• KSBB Journal
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• v.21 no.6 s.101
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• pp.451-455
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• 2006

An efficient pilot scale (10 ton) three-stage methane fermentation system to digest food waste has been developed in this laboratory. This system consisted of three stages: semianaerobic hydrolysis, anaerobic acidogenesis and strictly anaerobic methanogenesis. From the secondary acidogenesis reactor, a novel strain KA4 responsible for alcohol fermentation was isolated and characterized. The cell was oval and its dimension was $5.5-6.5{\times}3.5-4.5\;{\mu}m$. This strain was identified as Saccharomyces cerevisiae KA4 by 26S rDNA D1/D2 rDNA sequence. Optimal culture temperature was $30-35^{\circ}C$. Cells were tolerant to 5% (v/v) ethanol concentration, however, were inhibited significantly by higher ethanol concentration up to 7%. The strain could grow well up to 50% (w/v) initial glucose concentration in the YM liquid medium, however, optimal concentration for ethanol fermentation was 10%. It could produce ethanol in a broad initial pH range from 4 to 10, and optimal pH was 6. In this condition, the strain converted 10% glucose to 7.4% ethanol during 24 hr, and ethanol yield was estimated to be 2.87 moi EtOH/mol glucose.

• Microbiology and Biotechnology Letters
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• v.13 no.3
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• pp.245-250
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• 1985

In order to increase citric acid productivity. several attempts were made; isolation and characterization of the mutant strain produced citric acid in a high yield, citric acid fermentation in a medium containing relatively higher amount of glucose and citric acid production by the use of semicontinuous ceil recycle system. By the treatment of Candide lipolytica S-109 with NTG, a mutant J-24 was selected as the highest producer of citric acid among the strains formed larger CaCO$_3$ lytic zone. it produced 72g/1 citric acid in 10％ glucose medium. Because mutant J-24 produced 85g/l citric acid and showed 53％ yield in 16％ glucose medium, several factors were adjusted to increase the yield in 16％ glucose medium. 0.8-1.0$\times$10$^{-3}$ P/C ratio, 0.15％ urea, 0.25％ yeast extract were suitable at citric acid production in 16％ glucose medium. Under this condition, J-24 strain produced 93g/l citric acid and showed 58％ yield. Semicontinuous cell recycle system was used to protons the effective production phase, to minimize the product inhibition and to shorten the lag phase. The productivity of semicontinuous cell recycle system was 0.79g/l h while that of batch system was 0.53g/l.h

• Microbiology and Biotechnology Letters
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• v.10 no.1
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• pp.15-26
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• 1982

A potential fungal lipid producer from starch, which was identified as Muror plumbeus, was isolated from natural sources and its optimum cultivation condition for lipid production was investigated. The Mucor plumbeus FRI 0007 showed the highest felt weight and lipid content which were 2.09 $\pm$ 0.24g per 50$m\ell$ of medium and 37.43% on dry weight basis respectively after 20 days incubation on the medium containing 21% of starch as a carbon source. The urea was the best nitrogen source as compared with sodium nitrate, potassium nitrate, magnesium nitrate, ammonium nitrate and ammonium acetate and its optimum concentration was 2.14g/$\ell$, showing 2.39 $\pm$ 0.07 g felt/50$m\ell$ of medium and 50.73% lipid content on dry weight basis after 25 days incubation. Besides the starch as a carbon source and urea as a nitrogen source, the Mucor plumbeus FRI 0007 utilized ZnSO$_4$, MgSO$_4$, NaH$_2$PO$_4$, $K_2$SO$_4$and FeCl$_3$as mineral sources. However, it did not require ail the above 5 minerals in group in-dispensably for its growth and lipid accumulation. The lipid and economic coefficient of Mucor plumbeus FRI 0007 grown on the medium containing 0.44g $K_2$SO$_4$or 5.00g MgSO$_4$/$\ell$solely were 14.96 and 15.37 and 31.12 and 26.10 which was higher than those on the medium containing the above 5 minerals.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.681-692
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• 1996

The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

• Korean Journal of Microbiology
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• v.17 no.3
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• pp.97-130
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• 1979

Many industrial processes those employ bacteria are subjected to phage infestations. In L-glutamic acid fermentions using acetic acid, the phage infestations of the organisms have been recently recognized. In efforts to elucidate the sources of phage contamination involved in the abnormal fermentation, a series of study was conducted to isolate the phages both from the contents of abnormally fermented tanks and the soil or sewage samples from the surroundings of a fermentation factory, to define major charateristics of the phage isolates, and finally to determine the correlation between the phage isolates and temperate phages originating from the miscellaneous bacterial species isolated from the soil or sewage samples. The results are summarized as follows; 1) All phages were isolated from the irregular fermentation tanks and soil or sewage samples, and they were designated as phage PR-1, PR-2, PR-3, PR-4, PR-5, PR-6, and PR-7, in the order of isolation. These PR-series phages were proved to be highly specific for the variant strains of Br. lactofermentum only, namely, phage PR-1 and PR-2 for Br. lactofermentum No. 468-5 and phage PR-3~PR-7 for Br. lactofemrentum No. 2256. By cross-neutralization test, the 7 phagescould be subdivided into 3 groups, i. e., phage PR-I and PR-2 the first, phage PR-3, PR-4, PR-5, PR-6 the second, and the phage PR-7 the third. 2) The 7 phages were virulent under the experimental conditions. They produced plaques with clear and relatively sharp margins without distinct halo. The mean sizes of plaques were 1.5mm in diameter for phage PR-1 and PR-2, and 1. Omm for phages PR-3~PR-7. Double layer technique modified by Hongo and described by Adams, was applied to assay of the PR-series phages. The factors influencing the plaques were as follows;young age cells of host bacteria cultured for 3-6 hours represented the largest number and size, optimum was pH 7.0, incubation temperature was $30^{\circ}C$, and agar concentration and amount of overlayer medium were 0.6% and 0.2ml, respectively. 3) PR-series phages were stable in 0.05M tris buffer and 0.1M ammonium acetate buffer solution. The addition of $5{\times}10^{-3}M$ magnesium ion effectively increased the stability. Thermostability experiments indicated that PR-series phages were stable at the teinperture between $50^{\circ}{\sim}55^{\circ}C$ in nutrient medium, $45^{\circ}{\sim}50^{\circ}C$ in buffer solution. However, the phages mere completely inactivated at 603C and 65$^{\circ}$C within 10 minutes. The phages were stable at the range of pH6~9 in nutrient medium and of pH 8-9 in buffer solution, respectively. Exposure of the phages to UV for 25, 60 and 100 seconds resulted in the complete loss of infectivily, respectively. 4) Electron microscopy showed that PR-series phage particles exhibited rather similar morphology, differing in the size All of PR-series phages had a multilateral head and had a simple long tiil about three to five times long as compared with head. By the size, phage PR-1 and PR-2, PR-3, PR-4, PR-5, and PR-6 and PR-7 were classified into same groups, respectively. The head and tail size of phage PR-1, PR-5, PR-5(T) and PR-7 were 85nm, 74nm and 235nm and 350mm, and 72nm and 210nm, respectively. 5) Nucleic acids of PR-series phages were double stranded DNA. The G+C contents of phage PR-1, PR-5 and PR-7 were 56.1, 52.9 and 53.7, respectively. The values of G+C contents derived from the $T_m$ were in agreement with the chemically determined values. 6) PR-series phages effectively adsorbed on their host bacteria at the rate of more than 90% during 5 min. K value for phage PR-1, PR-5 and PR-7 were calculated to be $6{\times}10^9 ml$ per minute, respectiveky. The pH of the medium did effect adsorption rate, but both temperature and age of host cells did not. Generally, optimum adsorption condition of phages seemed to be almost same as optimum growth conditions of host bacteria. 7) In one-step growth experiments, the latent periods at $30^{\circ}C$ for PR-1, and PR-7 were about 70, 50 and 55 min, respectively. The corresponding average burst size was 200, 70 and 90, respectively. Lpsis period according to the multiplicity of infection and a phage series. In case of m. o. i. 100, strain No. 2256 (PR-5) and No. 468-5(PR-1) failed to grow and turbidity decreased after 50 and 70min, respectively. 8) In the lysate of a plaque purified phage PR-5 infected bacteria, there observed 2 types ofphage particles, i. e., phage PR-5 and PR-5 (T) of similar morphology but differing at the length of phage tail, and phage tail like particles. The phage taillike particles could be divided into 4 types by the length. Induction experiments of Br. lactofermentum with UV irradiation, mitomycin C or bacitracin treatment produced neither phage PR-5 (T) or phage tail-like particles. 9) No lysis occured when the growth of 7 strains of miscellaneous bacteria, isolated from soil and sewage samples, were inoculated with either phage PR-5 (T) or phage tail-like particles the inoculation of phage PR-5 pellet resulted in the growth inhibition of the orgainsms in the spot test. The lysates obtained from 3 miscellaneous soil derived bacteria following mitomycin C treatment the growth of Br. lactofermentum, but did not lyze the bacterium.

• Korean Journal of Veterinary Research
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• v.50 no.3
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• pp.213-220
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• 2010

Salmonella (S.) Enterica infection ranks among the most common food borne bacterial infections worldwide. Although there are six subspecies of S. Enterica, the vast majority of human and animal infections are caused by strains belonging to subspecies 1 serovar Typhimurium and Enteritidis. Recent reports on antibiotic resistance of Salmonella spp. are rising steadily. The increasing problem of antibiotic resistance has rekindled interest in bacteriophage to therapy. Therefore, we investigated the efficacy of bacteriophage in S. enterica serovar Enteritidis infected mice and pigs by measuring of body condition, body weight, bacterial colonization and weight of organs based on the in vitro analysis. In vitro experiment, phage cultured with S. Enteritidis showed clear lysis pattern, the plaque forming unit (PFU) of our phage culture was $1.5{\times}10^{11}PFU/mL$, and phage showed its maximum activity at 4 h post inoculation. In mouse experiment, there was no significant difference among experimental groups in the general body conditions and body weight of mice. However, there was difference in weight of liver and spleen depending on the experimental group (p < 0.05). The weight of liver and spleen were reduced by the phage treatment. Also bacterial colonization in spleen and liver were significantly reduced by the phage treatment. In pig experiment, the general body conditions and body temperature exhibited not much difference among the pigs except few pigs in group 3 which showed poor body conditions. From the feces in each group, we could isolate the S. Enteritidis only from group 3. Bacterial enrichment culture was necessary for isolating the bacteria from 5 dpi and 10 dpi, however direct isolation was possible from 15 dpi feces. In phage treated group, postmortem lesion was better than non-phage treated group. Recently, antibiotic resistance concerns on the food-borne bacterial pathogens have been increasing because of the wide spread of the antibiotics resistance genes. This concern is widely transmitted to the human related public health. As one of the alternative treatments on the bacterial pathogens, attempt using phages have been made to control the bacterial diseases. The positive possibility of the trail using phage was observed to control the S. enterica serovar Enteritidis in this study even though the further analysis has been remained.

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

• The Journal of Engineering Geology
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• v.31 no.4
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• pp.659-669
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• 2021

The purpose of this study is to elucidate the relationship between occurrence of natural radioactive materials such as ²³⁸U and ²²²Rn and original mixing ratio of helium isotope of groundwater from various geology, and to suggest the underground aquifer environment from helium original mixing data. 9 groundwater samples were collected from five study areas, and ²³⁸U, Rn-222 and helium isotope were analyzed. A high ²³⁸U content of the range of 218~477 ㎍ /L in the groundwater occurs in the twomica granite. ⁴He air-crust mixing ratio and the Rn-222 content show a rough relation, that is, Rn-222 content increases according to the increase of ⁴He crust mixing ratio. Because of helium and radon are an inert gas, their behavior in underground environment is assumed as an analogous. The ²³⁸U content and He isotope in groundwater does not show any distinct correlation. The groundwater can be classified as three groups (air, air-crust mixing, crust-mantle mixing origin) on the diagram of ³He/⁴He vs ⁴He/²⁰Ne, which is composed of original mixing line from air-crust-mantle end members. This original mixing of helium can provide the information of underground aquifer characteristic such as the connection with surface environment or isolation condition from air environment.

• The Korean Journal of Food And Nutrition
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• v.14 no.6
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• pp.548-561
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• 2001

Listeria spp. from sea water, fishes and shellfishes have been troubled in many countries. So we exam ined its distribution rates, biochemical characteristics of a separated strain, growth curve of pH at set times to 4 species of standard strain, and yes or no of growth inhibition for precautionary measure of food poisoning by L. monocytogenes, garlic, mustard, wasabi, and green tea extracts including sensitivity of antibiotics 10 species. As its results, check numbers of its positivity to Listeria spp. were 32 species in total examination body 200 species, and its isolation rates were 16%, L. innocua was 14.0%, L. monocytogenes 1.0%, and L. seeligeri 1.0% by the strain species. All the standard strain of 4 species showed growth inhibition bellow pH 3.0, its pH conditions of the optimum growth at 7.0∼8.0, and its growth was more active in alkali co]tuition than in acid condition. Its growth inhibition examination by garlic extracts had an the worst effects with O.D values of 0.078∼0.210. But the case of mustard and wasabi had weakened effect, and the case of green tea had some effect as the time went by. The results of sensitivity examination of antibiotics 10 species were as fellows. L. innocua of the 16 cases showed sensitivity of 100% in all 5 species, Ampicillin, etc, and Ciprofloxacin showed sensitivity of 43.7% and gentamicin, 93.7%. But tetracycline showed tolerance of 31.3% , cefotaxine. 75%, nalidixic acid, 100%. L. monocytogenes of the 6 cases showed sensitivity of 100% in all 6 species, ciprofloxacin, etc.

• KSBB Journal
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• v.25 no.2
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• pp.155-166
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• 2010

Studies on the production of cell-wall bound innerpolysaccharides (IPS) (soluble ${\beta}$-D-glucan) have been performed by use of suspended myelial cultures of Inonotus obliquus. This product has promising potentials as an effective antidiabetic as well as an immunostimulating agents. As a first step to enhanced production of IPS, Intensive strain improvement programs were carried out by obtaining a large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because about fivefold higher amount of protoplasts ($2.3{\times}10^6$ protoplasts/mL) could be recovered with relatively high regeneration rates of $10^{-2}{\sim}10^{-3}$ by applying a modified filtration method, as compared to the previously used trapping method. A basic protocol necessary for UV-mutation of the protoplasts was also developed, resulting in several overproducing variants with good fermentation properties. Since the amount of IPS extracted from the mycelial cell walls of I. obliquus turned out to be almost constant per g DCW, increase in cell mass was considered the most important factor for the enhancement in IPS production. Therefore, attempts were made to screen mutant cells showing rapid mycelial growth rate in the final suspended cultures. Notably, the mutant strains showing an active cellgrowth in the preceding solid growth cultures were observed to produce higher amount of IPS in the suspended fermentations as well. A striking mutant, OBLQ756-15-5 strain, obtained from the survivors of a harsh UV-treated condition (97% death rate) was found to stably produce as high cell mass as 22 g DCW/L in the final fermentations. Currently, this strain is being tested for development of a scaled-up fermentation process for mass production of IPS.