• KSBB Journal
• /
• v.20 no.5 s.94
• /
• pp.376-382
• /
• 2005

Avermectin (AVM) $B_{1a}$ produced by Streptomyces avermitilis via polyketide pathway is a secondary metabolite with powerful anthelmintic and insecticidal activities, thus being used as an efficient agent in the field of agriculture and animal health. It has been reported that a precursor for AVM $B_{1a}$ biosynthesis was isoleucine and the biosynthetic pathway of AVM $B_{1a}$ was closely similar to that of fatty acid. Based on understanding of the biosynthetic pathway of AVM $B_{1a}$, we intended to screen various mutants resistant against O-methyl threonine (OMT), an isoleucine-anti metabolite, and/or mutants resistant against p-fluoro phenoxy acetic acid (pFAC), an inhibitor of fatty acid biosynthesis. It was inferred that these mutants could produce AVM $B_{1a}$ more efficiently, due to the acquired capability of not only overproducing isoleucine intracellularly but also channelling metabolized carbon-sources into the polyketide pathway, thus leading to enhanced biosynthesis of AVM $B_{1a}$. The resulting mutant (PFA-1 strain) resistant against 100 ppm of pFAC was able to produce approximately 42 fold higher amount of AVM $B_{1a}$ compared to the parallel mother strain (4,200 vs. 100 units/l). In addition, through the process of continuous strain improvement program carried out by gradually increasing the OMT concentration, it was possible to obtain a more attractive mutant with greater AVM $B_{1a}$ production capacity (9,000 units/l). Notable was that significantly higher producer (12,000 units/l) could be selected through further screening of the resistant mutants, this time, to even higher concentration of PFAC. Meanwhile, through the analysis of AVM Bla production histograms (i.e., number of strains according to their AVM $B_{1a}$ biosynthetic ability) for the earlier strains in comparison with the high producers having the characteristics of resistance to OMT and pFAC, it was found that production stability of the high-yielding producers were remarkably improved, as demonstrated by the fact that larger proportion of the mutated strains had greater capability of AVM $B_{1a}$ biosynthesis ($71\%$ in the range between 5,000 and 7,000 units/L; $47\%$ in the range between 6,000 and 7,000 units/l). Based on these consequences, it was concluded that the rational screening strategy based on the understanding of the biosynthetic pathway of AVM $B_{1a}$ was very effective in obtaining high-yielding mutants with the features of enhanced production stability.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
• /
• 2004.05a
• /
• pp.89-119
• /
• 2004

This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Journal of Acupuncture Research
• /
• v.22 no.3
• /
• pp.37-51
• /
• 2005

Mast cell is a cell that functions mainly in our body with a respect to inflammation and allergic response. Bee venom has been progressed in a study as a model related to mechanism in alleviation of pain until now, but it is being progressed in a study relevant to immunocyte in anti-inflammation or anti-allergic response. The present study is aimed to present the basis related to a future study of gene, by researching the influence of melittin and MCD Peptide, which are major ingredients in Bee venom, upon the expression of gene in the mast cell strain. In this study, it dealt with melittin and MCD Peptide respectively, in the effective concentration after passing though the experiment of cytotoxicity by using human mast cell strain. Also, with the respect in the aspect of expression in gene that changes at this time, information was obtained through the technique of analyzing microarray. Through experimental statistics, when regarding a case that global M is significant in more than 1 or -1, in melittin, all 7 genes were accelerated, and 8 inhibited. In MCDP, 7 genes were accelerated and 17 genes inhibited. The function in the body to which these genes are related, was associated with the protein binding within a cell, the activation in the function of lymphocyte, the acceptor related to macrophage antigen. In cell nucleus, substance related to GABA A receptor, protein associated with cAMP reactive element, substance related to complement system No.8 and to B-cell, protein substance related to polycystic kidney disease, substance related to inflammation, and the protein substance of influencing coagulation of blood. Through these results of analysis, it could obtain more useful materials in clarifying the mechanism of action in melittin and MCD peptide, which are in charge of mainly medical action in the abdomen. Also, it is thought that an in-depth study on the influence of main ingredients in Bee venom, the wholly honey bee venom aqua upon anti-allergic response or anti-inflammation are further required.

• Food Science and Preservation
• /
• v.23 no.7
• /
• pp.1050-1057
• /
• 2016

This study was carried out to investigate the effectiveness of Leuconostoc mesenteroides isolated from octopus baechu kimchi as a potential starter for seafood kimchi. L. mesenteroides is lactic acid bacterium currently used as a starter for kimchi production. We selected the most effective L. mesenteroides strain from the 7 strains isolated from octopus baechu kimchi and, based on biochemical properties and 16S rRNA sequencing, identified the selected strain as L. mesenteroides SK-1. The SK-1 strain exhibited acid-tolerance, good survival capacity, and excellent dextran productivity. We investigated the effects the SK-1 of starter on seafood kimchi fermentation. Octopus baechu kimchi was fermented with L. mesenteroides SK-1 at $4^{\circ}C$ for 35 d. The decrease in pH and increase in acidity in octopus baechu kimchi fermented with the SK-1 starter occurred more quickly than that in the control kimchi indicating that. Octopus baechu kimchi with SK-1 starter has a relatively slow rate of increase in lactic acid production. As a result, octopus baechu kimchi prepared with L. mesenteroides SK-1 can be maintained at a suitable ripening degree over an extended period of time compared to that of the control kimchi, Moreover, the octopus baechu kimchi started with L. mesenteroides SK-1 has excellent sensory properties, including a refreshing taste, and a weak sour odor.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.3
• /
• pp.591-599
• /
• 2008

A murine model immunized by systemic and mucosal delivery of plasmid DNA vaccine expressing glycoprotein B (pCIgB) of pseudorabies virus (PrV) was used to evaluate both the nature of the induced immunity and protection against a virulent virus. With regard to systemic delivery, the intramuscular (i.m.) immunization with pCIgB induced strong PrV-specific IgG responses in serum but was inefficient in generating a mucosal IgA response. Mucosal delivery through intranasal (i.n.) immunization of pCIgB induced both systemic and mucosal immunity at the distal mucosal site. However, the levels of systemic immunity induced by i.n. immunization were less than those induced by i.m. immunization. Moreover, i.n. genetic transfer of pCIgB appeared to induce Th2-biased immunity compared with systemic delivery, as judged by the ratio of PrV-specific IgG isotypes and Th1- and Th2-type cytokines produced by stimulated T cells. Moreover, the immunity induced by i.n. immunization did not provide effective protection against i.n. challenge of a virulent PrV strain, whereas i.m. immunization produced resistance to viral infection. Therefore, although i.n. immunization was a useful route for inducing mucosal immunity at the virus entry site, i.n. immunization did not provide effective protection against the lethal infection of PrV.

• Journal of Korean Society of Steel Construction
• /
• v.22 no.5
• /
• pp.399-409
• /
• 2010

Steel structures are threatened to reduce load-carrying capacity as the cross section is decreased by corrosion. However, there has been no method in definitely evaluating residual load-carrying capacity and the effect of corrosion to the load-carrying capacity of steel. This study evaluated tensile residual load-carrying capacity of corroded steel plates by using tensile tests of specimens, which were selected from the web of temporary structure's main beam. After the surface shapes were measured and tensile tests were examined, the rust of 21 corroded specimens was, first of all, removed using a chemical method. From the tensile test result, which of reference specimens that was picked off at the flange of the same main 13-mm-thick beam and corroded specimens were based, surface geometry and correlation with the reduction of corroded thickness and strain, yield strength or tensile strength was established as constant numbers. Effective thickness of corroded steel with irregular cross sections could be calculated using average residual thickness and standard deviation. The irregular cross sections could be the evaluated tensile strength that is equalized to non-corroded uniform steel's regardless of corrosion. Also, reasonable measuring intervals of residual thickness could be proposed by using this result to apply for executive work.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.37 no.4
• /
• pp.347-350
• /
• 2011

Biofilms are surface-attached microbial communities with phenotypic and biochemical properties distinct from free-living planktonic cells. Biofilm bacteria show much greater resistance than planktonic counterparts and much higher concentration of biocide is needed to treat biofilms compared to the dosage used for planktonic bacteria. As a result, alternative strategies or more effective agents exhibiting activity against biofilm-producing micro-organisms are of great interest. Therefore, we turned our attention to control of biofilm of S. aureus. The aims of this research are to investigate substances which inhibit the formation of biofilm by S. aureus and to suggest effective materials for controlling skin problems. We coated slide glasses with human placental collagen and the coverslip was incubated with test materials and bacteria. The coverslip was stained with crystal violet and we measured optical density of each sample. The biofilm inhibitory activity was calculated by crystal violet staining degrees. In this study, S. aureus ATCC 6538 was used as test organism. Our results show that both water soluble and insoluble Hinoki cypress polysaccharide strongly inhibited biofilm formation. Whereas, green tea and sunset hibiscus root extract promoted biofilm. Xylitol showed a concentration dependent effect; high concentration (3 % and 5 %) of xylitol reduced biofilm while promoted biofilm formation at a concentration of 1 %. These results support that Hinoki cypress polysaccharide and xylitol have ability to suppress biofilm formation.

• Journal of Life Science
• /
• v.15 no.5 s.72
• /
• pp.755-762
• /
• 2005

Biochemical amounts of RNA molecules can be synthesized in vitro, which is functionally equivalent or similar to those transcripts normally existing at extremely low levels in vivo. In this study we described a method for efficient preparation of pure T7 RNA polymerase from Escherichia coli strain BL21/pAR1219. The procedure, which used ammonium sulfate fractionation and preparative column chromatography on sephadex SP, was shown to be simple, rapid, and cost effective in comparison with other methods reported previously, Using the purified T7 RNA polymerase we were able to synthesize very long RNA transcript of 1.54 kb length, which is not feasible by conventional chemical synthesis. RNA molecule that was also synthesized by the purified T7 RNA polymerase, such as hammerhead ribozyme, retained its biochemical activity by cleaving the target RNA successfully in vitro. Thus, the procedure shown in this study can be useful to synthesize any length of RNA molecules in vitro in a simple and cost effective way for a variety of purposes.

• The Korean Journal of Pesticide Science
• /
• v.18 no.4
• /
• pp.358-363
• /
• 2014

We was investigated parasporal inclusion proteins change to use industrial medium of new strain Bacillus thuringiensis CAB 565, CAB 566. To confirm medium's oxygen efficient consist of glucose and yeast extract, we was conducted oxygen transfer coefficients (KLa) of medium's concentration and impeller in 20 l-Jar fermentor. When to increase air flow rate and medium concentration, KLa rate is rise. Also it is effective on agitation rate 200 rpm, but KLa rate is decrease when to rise agitation rate. To hold dissolved oxygen rate (upper 50%), Air flow rate is steadily increase on culture to use microsparger. When 16 hour of culture stage, B.t. CAB 565 and B.t. CAB 566 harvested respectively $2.3{\times}10^{10}$, $1.8{\times}10^{10}$ viable cell/ml. When 54 hour, B.t. CAB565, 566 harvested respectively $1.9{\times}10^{10}$, $1.4{\times}10^{10}spore/ml$. To resulting carbon's concentration, It is the most effective that glucose concentration is contained 5% in medium.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.12 no.3
• /
• pp.161-166
• /
• 1979

To know the influence of temperature on the fermentation process, a strain of Lactobacillus bulgarius was experimentally cultured three different temperature conditions of $39^{\circ}C,\;42^{\circ}C\;and\;45^{\circ}C$, pH 5.8 and mechanical agitation of 500rpm. During 20 hour's fermentation, the microbial growth attained the maximum concentration under the conditions mentioned above. However, the culturing conditions resulted different outcomes in terms of maximum concentration of the microbes and the residual concentration of substrate. Among the three temperature conditions, the fermentation at $45^{\circ}C$ was most effective and the maximum specific growth temperature conditions, the fermentation at $45^{\circ}C$ was most effective and the maximum specific growth rate was 0.58/hr. Activation energy deduced from the Arrhenius equation was 9,220cal/mole and entropy was $-33.74\;cal/^{\circ}K$ mole. Activation enthalpy was 9,845 cal/mole and free energy was 19,800 cal/mole.