• Food Science of Animal Resources
• /
• v.21 no.2
• /
• pp.133-141
• /
• 2001

Adherence of probiotic bacteria to intestinal epithelium is found to be the most principal characteristics among the various physiological functionality. This study was conducted to investigate the effect of bifidobacterial growth properties and condition on the Caco-2 cell adherence and to construct a basic data on adherence-related research. Among 20 strains of bifidobacteris tested, when measured by cell surface hydrophobicity(CSH) and cell agglutination(CA), Bifidobacterium bifidum ATCC29521, Bif. adolescentis K8, and Bif. infantis K9 were selected. Using these strains, variations of Caso-2 cell adherence depending upon experimental condition were analyzed. The results obtained are as follows : Even though Bif. bifidum ATCC29521, Bif. adolescentis K8, and Bif. infantis K9 reached more 85% cell surface hydrophobicity there was no significant difference in cell agglutination, when reached 31.54$\pm$0.54mg/ml. By direct count method for adherence, viable cell count of M3, K1, K2, K8, K9 and K10 reached more 100 counts per 100 Caco-2 cells. When Bif. bifidum ATCC29521, Bif. adolescentistis K8, and Bif. infantis K9 were used to compare the adherence depending upon viable cell counts, reaction time, and growth phase, the more viable cell count, and the more adhered cell counts, the less adherence percentage. In addition, there was no difference in adherence percentage of bifidobacteria when bifidobacteria was incubated from 1 to 8 hrs after Caco-2 cells already formed monolayer. Considering of the effect of growth phase of bifidobacteria on adherence variation, all strains showed the highest adherence during the early stage of stationary phase. In conclusion, adherence of bifidobacteria was affected by strain specificity, viable cell count, and growth activity.

• Journal of Microbiology and Biotechnology
• /
• v.1 no.2
• /
• pp.116-120
• /
• 1991

The transformant Bacillus subtilis ED213 carrying the pSO100 which cloned the cdd gene encoding cytidine deaminase (cytidine /2'-deoxycytidine aminohydrolase, EC 3.5.4.5, CDase) originated from wild type B. subtilis was cultivated in Spizizen minimal medium (SMM). To overcome poor expression of the cdd gene in SMM medium, the medium compositions and growth conditions were optimized. The optimized medium compositions and growth conditions were cytidine concentration of 80 mg/l, glycerol of 25 g/l, and $(NH_4)_2SO_4$ of 10 g/l, along with $37^{\circ}C$ and pH 7.0. The intracellular CDase production was increased 3 times from 1,000 unit/ml to 3,200 unit/ml, and extracellular CDase also increased from nearly undetectable amounts to 1,500 unit/ml. The cytidine concentration was signified as the most critical compositional factor for overproduction of CDase by increasing the cell density mainly in culture broth. The plasmids were more stable in cells that were grown in original SMM medium with stability of 90% compared to those grown in optimized SMM medium with stability of 80% after 48 hours cultivation. The most active amplification of plasmid was occurred in the logarithmic phase, which showed a value around four times higher than the initial copy number. In the exponential phase, the CDase production was closely related to the plasmid copy number along with the cell density. However, it was not accorded with cell density at the stationary phase.

• Archives of Pharmacal Research
• /
• v.23 no.2
• /
• pp.178-181
• /
• 2000

In order to study the simultaneous determination of (+)- and (-)-cetirizine in human urine we have developed a chiral separation method by HPLC. A chiral stationary phase of $\alpha$$_1$-acidglycoprotein, the AGP-CSP was used to separate the enantiomers. The pH of the phosphate buffer, as well as the content of the organic modifier in the mobile phase, markedly affected the chromatographic separation of (+)- and (-)-cetirizine. A mobile phase of 10 m㏖/1 phosphate buffer (pH 7.0)-acetonitrile (95 : 5, v/v) was used for the urine assays. Ultraviolet absorption was monitored at 230nm and roxatidine was employed as the internal standard for quantification. (+)-Cetirizine, (-)-cetirizine and the internal standard were eluted at retention times of 12, 16, and 32 mins, respectively. The detection limit for cetirizine enantiomers was 400 ng/$m\ell$ of urine. A pharmacokinetic study was conducted with the help of 5 healthy female volunteers who were administered with a single oral dose of racemic cetirizine (20 mg). The peak area ratios provided by the cetirizine enantiomers were linear(r>0.997) over a concentration range of 2.5-200 ${\mu}g/ml$. The peak of the excreted cetirizine enantiomers appeared in the urine sample during the period of 1-2 hrs following the administration of the oral dose. The excreted level of (+)-cetirizine was slightly higher than (-)-cetirizine but the difference was not statistically significant. However, this method appears to have applications for enantioselective pharmacokinetic studies of racemic drugs.

• Journal of Microbiology and Biotechnology
• /
• v.23 no.4
• /
• pp.451-458
• /
• 2013

The human gastric pathogen Helicobacter pylori (Hp) has been considered a microaerophile. However, we recently reported that, when supplied with 10% $CO_2$, Hp growth is stimulated by an atmospheric level of $O_2$, suggesting that Hp is a capnophilic aerobe. In this study, we investigated the effects of aerobic $O_2$ tension on Hp cells by comparing gene expression profiles of cultures grown under microaerobic and aerobic conditions in the presence of 10% $CO_2$. The results showed that overall differences in gene expression in Hp cells grown under the two $O_2$ conditions were predominantly growth-phase-dependent. At 6 h, numerous genes were down-regulated under the aerobic condition, accounting for our previous observation that Hp growth was retarded under this condition. At 36 h, however, diverse groups of genes involved in energy metabolism, cellular processes, transport, and cell envelope synthesis were highly up- or down-regulated under the aerobic condition, indicating a progression of the cultures from the log phase to the stationary phase. The expression of several oxidative stress-associated genes including tagD, katA, and rocF was induced in response to aerobic $O_2$ level, whereas trxA, trxB, and ahpC remained unchanged. Altogether, these data demonstrate that aerobic $O_2$ tension is not detrimental to Hp cells but stimulates Hp growth, supporting our previous finding that Hp may be an aerobic bacterium that requires a high $CO_2$ level for its growth.

• Molecules and Cells
• /
• v.22 no.3
• /
• pp.336-342
• /
• 2006

Real time reverse transcription (RT)-PCR was used to quantify the expression of the botulinum neurotoxin type A (BoNT/A) gene (cntA) by normalization with the expression of 16S rRNA. The method were confirmed by monitoring the mRNA levels of cntA during growth in five type A strains. In all but one of the strains the expression of cntA mRNA was maximal in the late exponential phase, and approximately 35-fold greater than in the early exponential phase. The concentration of the extracellular BoNT/A complex detected by ELISA was highest in stationary phase. Sodium nitrite and sorbic acid completely inhibited growth at 20 ppm and $4mg\;ml^{-1}$, respectively. CntA expression became lower in proportion to the concentration of sorbic acid, and this reduction was confirmed by mouse bioassay. Our results show that real time RT-PCR can be used to quantify levels of C. botulinum type A neurotoxin transcripts and to assess the effects of food additives on botulinal risk.

• Korean Journal of Microbiology
• /
• v.19 no.2
• /
• pp.63-77
• /
• 1981

The growth rate of yeast population (Saccharomyces uvarum) cultivated in the Knopp's modified medium (plus various carbon sources) appeared the highest value when the Knopp's minimal medium was treated to 1.5% with disaccharide such as maltose and sucrose. Also the treatment of lactose and raffinose resulted in polulation growth as to the population size in case of maltose and sucrose. However, the gorwth of yeast was not occurred at all when a polysaccharide, such as inulin, was added as carbon source. The growth from of yeast population in Knopp's modified medium are characterized by the fact that log phase continued 100hrs after inoculation and that stationary state phase appeared in general 250hrs after inoculation. Applying the various carbon sources to respiration substrate for yeast cell, the respiration rate of yeast showed the highest value in treatment of maltose and followed in order of raffinose, lactose, glucose, and sucrose. Determined the amount of poly-phosphate and turn over pathway of poly-phosphate according to culture phase of yeast, it is revealed that the yeast synthesized 3 types of poly phosphate (poly-P A,B, and C) and postulated that turn over pathway of poly-phosphate as follows ; Inorganic phosphate is converted into each kind of polyphosphates, and then one part of poly-P-C is converted into poly-P-B, the rest poly-p-C and poly-P-B are converted into poly-P-A. The synthesized poly-phosphate is considered to have a role as energy pool utilizing to synthesis of cellular organic materials. Of the 13 carbon sources used in this experiment, the useful carbon sources for biosynthesis of poly-phosphate and cellular organic materials are confirmed as disaccharide (maltose and sucrose) as well as glucose. Protein synthesis in yeast cell showed the two peaks on 6th and 8th day after inoculation ; nucleic acid on 2nd day (48hrs), carbohydrates on 2nd day (48hrs), and phospholipid on 2nd and 8th day after inoculation, respectively.

• Korean Journal of Optics and Photonics
• /
• v.14 no.6
• /
• pp.606-612
• /
• 2003

The self-imaging effect or lensless imaging effect of a phase line grating is theoretically analyzed by using Fresnel diffraction theory, then experimentally investigated. The self-imaging distance $z_{T,p}$, that is the imaging distance being perfectly copied from the phase distribution of the phase grating to its intensity distribution with the magnification of 1X, can be uniquely defined as the (4n-3) $z_{T,a}$/4(n=positive integers), where rte is the well-known self-imaging distance of an amplitude grating. When the coherent laser beam is illuminated at the phase grating, the self-imaged images were obtained at $z_{T,p}$= $z_{T,a}$/4 and $z_{T,p}$=5 $z_{T,a}$/4 without any optics. On the other side, the phase-reversed self-imaging was obviously observed at $z_{T,p}$ = 3 $z_{T,a}$/4. The visibility of self-imaged images of a phase line grating as a function of the number of slits of the input grating was measured by the FFT(Fast Fourier Transform) results of the self-imaging images. As a result a stationary maximum visibility of V = 0.10 can be obtained from a grating with more than 15 slit pairs.n 15 slit pairs.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.13 no.11
• /
• pp.5179-5186
• /
• 2012

Miglitol, a well-known therapeutic intervention agents for diabetes, exhibits competitive inhibitory activity against ${\alpha}$-glucosidase and it is usually produced through three sequential steps including chemical and bioconversion processes. Gluconobactor oxydans (G. oxydans) belonging to acetic acid bacteria biologically, converts 1-deoxy-1-(2-hydroxyethylamino)-D-glucitol (P1) into a key intermidiate, 6-(2-hydroxyetyl) amino-6-deoxy-${\alpha}$-L-sorbofuranose (P2) by incomplete oxidation. In this study, we identified and optimized fermentation conditions of CK-2165, that was selected in soil samples by comparing the bioconversion yield. CK-2165 strain was found to be closely related to G. oxydans based on the result of phylogenetic analysis using 16S rDNA sequence. Utilization of API 20 kits revealed that this strain could use glucose, mannose, inositol, sorbitol, rhamnose, sucrose, melibiose, amygdalin and arabinose as carbon sources. The culture conditions were optimized for industrial production and several important factors affecting bioconversion rate were also tested using mycelial cake. Cell harvested at the late-stationary phase showed the highest bioconversion yield and $MgSO_4$ was critically required for the catalytic activity.

• Journal of Food Hygiene and Safety
• /
• v.19 no.3
• /
• pp.151-160
• /
• 2004

Enterococcus faecium MJ5-14 isolated from Meju produced a bacteriocin, which was antagonistic towards Listeria monocytogenes. Bacteriocin activity reached a maximum (640 BU/mL) after incubation for 12 hr, the early stationary phase, then dropped after the late stationary phase. Bacterocin of E. faecium MJ5-14 was extremely active against a wide range of Listeria species, including L. monocytogenes with sensitives up to about 640 BU/mL. In case of mixed culture with 105 CFU/mL L. monocytogenes and 105 CFU/mL E. faecium MJ5-14, the inhibitory effect against L. monocytogenes at $37^{\circ}C$ was higher than at $25^{\circ}C$. The mode of action was identified as bactericidal, because the addition of 100 BU/mL this bacteriocin to cell suspensions of L. monocytogenes KCTC 3569, led to a marked decrease in the number of viable cells. Further, when held in contact with bacteriocin of E. faecium MJ15-14 for 12 hr, L. monocytogenes KCTC 3569 displayed the disruption of the cells and an important efflux of the intracellular material.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.12
• /
• pp.1345-1351
• /
• 2011

Vancomycin therapy failure due to the emergence of tolerance in pneumococci is increasing. The molecular mechanism of tolerance is not clear, but lytA and $pep^{27}$ are known to be involved. Our aim was to evaluate the expression of both genes in vancomycin-tolerant Streptococcus pneumoniae (VTSP) strains. Eleven VTSP strains from a total of 309 clinical isolates of S. pneumoniae from 1997 to 2006 were classified according to the criteria of Liu and Tomasz. All VTSP strains were evaluated for susceptibility according to CLSI criteria, serotype by the Quellung test, and clonality by PFGE. The expressions of lytA and $pep^{27}$ were analyzed in different growth phases by RT-PCR with and without vancomycin. Eighty-two percent of VTSP strains showed resistance to penicillin, and 100% were sensitive to vancomycin and cefotaxime. The most frequent serotypes of VTSP strains were 23F (4/11) and 6B (3/11). Clonal relationship was observed in only two strains. No significant changes were observed in $pep^{27}$ expression in the three phases of growth in VTSP strains with and without vancomycin. Interestingly, $pep^{27}$ expression in the stationary phase in the non-tolerant reference strain R6 was significantly higher. However, no significant differences in lytA expression were observed between VTSP and R6 strains during the phases of growth analyzed. The absence of changes in $pep^{27}$ expression in VTSP strains in the stationary phase may be related to their ability to tolerate high antibiotic concentrations, and thus, they survive and remain in the host under the antibiotic selective pressure reflected in therapeutic failure.