• Korean Journal of Microbiology
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• v.42 no.1
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• pp.40-46
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• 2006

Galactose joined to glucose by a $\beta(1\rightarrow4)$ glycosidic bond makes lactose and this disaccharide is rich in milk. It is known that lacotse is hydrolyzed to each monomeric sugar by either lactase in human or $\beta-galactosidase$ in bacteria. Ingestion of milk by lactase-deficient persons causes a temporary diarrhea and subsequent chronic diarrhea results in colitis with chronic inflammation. We isolated a $\beta-galactosidase$ producing psycrotolerant strain AS-20 from near cattle shed and investigated the growth at various temperature conditions. Whereas Escherichia coli strains did not grow at $10^{\circ}C$, the AS-20 strain could grow well at this low temperature and showed optimal growth at $30^{\circ}C$. The isolated strain was identified as 97% Hafnia alvei by biochemical properties. This strain could ferment glucose, lacotse, maltose, mannitol, xylose, ONPG, rhamanose and L-arabinose, and decarboxylate lysin and ornithine. To confirm the identity of isolated strain we amplified 16S rDNA by PCR and searched similarity of the 1426 bp DNA sequcence with Genbank database. The strain AS-20 showed 99% similarity with Hafnia alvei. The activity of $\beta-galactosidase$ was 1.5 times higher when the cell was grown at 10 or $20^{\circ}C$ than at $30^{\circ}C$. The highest enzyme activity of AS-20 was also much higher than that of E. coli, which was grown at $30^{\circ}C$.

• Journal of Oral Medicine and Pain
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• v.33 no.2
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• pp.159-166
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• 2008

An ideal anti-bacterial medication for oral infection requires selective effect on pathogens causing dental caries and periodontal disease but not on normal flora. In addition, it should be less toxic for human and even for environment. This study was to seek such a natural anti-bacterial medication and thus anti-bacterial effect of Hamamelis virginiana was evaluated. Many recent researches on the anti-bacterial effect of natural plant extract and essential oil have reported that natural products can be used as medication for prevention and restrainment of dental caries, halitosis and periodontitis. It has been also reported that Hamamelis virginiana has anti-bacterial effect on Porphyromonas gingivalis, Fusobacterium nucleatum, Capnocytophaga gingivalis, Veilonella parvula, Eikenella corrodens, Peprostreptococcus micros, and Actinomyces odontolyticus. This study evaluated anti-bacterial effect of Hamamelis virginiana on Streptoccoccus mutans, Haemophylus actinomycetemcomitans, and Klebsiella pneumoniae to expand its anti-bacterial effect on other important oral pathogens and eventually to develop its oral care products or apply to clinical purpose. In this study, anti-bacterial tests for antibiotic disk susceptibility, minimal inhibitory concentration and minimal bactericidal concentration were performed to evaluate anti-bacterial effect of Hamamelis virginiana against Streptoccoccus mutans, Haemophylus actinomycetemcomitans, and Klebsiella pneumoniae. The results showed that Hamamelis virginiana has anti-bacterial effect on all pathogen strains tested in this study and furthermore Hamamelis virginiana possesses bactericidal effect other than bacteriostatic effect on Streptoccoccus mutans, Haemophylus actinomycetemcomitans, Klebsiella pneumoniae. This study indicates that a natural anti-bacterial medication for oral diseases can be developed using Hamamelis virginiana.

• Korean Journal of Microbiology
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• v.52 no.4
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• pp.406-414
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• 2016

Among 6 leu codons, CUG is the most frequently used codon in E. coli. It is recognized by leu-tRNA(CAG) encoded by four genes scattered on two chromosomal loci (leuT and leuPQV ). In the process of constructing a strain with no functional leu-tRNA (CAG) gene on chromosome, we made two mutant strains separately, one on leuPQV locus (${\Delta}leuPQV$), and the other on leuT locus [$leuT^*$(GAG)], where the anticodon of leuT was changed from CAG to GAG, thereby altering its recognition codon from CUG to CUC. We attempted to combine these two mutations by transduction using $leuT^*$(GAG) strain as a donor and ${\Delta}leuPQV$ strain as a recipient. Large and small colonies appeared from this transduction. From PCR and DNA sequencing, large colony was confirmed to be the reciprocal recombinant as expected, but the small colonies contained both mutant $leuT^*$(GAG) and wild type leuT (CAG) genes in the cell. This heterozygous diploid strain did not show any unusual morphology under microscopic observation, but, interestingly, it showed a linear growth curve in rich medium with much slower growth rate than wild type cell. It always formed homogenous small colonies in the selection medium, but, when there was no selection, it readily segregated into $leuT^*$(GAG) and leuT (CAG). From these observations, we suggested that the strain with both $leuT^*$(GAG) and leuT (CAG) genes was not a partial diploid (merodiploid), but a full diploid cell having two different chromosomes. We proposed a model explaining how such a heterozygous diploid cell was formed and how and why its growth showed a linear growth curve.

• Tuberculosis and Respiratory Diseases
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• v.43 no.1
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• pp.14-21
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• 1996

Objectives: Although outbreak of MDR Tb has been a recent problem in western countries, it has been a longstanding problem in Korea. The poor outcome of MDR Tb is mainly due to poor compliance, high rate of side reaction of secondary drugs, and limitation in number of available drugs. Thus, to improve the outcome of MDR Tb, it is crucial to make individualized adequate prescription based on the knowledge of the patterns of resistance to each drugs in the community as well as the natural history. The purpose of present study is to evaluate the clinical features of Korean MDR Tb patients including patterns of drug resistance and success rate of treatment which was prescribed according to the sensitivity tests. Methods: Retroscpective analysis of 71 Korean patients with MDR Tb was made. All strains isolated from patients showed resistence to at least two first line drugs. Patients profile, previous treatment history, patterns of drug resistance, outcome of treatment was analysed. Initial treatment regimen was selected according to the previous treatment history and was modified according to the sensitivity reports. The regimen was composed to include at least 4 sensitive drugs when possible. Results: The patients showed resistance to 4.1 drugs on average. 90% of them were resistant to INH and RFP. Among 71 patients, 35 patients(49%) had cavitary lesions in CXR. Treatment outcome was analysed in 55 patients. 35 patients(67%) were improved after treatment and 18 patients(33%) showed treatment failure. 5 patients showed primary resistance. Treatment outcome could be evaluated in 4 of them and all showed improvement after treatment. 14 patients(20%) had to change their regimens due to drug side effects. The most frequent side effect was elevation of liver enzymes(6 patients). Others included dizziness, hyperuricemia, tinnitus, skin rash, GI troubles. More than 50% of side effects developed within 3 months. In repeated drug sensitivity test, the concordance rate of resistance to INH was 100% and RFP 98%. EMB, PZA showed 80% concordance rate. But in the other drugs, the concordances were less than 50%. Operation was done in 5 patient - 1 patients as a adjunctive means of chemotherapy -. In that case, negative conversion of sputum AFB was done. Conclusion: 2/3 patients of multidrug-resistant tuberculosis were improved by appropriate prescription and regular medication suggesting that more aggressive management and monitoring is indicated in multidrug-resistant tuberculosis.

• Tuberculosis and Respiratory Diseases
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• v.47 no.2
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• pp.184-194
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• 1999

Backgrounds : The advent of penicillin has led to the marked reduction in the mortality from pneumococcal bacteremia, however, the mortality is still relatively high in this post-antibiotic era. Actually the prevalence of infection due to penicillin-resistant penumococci is increasing worldwide, and it is especially high in Korea due to irrelevant use of antibiotics. So, the high mortality of pneumococcal sepsis might be related to the emergence of penicillin-resistant strains, however, many other antibiotics, which eradicate pneumococci effectively, are available in these days. This has led us to suspect the role of penicillin-resistance in the high mortality rate. In this study, we evaluated the effect of penicillin resistance on the mortality of patients with penumococcal bacte remia. Methods: The study population consisted of 50 adult patients with penumococcal bacteremia who were admitted between Jan, 1990 and July, 1997. Medical records were analyzed retrospectively. Results: Most of the patients (96%) had underlying diseases. The most common local disease associated with pneumococcal bacteremia was pneumonia (42%), which was followed by spontaneous bacterial peritonitis (14%), cholangitis (10%), meningitis (8%), liver abscess (4%), pharyngotonsillitis (4%), sinusitis (2%) and cellulitis (2%). While the overall case-fatality rate in this study was 24%, it was higher when peumococcal bacteremia was associated with pneumonia (42%) or meningitis (50%). The rate of penicillin resistance was 40%, which was increased rapidly from 1991. The rate of penicillin resistance was significantly higher in patients with the history of recent antibiotics use and hospitalization within 3 months respectively. The clinical manifestations, that is, age, severity of underlying diseases, nosocomial infection, associated local diseases, and the presence of shock or acute renal failure were not statistically different between the patients with penicillin-resistant and -sensitive pneumococcal bacteremia. The mortality of patients infected with penicillin-resistant pneumococci was not statistically different from those with penicillin-sensitive pneumococcal bacteremia. Conclusion: Penicillin resistance is not associated with high mortality in adult patients with pneumococcal bacteremia. As the overall mortality is high, active penumococcal vaccination is recommended in patients with high risk of infection.

• Tuberculosis and Respiratory Diseases
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• v.60 no.2
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• pp.171-179
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• 2006

Background : Despite the emerging danger of MDR-TB to human beings, there have only been a limited number of drugs developed to treat MDR-TB since 1970. This study investigated the cross-resistance rate between rifampicin (RFP) and rifabutin (RBU) in order to determine the efficacy of rifabutin in treating MDR-TB. In addition, the results of rifabutin were correlated with the rpoB mutations, which are believed to be markers for MDR-TB and RFP resistance. Methods : The MICs of RBU were tested against 126 clinical isolates of MDR-TB submitted to the clinical laboratory of National Masan TB Hospital in 2004. Five different concentrations ($10-160{\mu}g/ml$) were used for the MICs. The detection of the rpoB mutations was performed using a RFP resistance detection kit with a line probe assay(LiPA), which contains the oligonucleotide probes for 5 wide type and 3 specific mutations (513CCA, 516GTC, and 531TTG) The rpoB mutation was determined by direct sequencing. Results : The rate of cross-resistance between RFP and RBU was 70.5%(74/105) at $20{\mu}g/ml$ RBU(ed note: How much RFP?) Most mutations (86.3%) occurred in the 524~534 codons. The His526Gln, His526Leu, Leu533Pro, Gln513Glu, and Leu511Pro mutations(Ed note: Is this correct?) were associated with the susceptibilty to RBU. Conclusion : Based on the cross-resistance rate between RFP and RBU, RBU may be used effectively in some MDR-TB patients. Therefore, a conventional drug susceptibility test for RBU and a determination of the critical concentration are needed. However, rpoB gene mutation test may be have limited clinical applications in detecting RBU resistance.

• Tuberculosis and Respiratory Diseases
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• v.65 no.2
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• pp.91-98
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• 2008

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is the most common organism associated with nosocomial infections. MRSA infections are becoming increasing important because they have emerged no only as healthcare-associated (HA) infections but also as community-associated (CA) ones. This study examined the moleculo-epidemiology of MRSA, which was isolated from nasal swabs in the intensive care unit (ICU) at Konyang University Hospital. MRSA are classified into HA-MRSA and CA-MRSA. Methods: From June to September 2006, 353 patients who were admitted to the ICU in Konyang University Hospital were enrolled in this study. Single nasal swabs were obtained for culture in the ICU on the 1st day. Pulsed-field gel electrophoresis and the antimicrobial resistant patterns were analyzed between HA- and CA-MRSA. An antimicrobial sensitivity test was also performed. Results: Forty two strains of MRSA were isolated from 353 patients (11.9%). Among the 42 isolates, HA-MRSA and CA-MRSA were found in 33 (78.6%), and 9 (21.4%), respectively. Eleven different PFGE types (type A to K) were identified. Types A (n=9) and B (n=7) were the most common for HA-MRSA, and types A (n=2) and B (n=2) were identified in CA-MRSA. The proportion of types A and B in CA-MRSA (44.4%) was similar to that in HA-MRSA (48.5%). The rates of resistance rates to erythromycin and ciprofloxacin were higher in HA-MRSA than in CA-MRSA. Conclusion: The rate of isolation of MRSA in an ICU setting was 11.9%. HA-MRSA was isolated more frequently than CA-MRSA. The rate of resistance of HA-MRSA to erythromycin and ciprofloxacin was higher than that of CA-MRSA. Despite the small number of subjects, the main isolates (type A and B) of CA-MRSA were similar to those of HA-MRSA.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.5
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• pp.577-583
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• 2012

This study investigated the antioxidant activities, and antimicrobial activity $in$ $vitro$ of an 80% ethanol extract from garlic, daikon, leek, ginger, onion, and green onion, which are widely-used ingredients in Korean food that contain sulfur. The total polyphenol content in ginger and leek extracts showed a high value ($233.63{\pm}4.59$ and $220.98{\pm}10.56$ mg/g GAE) and onions, leeks, garlic, and daikon followed by with $69.07{\pm}1.42$, $68.83{\pm}2.11$, $19.41{\pm}0.40$, $19.05{\pm}03.32$ mg/g GAE, respectively. DPPH radical scavenging activity was highest with ginger extracts ($1.57{\pm}0.15$ mg/mL as $IC_{50}$) followed in order of decreasing activity by leeks, onions, daikon, green onions, and garlic. The results of ABTS radical scavenging activity and FRAP value showed higher antioxidant activity in extracts from ginger and leek. The order of vegetables with most to least prevalent ABTS radical scavenging activity was green onions, onions, garlic, and finally daikon. From greatest to least FRAP value, the relevant vegetables were green onions, onions, daikon, and garlic (p<0.05). Ginger extracts showed promise against seven strains of microbes: $Bacillus$ $cereus$, $Bacillus$ $subtillis$, $Staphylococcus$ $aureus$, $Lactobacillus$ $plantarum$, $Escherichia$ $coli$, $Salmonella$ $enterica$, and $Pseudomonas$ $aeruginosa$. Garlic extracts (5 mg/disc) showed strong antimicrobial activity against $B.$ $cereus$ (22.3 mm) and $E.$ $coli$ (24.3 mm). Extracts of both onion and green onion showed antimicrobial activity against only $E.$ $coli$ (12.7 and 10.3 mm) and $B.$ $cereus$ (12.0 and 12.5 mm) at 10 mg/disc, and the inhibition zone diameter from extracts of garlic and leeks were 18.0 mm and 10.4 mm vs. $L.$ $plantarum$ at 10 mg/disc. This study showed positive antioxidant activities for ginger and leeks, and positive antimicrobial activities for leeks and garlic. These sulfur-containing vegetables are widely used in Korean food. Leeks especially could serve as a functional food preservative.

• KSBB Journal
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• v.25 no.2
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• pp.155-166
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• 2010

Studies on the production of cell-wall bound innerpolysaccharides (IPS) (soluble ${\beta}$-D-glucan) have been performed by use of suspended myelial cultures of Inonotus obliquus. This product has promising potentials as an effective antidiabetic as well as an immunostimulating agents. As a first step to enhanced production of IPS, Intensive strain improvement programs were carried out by obtaining a large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because about fivefold higher amount of protoplasts ($2.3{\times}10^6$ protoplasts/mL) could be recovered with relatively high regeneration rates of $10^{-2}{\sim}10^{-3}$ by applying a modified filtration method, as compared to the previously used trapping method. A basic protocol necessary for UV-mutation of the protoplasts was also developed, resulting in several overproducing variants with good fermentation properties. Since the amount of IPS extracted from the mycelial cell walls of I. obliquus turned out to be almost constant per g DCW, increase in cell mass was considered the most important factor for the enhancement in IPS production. Therefore, attempts were made to screen mutant cells showing rapid mycelial growth rate in the final suspended cultures. Notably, the mutant strains showing an active cellgrowth in the preceding solid growth cultures were observed to produce higher amount of IPS in the suspended fermentations as well. A striking mutant, OBLQ756-15-5 strain, obtained from the survivors of a harsh UV-treated condition (97% death rate) was found to stably produce as high cell mass as 22 g DCW/L in the final fermentations. Currently, this strain is being tested for development of a scaled-up fermentation process for mass production of IPS.

• KSBB Journal
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• v.22 no.5
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• pp.305-312
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• 2007

It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.