However, conventional formalin-inactivated toxoid vaccines have some fundamental limitations. Innovative technologies and approaches with the potential to overcome these limitations are discussed in this paper. These include genetic inactivation of diphtheria toxoid, innovative vaccine delivery systems, new adjuvants (both TLR-independent and TLR-dependent adjuvants), and heat- and freeze-stable agents, as well as novel platforms for producing improved conventional vaccine, DNA vaccine, transcutaneous (microneedle-mediated) vaccine, oral vaccine and edible vaccine expressed in transgenic plants. These innovations target improvements in vaccine quality (efficacy, safety, stability and consistency), ease of use and/or thermal stability. Their successful development and use should help to increase global diphtheria vaccine coverage.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.7
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• pp.1017-1025
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• 2016

This study was performed in order to investigate changes in platycosides, as well as antimicrobial activities of bronchus diseases-inducing bacteria (Corynebacterium diphtheriae, Klebsiella pneumoniae, Staphylococcus aureus, and Streptococcus pyogenes) of Platycodon grandiflorum root (PGR) fermented by lactic acid bacteria (Leuconostoc mesenteroides N12-4, Leuc. mesenteroides N58-5, Lactobacillus plantarum N76-10, L. plantarum N56-12, Lactobacillus brevis N70-9, and L. brevis E3-8). Growth of L. plantarum on PGR was most active during lactic acid fermentation using different strains. Total platycoside, platycoside E, platycodin A, polygalacin $D_2$, polygalacin D, and diapioplatyco-side E contents of PGR fermented for 96 h at $37^{\circ}C$ by Leuc. mesenteroides and L. plantarum increased, whereas contents of platycodin D and platycodin $D_3$ were reduced. The antimicrobial activity on PGR fermented by L. plantarum N56-12 exhibited strong microbial proliferation for all four kinds of bronchus disease-inducing bacteria and was higher than that of non-fermented PGR extract. MIC of fermented PGR extract by L. plantarum N56-12 on C. diphtheriae, K. pneumoniae, S. aureus, and S. pyogenes were 45, 10, 50, and 25 mg/mL, respectively. Thus, this result shows that the antimicrobial activities of bronchus disease-inducing bacteria and platycoside content of PGR by L. plantarum N56-12 were higher than that of non-fermented PGR extract.