• Biomedical Science Letters
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• v.9 no.3
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• pp.139-144
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• 2003

Since water borne infection causes acute diseases and results in spread of diseases by secondary infection, the prevention is very important. Therefore, it is necessary to have a method that is rapid and effective to monitor pathogenic bacteria in drinking water. In this study, we employed a systematic method, Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis, to develop an effective monitoring system for possible bacterial contaminants in drinking water. For this purpose, PCR primers were derived from 992 bp region of the 16s rRNA gene that is highly conserved through the different species of prokaryotes. To test whether the PCR primers designed are indeed useful for detecting all the possible microbial contaminants in the water, the primers were used to amplify 16s rRNA regions of different microbial water-borne pathogens such as E. coli, Salmonella, Yersinia, Listeria, and Staphylococcus. As expected, all of tested microorganisms amplified expected size of PCR products indicating designed PCR primers for 16s rRNA indeed can be useful to amplify all different microbial water-borne pathogens in the water. Furthermore, to test whether these 16s rRNA based PCR primers can detect bacterial populations present in the water, water samples taken from diverse sources, such as river, tap, and sewage, were used for amplification. PCR products were for then subjected for cloning into a T-vector to generate a library containing 16s rRNA sequences from various bacteria. With cloned PCR products, RFLP analysis was done using PCR products digested with restriction enzyme such as Hae III to obtain species-specific RFLP profiles. After PCR-RFLP, the bacterial clones which showed the same RFLP profiles were regarded as the same ones, and the clones which showed distinctive RFLP profiles were subsequently subjected for sequence analysis for species identification. By this PCR-RFLP analysis, we were able to reveal diverse populations of bacteria living in water. In brief, in unsterilized natural river water, over 60 different species of bacteria were found. On the other hand, no PCR products were detected in drinking tap-water. The results from this study clearly indicate that the PCR-RFLP-sequence analysis can be a useful method for monitoring diverse, perhaps pathogenic bacteria contaminated in water in a rapid fashion.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.293-297
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• 2012

Of the 169 strains of microorganisms stored in Polar and Alpine Microbial Collection of Korea Polar Research Institute, 27 strains were selected for their chitinase activity on ZoBell plates supplemented with 0.4% colloidal chitin. Among them, PAMC 21693 strain have shown the highest chitinolytic enzyme activity toward pNP-$(GlcNAc)_1$ at low temperature and the highest growth rate at $4^{\circ}C$. We cloned a full-length chitinase gene of 2,857 bp which contains an open reading frame of 2,169 bp encoding 872-amino acid polypeptide. Recombinant chitinase protein was expressed in E. coli and its molecular weight was confirmed 96 kDa. In this paper, we suggest the potential use of cold-active chitinase from polar microorganisms in the field of biotechnology.

• KSBB Journal
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• v.24 no.5
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• pp.432-438
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• 2009

Here, we demonstrated simple nucleic acid, RNA, concentration method using polymer micro chip containing glass bead ($100\;{\mu}m$). Polymer micro chip was fabricated by PDMS ($1.5\;cm\;{\times}\;1.5\;cm$, $100\;{\mu}m$ in the height) including pillar structure ($160\;{\mu}m\;(I)\;{\times}\;80\;{\mu}m\;(w)\;{\times}\;100\;{\mu}m\;(h)$, gap size $50\;{\mu}m$) for blocking micro bead. RNA could be adsorbed on micro glass bead at low pH by hydrogen bonding whereas RNA was released at high pH by electrostatic force between silica surface and RNA. Amount of glass beads and flow rate were optimized in aspects of adsorption and desorption of RNA. Adsorption and desorption rate was measured with real time PCR. This concentrated RNA was applied to amplification micro chip in which NASBA (Nucleic Acid Sequence Based Amplification) was performed. As a result, E.coli O157 : H7 in the concentration of 10 c.f.u./10 mL was successfully detected by these serial processes (concentration and amplification) with polymer micro chips. It implies this simple concentration method using polymer micro chip can be directly applied to ultra sensitive method to measure viable bacteria and virus in clinical samples as well as environmental samples.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.19 no.9
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• pp.384-390
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• 2018

Air and soil are being contaminated by the environmental pollution as a result of climate change and urbanization, resulting in water pollution reaching serious levels. In this studies, we investigated the use of antimicrobial copper for the removal of biological pollutants from water system. Specifically, we tested its effects against E. coli, B. subtilis, S. aureus, K. pneumoniae and P. aeruginosa. Made a sphere shape having a diameter of 2cm using a strip-shaped copper wire of 0.5g, 1g and 2g. And then, to confirm the antimicrobial activities, each copper ball was equipped in the broth which inoculated each pathogens. The results showed that bacterial growth of the five test bacteria was inhibited by more than 99% after reaction with a 0.5 g copper ball for at least 20 minutes. Based on the these results, if perform the further experiment such cytotoxicity, it is expected that will be enough to be used as a filter for water quality purification. The developed technique is expected to be widely applied in various industries.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.7
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• pp.1053-1058
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• 2003

The solvent extracts of Houttuynia cordata root, which were extracted by using several solvents with different polarities, were prepared for utility as a natural preservatives. The antimicrobial activity was investigated by disc diffusion method against 22 microorganisms consisting of food borne pathogens, food poisioning microorganisms and food-related bacteria. The extraction yields were 15.7%, 3.7%, 0.13%, 0.5% and 5.9% in ethanol, chloroform ethylacetate, butanol and aqueous fractions, respectively. Antimicrobial activities were shown in ethanol, ethylacetate and butanol fraction of Houttuynia cordata root. However chloroform and aqueous fractions showed weak antimicrobial activity against the tested bacteria. Among the four fractions, ethylacetate fraction showed the strongest antimicrobial activities against microorganisms tested, such as B. megaterium, E. coli, K. pneumoniae and S. typhimurium. The polyphenolic compounds widely occuring in the traditional medicine plants have been reported to possess high antimicrobial activity. The polyphenolic compound in ethylacetate and butanol fraction were 35.9% and 16.0%, ethanol, chloroform and aqueous fraction were 5.0%, 2.3% and 1.7%, respectively. There are some relationship between antimicrobial activity and polyphenol content in natural plants. The ethylacetate fraction could be suitable for the development of a food preservative.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.3
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• pp.394-400
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• 2004

To investigate the efficacy of alternatives to antibiotics, the present study was conducted to compare the effects of antibiotic, lactic acid, a blend of commercial essential oils (EOs) and EOs in combination with lactic acid on growth performance and the functional activity of the gut in broiler chickens. A total of 168 broiler chickens were given the basal diet supplemented with 10 ppm colistin (T1), 0.1% lactic acid (T2), 25 ppm EOs (T3), 25 ppm EOs+0.1% lactic acid (T4), 50 ppm EOs (T5) or 50 ppm EOs+0.1% lactic acid (T6) in the period 3 to 35 days of age. As a result, the broiler chickens assigned to T4 group throughout the experimental period had apparently (p<0.05) greater body weight and total gain than these assigned to T1, T2, T3 and T5 groups. However, there was no difference in growth performance among the birds fed the diets supplemented with antibiotic (T1), lactic acid (T2) and EOs (T3 and T5) alone. The weights of digestive organs and the number of lactobacilli and E. coli in the lower ileum were not affected by dietary treatments. Total trypsin activity was significantly (p<0.05) greater in T4 than T1, T2, T3 and T5 groups. Total and specific pancreatic $\alpha$-amylase activities were significantly (p<0.05) enhanced in the broiler chickens fed T4 diet compared with these fed T1, T2 and T3 diets. However, there were no differences in growth performance and digestive enzyme activities including pancreatic trypsin and $\alpha$-amylase between T4 and T6 groups fed the diets supplemented with either low or high EOs levels in combination of lactic acid. In conclusion, a blend of commercial EOs combined with lactic acid showed significant increases in digestive enzyme activities of the pancreas and intestinal mucosa, leading to increase in growth performance.

• Journal of Microbiology and Biotechnology
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• v.23 no.6
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• pp.818-825
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• 2013

We isolated and functionally characterized the ${\alpha}$- and ${\beta}$-subunits (ApCpnA and ApCpnB) of a chaperonin from Aeropyrum pernix K1. The constructed vectors pET3d-ApCpnA and pET21a-ApCpnB were transformed into E. coli Rosetta (DE3), BL21 (DE3), or CodonPlus (DE3) cells. The expression of ApCpnA (60.7 kDa) and ApCpnB (61.2 kDa) was confirmed by SDS-PAGE analysis. Recombinant ApCpnA and ApCpnB were purified by heat-shock treatment and anion-exchange chromatography. ApCpnA and ApCpnB were able to hydrolyze not only ATP, but also CTP, GTP, and UTP, albeit with different efficacies. Purified ApCpnA and ApCpnB showed the highest ATPase, CTPase, UTPase, and GTPase activities at $80^{\circ}C$. Furthermore, the addition of ApCpnA and ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively. In particular, the addition of ATP or CTP to ApCpnA and ApCpnB resulted in the most effective prevention of thermal aggregation and inactivation of CS and ADH. The ATPase activity of the two chaperonin subunits was dependent on the salt concentration. Among the ions we examined, potassium ions were the most effective at enhancing the ATP hydrolysis activity of ApCpnA and ApCpnB.

• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1749-1759
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• 2018

Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

• Journal of Animal Science and Technology
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• v.59 no.8
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• pp.18.1-18.8
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• 2017

Background: In 2006, the European Union (EU) has decided to forbid use of antibiotics as growth promoters. Although many researches had been conducted about fiber source as alternatives of antibiotics, there are still lack of reports in the literature about the optimum level of sugar beet pulp supplementation, affecting growth performance and nutrient digestibility in weaning pigs. Therefore, different level of sugar beet pulp was added to diets to determine the effects of sugar beet pulp supplementation on growth performance, nutrient digestibility, fecal microflora, blood profile and incidence of diarrhea in weaning pigs. Methods: A total of 200 weaning pigs [$(Yorkshire{\times}Landrace){\times}Duroc$], averaging $9.01{\pm}1.389kg$ of initial body weight were, allotted to 5 treatments in a randomized complete block (RCB) design. Each treatment was composed of 4 replicates with 10 pigs per pen. The treatments were control treatment: Corn-SBM basal diet + ZnO (phase 1: 0.05%; phase 2; 0.03%) and four different levels of sugar beet pulp were supplemented in Corn-SBM basal diet (3, 6, 9 or 12%). Two phase feeding programs (phase 1: 1-2 weeks; phase 2: 3-5 weeks) were used for 5 week of growth trial. Results: In feeding trial, there were no significant differences in growth performance and incidence of diarrhea among treatments. The E.coli counts were not significantly different among dietary treatments but linear response was observed in Lactobacillus counts as sugar beet pulp supplementation increased (P < 0.05). In addition, IGF-1, IgA and IgG were not affected by dietary treatments. However, the BUN concentration was decreased when pigs were fed the treatments of diets with SBP compared to that of control treatment (P < 0.05). In nutrient digestibility, crude fiber and NDF digestibilities were improved as the sugar beet pulp increased (P < 0.05). However, digestibilities of crude ash, crude fat, crude fiber and nitrogen retention were not affected by dietary sugar beet pulp levels. Conclusion: This experiment demonstrated that sugar beet pulp can be supplemented in weaning pigs' diet instead of ZnO to prevent postweaning diarrhea without any detrimental effect on growth performance.

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.196-203
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• 2006

Three slime-forming lactic acid bacteria were isolated from Kimchi and shown to produce viscous exopolysaccharides (EPS) in sucrose media. The isolated strains, GJ2, C3 and C11, were identified as Leuconostoc kimchii, Leuconostoc citreum and Leuconostoc mesenteroides, respectively, by examining their metabolic characteristics and determining their 16S rDNA sequences. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 exhibited high viability (maintained initial viable cell count of $10^8$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastric juice for 2 h and in 0.3% oxgall for 24 h. When tested, Leu. kimchii GJ2, in particular, displayed antimicrobial activity against pathogenic microorganisms. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 produced 21.49 g/l, 16.46 g/l and 22.98 g/l EPS, respectively, in sucrose (5%) medium. The amount of purified EPS extracted from Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 was 14.61 g/l, 7.73 g/l and 4.77 g/l, respectively. Although the EPS produced by Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 differed in viscosity, TLC and HPLC analysis revealed that each contained only one type of monosaccharide, glucose. The average molecular mass of EPS produced by Leu. kimchii GJ2 was 306,606 Da.