• Korean Journal of Microbiology
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• v.39 no.4
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• pp.223-229
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• 2003

The purpose of this work was to perform the characterization of NAD(P)H-nitroreductase isolated from Stenotrophomonas sp. OK-5 capable of degrading 2,4,6-trinitrotoluene (TNT). Initially, NADP(H)-nitroreductase by a series of purification processes including ammonium sulfate precipitation, DEAE-sepharose, andQ-sepharose was prepared. From samples harvested from fraction collector, three different fractions (I, II & III)having the enzyme activity of NAD(P)H-itroreductase were detected. Specific activities of three fractions I, II,and III of NAD(P)H-nitroreductase were determined to approximately 5.06 unit/mg, 4.95 unit/mg and 4.86 unit/mg, and concentrated to 10.5, 9.8, and 8.9-fold compared to crude extract, respectively. Among these three fractions,the fraction I of NAD(P)H-nitroreductase demonstrated the highest specific activity in this experiment. Several factors affecting on the enzyme activity of NAD(P)H-nitroreductase (fractions I, II & III) were investigated.The optimum temperature of all NAD(P)H-nitroreductase (fractions I, II & III) was 30oC, and the optimal pH was approximately 7.5. Metal ions such as Ag+, Cu2+, Hg2+ inhibited approximately 80% enzyme activity of all NAD(P)H-nitroreductase, and the enzyme activities were decreased about 30-40% inhibition in the presence of Mn2+ or Ca2+. However, Fe3+ showed stimulatory effect on the enzyme activity. The molecular weights of NAD(P)H-nitroreductase (fractions I, II & III) were measured about 27 kDa on the SDS-PAGE.

• Korean Journal of Microbiology
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• v.51 no.1
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• pp.1-6
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• 2015

The term "P2 sir-associated helper inefficiency" has been used to define the inefficient helper capability of P2 sir mutants for their satellite bacteriophage P4. The aim of this study was to investigate the factors overcoming P2 sir-associated helper inefficiency. At first, we verified whether the P2 cos region containing P4 sid71 cosP2 could overcome P2 sir-associated helper inefficiency with P2 sir3. The result was that P4 sid71 cosP2 could not overcome P2 sir-associated helper inefficiency with P2 sir3. Instead of cos region of P2, the size of the DNA packaged into a $P2_{sir}$-sized head seems to be important for overcoming P2 sir-associated helper inefficiency. In the present work, three kinds of P4 derivatives with packaged DNA sizes between those of P4 ost1 and P4 ost2, were constructed through DNA manipulation. In one P4 derivative, P4 sid71 delRI::apr, the size of the packaged DNA was identified with a CsCl buoyant equilibrium density gradient experiment. According to the burst sizes of the P4 derivatives, they could overcome P2 sir3-associated helper inefficiency. The size of the P4 derivative DNA suitable for packaging into a $P2_{sir3}$-sized head was 28-29 kb.

• Journal of Microbiology
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• v.45 no.1
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• pp.21-28
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• 2007

In order to search for specific genotypes related to this unique phenotype, we used whole genomic DNA microarray to characterize the genomic diversity of Helicobacter pylori (H. pylori) strains isolated from clinical patients in China. The open reading frame (ORF) fragments on our microarray were generated by PCR using gene-specific primers. Genomic DNA of H. pylori 26695 and J99 were used as templates. Thirty-four H. pylori isolates were obtained from patients in Shanghai. Results were judged based on In(x) transformed and normalized Cy3/Cy5 ratios. Our microarray included 1882 DNA fragments corresponding to 1636 ORFs of both sequenced H. pylori strains. Cluster analysis, revealed two diverse regions in the H. pylori genome that were not present in other isolates. Among the 1636 genes, 1091 (66.7%) were common to all H. pylori strains, representing the functional core of the genome. Most of the genes found in the H. pylori functional core were responsible for metabolism, cellular processes, transcription and biosynthesis of amino acids, functions that are essential to H. pylori's growth and colonization in its host. In contrast, 522 (31.9%) genes were strain-specific genes that were missing from at least one strain of H. pylori. Strain-specific genes primarily included restriction modification system components, transposase genes, hypothetical proteins and outer membrane proteins. These strain-specific genes may aid the bacteria under specific circumstances during their long-term infection in genetically diverse hosts. Our results suggest 34 H. pylori clinical strains have extensive genomic diversity. Core genes and strain-specific genes both play essential roles in H. pylori propagation and pathogenesis. Our microarray experiment may help select relatively significant genes for further research on the pathogenicity of H. pylori and development of a vaccine for H. pylori.

• Korean Journal of Food Science and Technology
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• v.22 no.6
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• pp.611-617
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• 1990

This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

• Journal of the korean academy of Pediatric Dentistry
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• v.39 no.1
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• pp.26-33
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• 2012

The purpose of this study was to find out the effect of dentifrice containing Horseradish (Armoracia rustica) root extracts on improvement of oral hygiene. The clinical evaluation was done participated by 80 adults volunteers who provided a informed consent for their participation. The participants were divided into two groups ; control group (using toothpaste without Horseradish extracts) and experimental group (using toothpaste containing Horseradish extracts). The checklist of the experiment includes the plaque index, gingival index and anti-caries activity. The anti-caries activity was measured by cultivating Streptococcus mutans for 48 hours then estimating the number of increased bacteria using Dentocult SM. The double-blind method was used in this study. The results of this study can be summarized as follows; 1. The plaque formation decreased 28% in control group, 58% in experimental group using dentifrice with Horseradish extracts compared to baseline data. 2. For gingival index, control group and experimental group showed 26% and 40% decrease compared to baseline data, respectively. 3. The score of caries activity test decreased by 4% for control group and 69% for experimental group compared to baseline data. According to the results, a dentifrice containing Horseradish extracts can improve oral hygiene.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.152-156
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• 2010

Disposable contact lenses, which are one type of soft contact lenses, provide convenience in use, but also cause various ocular infectious diseases. Microorganisms that cause eye diseases include Acanthamoeba, bacteria, Fungi, and so on. It is impossible to prevent microorganism contamination completely due to the use of hands as wearing contact lenses. The contamination by various microorganisms leads to infectious keratitis, but it is not well known for the exact microorganisms that affect the disease. For this reason, to identify the microorganisms, two groups that are commonly used for disinfection of lenses were divided: normal saline solution and multiple purpose solution. Using these solutions the degree of microorganism contamination was observed according to the days of 1, 3, 5, 10, and 15. Twenty students by two groups from Ophthalmic Optics department at D college in Daegu Metropolitan city participated in the experiment after their ocular health conditions were checked. During they wore one-day disposable lenses for 1, 3, 5, 10, and 15 days, bacteria were cultured in media. The results, which were Gram stained by selecting the cultured colonies, show as followings: Gram positive cocci 33%, Gram-negative cocci 2%, Gram positive bacilli 34%, and the Gram negative bacilli 31%, respectively. As for the identification of potential pathogens, VITEK system and API kit methods were used. Keratitis caused by bacteria known as Staphylococcus aureus, Pseudomonas aeruginosa were detected as a result of wearing contact lenses. This study examined the distribution of bacteria as wearing one-day disposable contact lenses and pathogenic bacteria according to the duration of wearing them. In conclusion, the importance of hygiene when using contact lenses is suggested.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.120-124
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• 2016

A certain size of DNA (28-29 kb long) to be packaged into P2-size head and the mutation in sid gene of bacteriophage P4 are the major factors to overcome "P2 sir-associated helper inefficiency". To clarify whether the presence of sid mutation is essential to overcome "P2 sir-associated helper inefficiency" or not, we tested the P4 derivative, P4 delRI::kmr, which is $sid^+$ and whose genome size supposed to be 28.5 kb long in the case of being packaged into $P2_{sir3}$-sized large head. As P4 delRI::kmr showed the low EOP with P2 sir3 lysogen, P4 delRI::kmr phage stock was prepared in P2 sir3 lysogen host to increase the EOP with P2 sir3 lysogen. Through this process, P4 delRI::kmr had been adapted for P2 sir3 lysogen. With a CsCl buoyant equilibrium density gradient experiment and gel electrophoresis of the isolated DNA, it was evident that the adaptation of P4 delRI::kmr for P2 sir3 lysogen was caused by the conformational change of DNA to be packaged into large head. The burst size determination experiments with P4 delRI::kmr phage stock adapted for P2 sir3 lysogen and normal P4 delRI::kmr phage stock showed that not the sid mutation but the size of DNA to be packaged (28-29 kb long) was essential to overcome "P2 sir-associated helper inefficiency".

• Korean Journal of Microbiology
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• v.36 no.2
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• pp.91-96
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• 2000

The cultural and physiological conditions for the T-2 toxin [4,15-diacetoxy-8-(3-mety1butyloxy)-12,13- epoxy-trichothec-9-en-3-01, $C_{24}H_{30}O_9$] production by Fusarium spp. were studied. Thin layer chromatography (TLC) assay and the microbiological assay uslng Rhodotomla rubra were used to quantitate tbe T- 2 toxin. Among the four strains of Fusarium spp., F tn'cinctum NRRL 3299 was best for T-2 toxin production. In solid culture, white com grit medium was best for T-2 toxm production. Temperature played a critical role in the production of T-2 toxin. T-2 toxin production was favored by long duration of low-temperature incubation. The growth and toxin production were relatively high on galactose, fructose, glucose, and sucrose media, when each was used as a sole carbon source, and relatively low on sorbitol, glycerol, and lactose media. For nitrogen sources, $NH_4^(+) and NO_3^{-}were used well as a sole nitrogen source, but $NO_2^-$ was not used. Initial pH and speed of shaker also affected the production of T-2 toxin. From temperature shifting experiment, it is clear that T-2 toxin metabolic pathway is regulated by temperature-dependent enzyme depression or enzyme induction system.

• Korean Journal of Microbiology
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• v.36 no.4
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• pp.279-284
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• 2000

Simultaneous reduction of Cr(VI) and degradation of phenol was observed in batch and bench-scale continuous stirred tank reactors using Rhodococcus sp. CP01 isolated from leachate. The strain CP01, which was capable of utilizing phenol as a sole source of carbon and energy, completely reduced added hexavalent chromium (0.25 mM) to its trivalent form during 60 hr batch assay under optimal conditions (pH 7.0 and 1,000 mg/L of phenol concentration). The rates of Cr(VI) reduction and phenol degradation were estimated as 4.17 $\mu$M Cr(VI) and 38.4 mg phenol.$L^{-1}{\cdot}hr^{-1}$, respectively. The continuous culture experiment was conducted for 46 days using synthetic feed containing different levels of chromate (0.0625 to 0.25 mM) and phenol(1,000 to 4,000 mg/L). With a hydraulic retention time of 100 hr, Cr(VI) reduction efficiency was mostly 100% for influent Cr(VI) and phenol concentrations of 0.125 mM and 3,000 mg/L, respectively. During quasi-steady-state operation, specific rate of Cr(VI) reduction was calculated as 0.34 mg Cr(VI).g $protein^{-1}{\cdot}hr^{-1}$ which was comparable to reported values obtained by using glucose as growth substrate. The results suggest the potential application of biological treatment for detoxification of wastewater contaminated simultaneously with Cr(VI) and pheonol.

• Korean Journal of Microbiology
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• v.26 no.4
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• pp.283-290
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• 1988

A spontaneous revertant of mutation rbsB103 that is ribose taxis-positive was characterized. This revertant was found to be export-competent in the export of ribose-binding protein shown by the disappearance of accumulated mutant precursor protein and the export of mature ribose-binding protein to the periplasm. The reversional change was shown to be in the region of risB gene that codes for the amino terminal portion of ribose-binding protein. Analysis by high-performance liquid chromatography of peptide patterns of ribose-binding proteins confirmed the relationship between the wild-type and the revertant proteins as shown for the mutant previously (Iida et al., 1985). When the processing rate of presursor proteins from the wild type and the revertant strain in vivo was compared by pulse-chase experiment, it was found that processing is less efficient than normal in the revertant. Purified mature proteins from both wild-type and revertant were subjected to amino acid sequencing. The results confirmed the amino acid changes deduced from the DNA sequencing and showed that processing of the revertant precursor occured in the correct position even though there are two different amino acids present in the signal sequence.