• Korean Journal of Environmental Agriculture
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• v.12 no.1
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• pp.41-49
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• 1993

The Study was conducted to develope the low temperature tolerant methane-producing bacteria(LTTB) and to increase the efficiency of anaerobic fermentation for the treatment of agricultural and livestock wastes at low temperature. The samples were collected from muddy soil, water logged sediment, organic layer and anaerobic sludge at three latitudes, $34.8{\sim}37.4\;^{\circ}N(Korea)$, $41.4\;^{\circ}N(USA)$ and $54.5{\sim}56.9\;^{\circ}N(Canada)$. They were used for determination of the methanogenesis rates for isolation and identification of the LTTB. The methanogenesis rate of smaples at low temperature were higher in the cellulose medium than methanol medium. The methanogenesis rate in the samples of subarctic region were $15{\sim}19$ moles/ml during 30 days at low temperature($8\;^{\circ}C$), whereas not detected in the samples of temperate region. The methanogenesis rate in the enrichment culture of subarctic samples were inhibited by the $40\;{\mu}g/ml$ of streptomycin + vancomycin or ampicillin + oleandomycin which were not effect to the methanogens. An inhabitation of high temperature tolerant methane producing bacteria was identified in the samples of temperate region, whereas that of the LTTB growing at $8{\sim}13^{\circ}C$ was identified in the subarctic region.

• Korean Journal of Environmental Agriculture
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• v.15 no.3
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• pp.362-371
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• 1996

This study was conducted to investigate the biochemical properties of isolated bacteria, low temperature tolerant methanogens which were selected for use as inoculum for anaerobic fermentation of agricultural and livestock wasted at low temperature. The results, obtained were summarized as follows: Low temperature tolerant methanogens were isolated from the samples which showed the high methanogenesis rate by enrichment culture at low temperature in methanol medium. These methanogens, Methanobacterium M-251 and Methanobacterium M-253 were isolated from swampy sediment at latitude $56.9^{\circ}$, Methanosarcina mazei M-372 from lake sediment IV at latitude $55.0^{\circ}$ N, and Methanobacterium formicicum M-375 from tidal land soil at latitude $37.0^{\circ}N$, respectively. The isolated anaerobic bacteria could not use sugars as carbon sources. The optimum pH value for the growth of M-251 and M-375 was 6.8, but those for M-253 and M-372 6.5 and 7.0, respectively. The minimum growth temperature of isolated, M-251 and M-253 were $8^{\circ}C$ and the optimum temperature $30^{\circ}C$, while the minimum of M-392 and M-395 were $13^{\circ}C$ and the optimum $37^{\circ}C$. The growth rate of isolates at $17.5^{\circ}C$ were lower by 32-50% than that of $30^{\circ}C$. The isolated Methanobacterium strains such as M-251, M-253, and M-375 have lower cell yield, 0.38-1.21g/1M $CH_4$ than 1.14-1.51g/1M $CH_4$ of Methanosarcina mazei M-372.

• Restorative Dentistry and Endodontics
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• v.25 no.4
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• pp.561-574
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• 2000

Poly-P has been used to prevent decomposition of foods and has been shown to have inhibitory effect on the growth of gram positive bacteria. The purpose of this study was to evaluate the effect of poly-P on the growth of Porphyromonas endodontalis, a gram negative obligate anaerobic rod, endodontopathic bacterium. P. endodontalis ATCC 35406 was in BHI broth containing hemin and vitamin K with or without poly-P. Inhibitory effect of each poly-P which was added at the beginning(lag phase) or during(exponential phase) the culture, MIC(minimum inhibitory concentration) was determined by measuring the optical density of the bacterial cell at 540nm. Viable cell counts were measured to determined whether poly-P has a bactericidal effect. Leakage of intracellular nucleotides from P. endodontalis was determined at 260nm and morphological change of P. endodontalis was observed under the TEM(transmission electron microscope). Binding of 32P-labeled poly-P to P. endodontalis was examined. SDS-polyacrylamide gel electrophoresis and zymography were performed to observe the changes in protein and enzyme profiles of P. endodontalis, respectively. The results from this study were as follows : 1. The minimal inhibitory concentration(MIC) of poly-P to P. endodontalis appeared to be 0.04~0.05%. 2. Poly-P added to the P. endodontalis culture during the exponential phase of P. endodontalis was as much effective as poly-P added at the begining of the culture, suggesting that the antibacterial effect of poly-P is not much dependent on the initial inoculum size of P. endodontalis. 3. Poly-P are bactericidal to P. endodontalis, demonstrating the decrease of the viable cell counts. 4. Intracellular nucleotide release from the P. endodontalis, was not increased in the presence of poly-P and was not reversed by the addition of divalent cations like $Ca^{2+}$ and $Mg^{2-}$. 5. Under the TEM, it was observed that fine electro-dense materials were prominent in the poly-P grown P. endodontalis, appearing locally in the cell, and the materials were more abundant and more dispersed in the cell as the incubation time with poly-P increased. In addition, highly electron dense granules accumulated in many poly-P grown cells, most of which were atypical in their shape. 6. Binding of 32P-labeled poly-P to P. endodontalis appeared to be 32.8 and 45.5 and 53.4% at 30 minutes, 1 hours and 2 hours, respectively. 7. In the presence of poly-P. the synthesis of proteins with apparent molecular masses of 25, 27, 35, 45 was lost or drastically decreased whereas expression of a protein with an apparent molecular mass of 75 was elevated. 8. Proteolytic activity of P. endodontalis was decreased by poly-P. The overall results suggest that use of poly-P may affect the growth of P. endodontalis, and the anti-bacterial activity of poly-P seems largely bactericidal. Changes in shape, protein expression, and proteolytic activity of P. endodontalis by poly-P may be directly and indirectly attributed to the antibacterial effect of poly-P. Further studies will be needed to confirm the effect of poly-P.

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.2
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• pp.344-356
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• 2005

There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic acid bacteria were isolated from inhabitants of caries-free children's oral cavity, which inhibited the formation of artificial plaque by Streptococcus mutans and the production of volatile sulfur compounds by anaerobic bacteria. The isolates were identified by the test using API 50 CHL medium kit and 16S rDNA partial sequencing. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. When Streptococcus mutans was cultured in the media, the mean weight of formed artificial plaque on the orthodontic wires was $124.4{\pm}30.4\;mg$, whereas being reduced to $5.2{\pm}2.0mg$ and $10.6{\pm}6.6mg$ in the media cultured with Streptococcus mutans and each isolate, respectively (p<0.05) 3. The number of viable cells of Streptococcus mutans was $3.4{\times}10^9$ per ml in the cultured solution, whereas those of Streptococcus mutans in the combined culture with each of isolates were $4.6{\times}10^8\;and\;2.4{\times}10^8$ per ml. 4. The optical density was 1.286 in the supernatant of Fusobacterium nucleatum after vortexing for 30minutes, whereas in the supernatant of combined Fusobacterium nucleatum and each isolate, they were reduced to 0.628 and 0.497, which the percentages of coaggregation between them were 29.4% and 57.8%, respectively 5. The optical density of Fusobacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$, being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusobacterium nucleatum and each isolate. The optical density of Porphyromonas gingivalis precipitate was 1.932 in the culture media, being reduced to 1.170 and 1.266 in the coaggregated precipitates of Porphyromonas gingivalis and each isolate. 6. When two isolates were tested with API 50 CHL medium kit, those were identified Lactobaciallius salivarius and Lactobaciallius delbrueckii subsp. lactis. 7. The similarity values of 16S rDNA sequence between each of isolates and Lactobaciallius salivarius subsp. salicinius were 99.60% and 99.73%, respectively, meaning that isolates were Lactobaciallius salivarius subsp. salicinius. These results indicated that two strains isolated from caries-free children's saliva, which inhibited the formation of artificial plaque and the production of volatile sulfur compounds, were identified as Lactobaciallius salivarius subsp. salicinius.

• Journal of Microbiology
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• v.41 no.3
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• pp.189-195
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• 2003

To enhance the reductive dechlorination of polychlorinated biphenyls (PCBs) under anaerobic conditions, we examined the adjunctive effects of cobalt (Co) and nickel (Ni), which are the central metals of transition-metal cofactors of coenzyme F$\_$430/ and vitamin B$\_$12/, respectively, on the dechlorination of Aroclor 1248. After 32 weeks of incubation, the average numbers of chlorines per biphenyl in culture vials supplemented with 0.2, 0.5, and 1.0 mM of Co reduced from 3.88 to 3.39, 2.92, and 3.28, respectively. However, the numbers of chlorine after supplementing with Ni decreased from 3.88 to 3.43, regardless of the Ni concentrations. The observed congener distribution patterns of all vials with different conditions were similar to the pattern produced by the dechlorination process of H' after 21 weeks of incubation, and these patterns were unchanged up to week 32, except for vials supplemented with 0.5 and 1.0 mM of Co. In vials containing 0.5 mM of Co, meta-rich congeners, such as 25/ 25-,24/25-, and 25/23-chlorobiphenyls (CBPs), which were found as accumulated products of dechlorination in other conditions, were further dechlorinated, and 25/2-, 24/2-, and 2/2-CBPs were concomitantly increased after 32 weeks of incubation. In this case, the congener distribution was similar to the dechlorination pattern of process M. From these results, we suggested that the enrichment of cultures with Co might stimulate the growth of specific populations of meta-dechlorinators, and that populations might promote a change in the dechlorination process from H' to M, which is known to be less effective on the dechlorination of the more highly chlorinated congeners of PCBs.

• Journal of Soil and Groundwater Environment
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• v.13 no.1
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• pp.24-31
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• 2008

Sulfate reducing bacteria (SRB) is universally distributed in the sediment, especially in marine environment. SRB reduce sulfate as electron acceptor to hydrogen sulfide in anaerobic condition. Hydrogen sulfide is reducing agent enhancing the reduction of the organic and inorganic compounds. With SRB, therefore, the degradability of organic contaminants is expected to be enhanced. Ferrous iron reduced from the ferric iron which is mainly present in sediment also renders chlorinated organic compounds to be reduced state. The objectives of this study are: 1) to investigate the reduction of TCE by hydrogen sulfide generated by tht growth of SRB, 2) to estimate the reduction of TCE by ferrous iron generated due to oxidation of hydrogen sulfide, and 3) to illuminate the interaction between SRB and ferrous iron. Mixed bacteria was cultivated from the sludge of the sewage treatment plant. Increasing hydrogen sulfide and decreasing sulfate confirmed the existence of SRB in mixed culture. Although hydrogen sulfide lonely could reduce TCE, the concentration of hydrogen sulfide produced by SRB was not sufficient to reduce TCE directly. With hematite as ferric iron, hydrogen sulfide produced by SRB was consumed to reduce ferric ion to ferrous ion and ferrous iron produced by hydrogen sulfide oxidation decreased the concentration of TCE. Tests with seawater confirmed that the activity of SRB was dependent on the carbon source concentration.

• Korean Journal of Agricultural Science
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• v.34 no.2
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• pp.99-109
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• 2007

Cellulomanas sp. CS 1-1 was studied for its morphological, physiological and biochemical characteristics, together with DNA homology and fatty acid pattern to elucidate its taxonomical position in the species level. Colony morphology of CS1-1 exhibited circular form, opaque, convex, entire edge and pale yellow. Cells were of rod with the size of $0.3{\sim}0.5{\times}0.8{\sim}1.2{\mu}m$, while coryneforms were formed at the early stage of culture. D-ribose, raffinose, rhamnose, acetate, propionate, L-lactate, D-gluconate, aspartate and proline were not utilized as a sole source of carbon, whereas saccharose, arabinose, and amlyose were utilized. Biochemical characteristics of CS1-1 were Gram positive, catalase positive, oxidase negative, nonmotile, facultative anaerobic, mesophilic and G+C content of 74.7 mol %. The major fatty acid and menaquinone were 12-methyltetradecanoic acid(anteiso-$C_{15:1}$) and MK-$9(H_4)$, respectively. These results were correspondent with the characteristics reported for member of the genus Cellulomonas. The strain CS 1-1 exhibited a high level of DNA homology as 70% with C. uda ATCC491, compared to those of 54~59% with C. fimi ATCC 15724, 46~48% with C. biazotea, C. gelida and C. bibula. Finally, strain CS1-1 could be classified as a novel species belongs to C. uda.

• Applied Biological Chemistry
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• v.35 no.3
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• pp.210-217
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• 1992

A photosynthetic bacterium strain KUP-74 producing high amount of S-amino-levulinic acid(ALA) was isolated from soils, which was identified as Rhodocyclus gelatinosus. After 10 days cultivation under anaerobic-light condition at $30^{\circ}C$, 4 Klux and pH 6.8, 5 mg/l of ALA was formed extracellularly. ALA productions were increased up to 8 mg/l and 12 mg/l in cell cultivations either by the addition of 0.5% glycerol (v/v) or 10 mM of glycine and succinic acid, respectively, using Lascelles basal medium eliminated L-glutamic acid. By cultivation in the presence of 30 mM each D,L-glutamic acids and D,L-glutamines the yield of ALA showing a late induction phenomenon was reached the maximum value of 21 mg/l. Different culture times were needed to generate maximum ALA yields by the addition of initial precursors of $C_4$ and $C_5$ pathways in basal medium, as being 107 h and 262 h, respectively. 40 mg/l yield of ALA was observed by cell cultivation with the basal medium containing each 10 mM levulinic acid(LA) and glycine simultaneously.

• Journal of the korean academy of Pediatric Dentistry
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• v.38 no.2
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• pp.146-154
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• 2011

An in vitro study on cariogenic potential of 8 over-the-counter syrups for children was performed. The experimental groups were 8 over-the-counter syrups. The positive control group was 10% sucrose solution, and the negative control group was artificial saliva. The pH of each group was determined. The buffering capacity was measured by the volume of 2 N NaOH adding to equalize the pH of 20 ml of experimental solution to pH 7. The consistency was measured by the time to pass Ostwald pipette for 2 ml of the experimental solution. The experimental solutions were inoculated with S. mutans and cultured in $37^{\circ}C$ anaerobic condition for 48 hours. To estimate acid production, pH of the experimental solutions were measured before and after the culture. The primary teeth specimens were soaked in the experimental solutions for 20 minutes three times a day. Except on those hours the specimens were stored in artificial saliva. After 5 days, the microhardness changes of the specimens were measured. These results show that most of syrup-form medicines for children tend to have cariogenic potential partially in endogenous pH, buffering capacity, consistency, acid production and erosive ability of enamel. For the oral health of children, the alternative sweeteners (ex. xylitol) may be substituted for the cariogenic sweeteners of syrups. Additionally, It may be helpful that the chewable tablet replace liquid or syrup in term of dose form.

• Microbiology and Biotechnology Letters
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• v.25 no.4
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• pp.386-390
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• 1997

The characteristics of alcohol dehydrogenase (ADH, EC 1.1.1.1, alcohol:NAD oxidoreductase) of thermotolerant alcohol-producing yeasts, Saccharomyces cerevisiae RA-74-2 and Kluyveromyces marxianus RA-912, were compared with that of mesophilic S. cerevisiae D, an industrial strain. Under anaerobic culture condition, both S. cerevisiae RA-74-2 and D had similar level of ADH activity at 30$\circ$C, and the activity of S. cerevisiae RA-74-2 at 37$\circ$C was the same level at 30$\circ$C. However, the level of ADH activity of S. cerevisiae D at 37$\circ$C decreased about 70% of that at 30$\circ$C. The level of enzyme activity of K. marxianus RA-912, which showed lower alcohol productivity than S. cerevisiae RA-74-2 and D, was about 43% of those strains at 30$\circ$C, and decreased somewhat at 37$\circ$C. The results showed a good correlation between the alcohol productivities and the level of ADH activities of these strains grown at 30$\circ$C and 37$\circ$C. And the higher heat stability of ADH of S. cerevisiae RA-74-2 than that of S. cerevisiae D seemed to reflect the ability of high temperature fermentation. Despite of its fermentation ability even at 45$\circ$C, however, the ADH of K. marxianus RA-912 showed lower heat stability than that of S. cerevisiae D. Both S. cerevisiae RA-74-2 and D showed similar patterns of two bands of ADH isozyme, and the low band of S. cerevisiae RA-74-2 moved slightly faster than that of S. cerevisiae D. The staining intensity of the bands of S. cerevisiae D at 37$\circ$C was weaker than those at 30$\circ$C. However, S. cerevisiae RA-74-2 showed no differences in total intensity of the bands of 30$\circ$C and 37$\circ$C. As the patterns of cellular proteins and ADH isozyme of K. marxianus RA-912 were different from S. cerevisiae RA-74-2 and D, K. marxianus might have its own characteristic ADH system.