• Korean Journal of Microbiology
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• v.51 no.3
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• pp.280-287
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• 2015

Probiotics are microbial food supplements or components of bacteria which have traditionally been added to dairy foods for extra health boost. Our aim was to evaluate the hepatoprotective effect of Bifidobacterium adolescentis SPM0212 as probiotics, which we previously found has potential anti-hepatitis B virus activity. The study was conducted using Wistar albino rats and probiotics were treated orally for 9 days consecutively and acute liver injury was induced by administration of carbon tetrachloride ($CCl_4$) on the 7th and 8th days. Liver damage was assessed by quantifying serum activities of glutamate oxaloacetate transaminase (SGOT) and glutamate pyruvate transaminase (SGPT), as well as by histopathological examination. B. adolescentis SPM0212 significantly prevented the elevation of SGOT and SGPT levels, and reduced the negative effect of $CCl_4$ on body and organ weights. Histopathological study revealed the livers of the carbon tetrachloride treated rats showed almost complete loss of normal hepatocyte architecture, but that rats treated with B. adolescentis SPM0212 showed minimal damage and normal hepatocyte architecture. Our results suggest that B. adolescentis SPM0212 be considered useful probiotics for protecting the liver from xenobiotics and hepatitis B virus, and as well as useful as a functional food for maintaining human health.

• Korean Journal of Microbiology
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• v.49 no.4
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• pp.369-376
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• 2013

We isolated the ${\beta}$-glucosidase producing bacteria (BGB) in ginseng root system (rhizosphere soil, rhizoplane, inside of root). Phylogenetic analysis of the 28 BGB based on the 16S rRNA gene sequences, BGB from rhizosphere soil belong to genus Stenotrophomonas (3 strains), Bacillus (1 strain), and Pseudoxanthomonas (1 strain). BGB isolates from rhizoplane were Stenotrophomonas (16 strains), Streptomyces (1 strain) and Microbacterium (1 strain). BGB from inside of root were categorized into Stenotrophomonas (3 strains) and Lysobacter (2 strains). Especially, Stenotrophomonas comprised the largest portion (approximately 90%) of total isolates and Stenotrophomonas was a dominant group of the ${\beta}$-glucosidase producing bacteria. We selected strain 4KR4, which had high ${\beta}$-glucosidase activity (108.17 unit), could transform ginsenoside Rb1 into Rd, Rg3, and Rh2 ginsenosides. In determining its relationship on the basis of 16S rRNA sequence, 4KR4 strain was most closely related to Stenotrophomonas rhizophila e-$p10^T$ (AJ293463) (99.62%). Therefore, on the basis of these polyphasic taxonomic evidence, the ginsenoside Rb1 converting bacteria 4KR4 was identified as Stenotrophomonas sp. 4KR4 (=KACC 17635).

• Korean Journal of Microbiology
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• v.34 no.1_2
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• pp.20-25
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• 1998

To obtain higher yield of 2,3-dihydroxybiphenyl(2,3-DHBP) dioxygenase by recombinant E. coli CK1092 carrying pcbC gene of Pseudomonas sp. P20, the environmental and physiological factors were investigated and the cultural conditions using jar fermentor were studied. E. coli CKl092 was grown in LB medium supplemented with 2% sucrose, as a basal medium. The effect of various metal ions on the enzyme production was investigated. In particular, the enzyme production increased in the presence of $Fe^{3+}$ and $Fe^{2+}$, and showed the maxium at the concentration of $10^{-5}M$. The enzyme production was increased by 55% in the medium containing $Fe^{3+}$ ($10^{-5}M$) ion. The optimal temperature and initial pH for cell growth were $37^{\circ}C$ and 7.0, respectively. In the culture using a fermentor at $37^{\circ}C$, the optimal conditions for the enzyme production were obtained at the initial pH 7.0, 1 v/v/m of aeration rate, 200 rpm of agitation speed. It was found that enzyme activity was higher when cultivated without pH control than with pH control.

• Journal of Veterinary Clinics
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• v.6 no.2
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• pp.307-318
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• 1989

In recent years much attention has been paid to swine respiratory infection caused by Pasteurella(P) multocida with rapid expansion of pork Industry in Korea. The present study was performed to observe the etiologic situation of P. multocida infection by bacteriological, serological(serotyping) and pathological examinations with the lungs respectively. In addition antibiotic susceptibility test was carried out against the isolated strains of P. multocida. The results obtained are as follows : 1. Eighteen strains(12.8％) wert isolated from the 140 cases of swine lungs examined, and biological and biochemical characteristics of the isolates were the sam as those in the references of other workers, whereas some differences were observed in sugar fermentation and enzyme activity according to the strain of isolates. 2. Capsular serotyping performed on 18 P. multocida revealed that 13 strains(72.2％) were A type and 5 strains(27.8％) were D type, respectively. 3. When serotyping was performed against somatic antigen on 18 strains capsular types of which were identified as described above 9(50％), 3(16.7％) and 4(22.2％) strains belong to 1:A, 3:A and 2:D, respectively, but untypable 2 strains(11.1％) were observed. 4. Antibiotic susceptibility test by employing disc method for 24 kinds of drugs revealed that 15 kinds of antibiotics were sensitive to 18 strains of P. multocida isolated such as ampicillin(l00％), penicillin(100％), cloxacillin(56％), piperacillin(70％), cefotaxime(30%), minocycline(60％), chloramphenicol(95％), erythromycin(39％), kanamycin(17％), gentamicin(70%), amikasin(30%), colistin(78％) and nalidixic acid(5％), respectively, but resistant to 9 kinds of antibiotics such as sulpenicillin, cefazolin, cephalothin, cefametazol, cefoperazone, kitasamycin, oleandomycin. lincomycin and bacitracin. 5. Pathological features of 60 cases of swine lungs indicated that pneumonic .lesions were observed in 38 cases(63.3％) examined by macroscopic finding, in which lesions of 8 cases(13.4％) would correspond to those of mycoplasmal infection, and 30 cases(50％) were similar to viral infection by histopathological finding, whereas 22 cases(36.7％) were considered to be normal by ecropsy or histopathological finding.

• Korean Journal of Veterinary Research
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• v.34 no.3
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• pp.517-527
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• 1994

An attempt was made to isolate a causative viral agents from the fecal specimens of the diseased dogs with the gastroenteritis symptoms. Two coronavirus-like agents were isolated by serial dilution end point method and plaque assay. The isolates were characterized in terms of cytopathology, antigenicity, replication, physicochemical and morphological properties. The results obtained through the experiment were as follows; 1. Among 7 fecal specimens collected from the dogs with enteric disease, 2(28.6%)coronavirus-like agents showing typical cytopathic effects of canine coronavirus were isolated, and designated as CCV D1 and CCV D2, respectively. 2. By the cross-neutralization test and indirect immunofluoresence antibody test, the isolates were antigenically indentified as the standard CCV. The viruses were replicated only in the cytoplasm of A-72 cells. 3. The isolates showed no haemagglutinating activity against the erythrocytes from 11 kinds of animals. 4. The electron microscopic observation for the isolates showed spherical and pleomorphic features, covered with club-shaped projections on the surface. The size of particles was ranged from 70 to 150nm. 5. In one-step growth curve for the isolates in A-72 cells, maximum titers of intracellular vius was $10^{4.6}$ $TCID_{50}/0.1ml$ at 46 hrs postinoculation(pi) of CCV Dl and $10^{4.4}$ $TCID_{50}/0.1ml$ at 34 hrs pi of CCV D2. The maximum titers of extracellular virus was $10^{5.5}$ $TCID_{50}/0.1ml$ at 58 hrs pi of CCV D1 and $10^{5.8}$ $TCID_{50}/0.1ml$ at 46 hrs pi of CCV D2. 6. In physicochemical property test, the isolates were very sensitive to choroform and were found to be RNA virus. The viruses was stable at pH 3.0 for 1 hr and at $22{^{\circ}C}$ for 5 hrs. However, infectivity titers reduced remarkably by treatment with $56{^{\circ}C}$ for 10min.

• Korean Journal of Microbiology
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• v.33 no.1
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• pp.1-6
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• 1997

5-Enolpyruvylshikimate 3-phosphate(EPSP) synthase is the sixth enzyme of the shikimate pathway that synthesizes aromatic amino acids. The enzyme is a primary target for the glyphos'lte which is a broad-spectrum and environmetally safe herbicide. As a first step toward development of glyphpsate-resistant EPSP synthase, the EPSP synthase gene(aroA) was amplified by polymerase chain reaction and cluned into pET-25b vector. In this construct. designated pET-aro, the aroA gene is expressed under control of strong T7 promoter. and the EPSP synthase is produced as a fusion protein with pelB leader at N-terminus and HSV-tag and His-tag at C-terminus. When the pET-aro clone was induced to produce the enzyme, it was found that the EPSP synthase was successfully exported to peri plasmic space. The periplasmic transport was greatly dependent on the induction temperatures. Among the induction temperatures examined($25^{\circ}C$, $30^{\circ}C$, $34^{\circ}C$ and $37^{\circ}C$). induction at $34^{\circ}C$ gave rise to maximal periplasmic transport. The recomhinant EPSP synthase could have been purified hy $Ni^{2+}$ -affinity chromatography using the His-tag. and detected hy anti-HSV -tag antibody. The recombinant EPSP synthase also hound to phosphocellulose resin and was eluted hy shikimate 3-phosphate and phosphoenolpyruvate. as expected. The recombinant EPSP synthase purified from phosphocellulose resin showed typical EPSP synthase activity.

• Korean Journal of Microbiology
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• v.32 no.3
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• pp.222-231
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• 1994

Kinetic analysis was done to elucidate the reaction mechanism of purine nucleoside phosphorylase (PNP) in Saccharomyces cerevisiae. The binary complexes of PNP${\cdot}$phosphate and PNP${\cdot}$ribose 1-phosphate were involved in the reaction mechanism. The initial velocity and product inhibition studies demonstrated were consistent with the predominant mechanism of the reaction being an ordered bi, bi reaction. The phosphate bound to the enzyme first, followed by nucleoside and base were the first product to leave, followed by ribose 1-phosphate. The kinetically suggested mechanism of PNP in S. cerevisiae was in agreement with the results of protection studies against the inactivation of the enzyme by sulfhydryl reagents, p-chloromercuribenzoate (PCMB) and 5,5'-dithiobisnitrobenzoate (DTNB). PNP was protected by ribose 1-phosphate and phosphate, but not by nucleoside or base, supporting the reaction order of ordered bi, bi mechanism. PCMB or DTNB-inactivated PNP was totally reactivated by dithiothreitol (DTT) and the activity was returned to the level of 77% by 2-mercaptoethanol, indicating that inactivation was reversible. The kinetic behavior of the PCMB-inactivated enzyme had been changed with higher $K_m$ value of inosine and lower $V_m$, and was restored by DTT. Inactivation of enzyme by DTNB showed similar pattern of K sub(m) value with that by PCMB, but had not changed the $V_m$ value, significantly. Negative cooperativity was not found with PCMB or DTNB treated PNP at high concentration of phosphate.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.176-180
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• 1989

There are three pseudourdine ($Psi$)bases in the E. coli $tRNA^{phe}$ In order to study the function of the pseudouridine bases in the $tRNA^{phe}$, changes of bases $tRNA^{phe}$ gene to other bases were undertaken by the site-specific mutagenesis. Site-specific mutagenesis of T in the pheW gene, a $tRNA^{phe}$ gene of E. coli, corresponding to the baseat the No.32 position to C and also T corresponding to the base at the No.39 position to C were performed using Kunkel's uracil-containing template method. Identification of mutants were undertaken by the KNA sequencing techniques of the mutated pheW genes and activities of the mutated pheW genes complementing to E. coli NP37 mutant($pheS^{-ts}$) using the recombinant plasmid containing the mutated genes. Neither NP37 harboring pheW gene mutated at No.32 position nor NP37 harboring pheW gene mutated at No.39 position can be grown at non-permissive temperature. The result means that both mutated pheW genes can not complement to E. coli NP37, and that the pseudouridine bases are essential to the activity of the E. coli $tRNA^{phe}$ in vivo.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.120-125
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• 1998

A plasmid pNPG contains a genomic DNA complementing npgA1 which is located on the left arm of linkage group I. It transformed Aspergillus nidulans at a high frequency. No abortive transformants were observed and the $Trp^+$ transformants were all $Npg^+$. The 10.4 kb Psti fragment of the genomic DNA was subcloned into pILJ16, which increased the transformation efficiency by more than 200-folds. The transformants were mitotically unstable and yielded $Arg^-$ conidia at the frequency of more than 80%. An additional gene cloned into the plasmid containing the fragment was always lost with $argB^+$ marker. These characteristics strongly indicate the possibility that the plasmids autonomously replicate. The full activity of enhanced transformation was retained on the 4.9 kb EcoRI-HaeIII fragment. The DNA segment was similar to AMA1 rather than ANS1 in function and designated AMA2.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.73-79
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• 2010

The Korea Polar Research Institute (KOPRI) has assembled a culture collection of cold-adapted bacterial strains from both the Arctic and Antarctic. To identify excellent protease-producers among the proteolytic bacterial collection (874 strains), 78 strains were selected in advance according to their relative activities and were subsequently re-examined for their extracellular protease activity on $0.1{\times}$ ZoBell plates supplemented with 1% skim milk at various temperatures. This rapid and direct screening method permitted the selection of a small group of 15 cold-adapted bacterial strains, belonging to either the genus Pseudoalteromonas (13 strains) or Flavobacterium (2 strains), that showed proteolytic activities at temperatures ranging between $5-15^{\circ}C$. The cold-active proteases from these strains were classified into four categories (serine protease, aspartic protease, cysteine protease, and metalloprotease) according to the extent of enzymatic inhibition by a class-specific protease inhibitor. Since highly active and/or cold-adapted proteases have the potential for industrial or commercial enzyme development, the protease-producing bacteria selected in this work will be studied as a valuable natural source of new proteases. Our results also highlight the relevance of the Antarctic for the isolation of protease-producing bacteria active at low temperatures.