• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.519-526
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• 1992

The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

• Korean Journal of Veterinary Research
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• v.43 no.1
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• pp.11-24
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• 2003

This experimental studies was to investigate the location of PNS and CNS labeled neurons following injection of 2% WGA-HRP and pseudorabies virus (PRY), Bartha strain, into the ductus deferens of rats. After survival times 4-5 days following injection of 2% WGA-HRP and PRV, the rats were perfused, and their brain, spinal cord, sympathetic ganglia and spinal ganglia were frozen sectioned ($30{\mu}m$). These sections were stained by HRP histochemical and PRY inummohistochemical staining methods, and observed with light microscope. The results were as follows ; 1. The location of sympathetic ganglia projecting to the ductus deferens were observed in pelvic ganglion, inferior mesenteric ganglion and L1-6 lwnbar sympathetic ganglia. 2. The location of spinal ganglia projecting to the ductus deferens were observed in T13-L6 spinal ganglia. 3. The PRY labeled neurons projecting to the ductus deferens were observed in lateral spinal nucleus, lamina I, II and X of cervical segments. In thoracic segments, PRY labeled neurons were observed in dorsomedial part of lamina I, II and III, and dorsolateral part of lamina IV and V. Densely labeled neurons were observed in intermediolateral nucleus. In first lumbar segment, labeled neurons were observed in intermediolateral nucleus and dorsal commisural nucleus. In sixth lumbar segment and sacral segments, dense labeled neurons were observed in sacral parasympathetic nuc., lamina IX and X. 4. In the medulla oblongata, PRV labeled neurons projecting to the ductus deferens were observed in the trigeminal spinal nuc., A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nuc., rostroventrolateral reticular nuc., area postrema, nuc. tractus solitarius, raphe obscurus nuc., raphe pallidus nuc., raphe magnus nuc., parapyramidal nuc., lateral reticular nuc., gigantocellular reticular nuc.. 5. In the pons, PRV labeled neurons projecting to the ductus deferens were ohserved in parabrachial nuc., Kolliker-Fuse nuc., locus cooruleus, subcooruleus nuc. and AS noradrenalin cells. 6. In midbrain, PRV labeled neurons projecting to the ductus deferens were observed in periaqueductal gray substance, substantia nigra and dorsal raphe nuc.. 7. In the diencephalon, PRV labeled neurons projecting to the ductus deferens were observed in paraventricular hypahalamic nuc., lateral hypothalamic nuc., retrochiasmatic nuc. and ventromedial hypothalamic nuc.. 8. In cerebrum, PRV labeled neurons projecting to the ductus deferens were observed in area 1 of parietal cortex. These results suggest that WGA-HRP labeled neurons of the spinal cord projecting to the rat ductus deferens might be the first-order neurons related to the viscero-somatic sensory and sympathetic postganglionic neurons, and PRV labeled neurons of the brain and spinal cord may be the second and third-order neurons response to the movement of smooth muscles in ductus deferens. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitaing the internal environment. These observations provide evidence for previously unknown projections from ductus deferens to spinal cord and brain which may be play an important neuroanatornical basic evidence in the regulation of ductus deferens function.

• Korean Journal of Soil Science and Fertilizer
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• v.24 no.1
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• pp.61-68
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• 1991

Thirty Bradyrhizobium japonicum isolates (10 strains per each soil) from 1 uncultivated [Sangnam(Soil 1), Milyang]- and 2 cultivated [Dong(Soil 2)and Chinbuk(Soil 3), Changweon] upland soils in Yeongnam area were evaluated on their symbiotic effectiveness to soybean [Glycin max (L.)] cv. Korean Jangbaekkong and American Clark and examined on their serological diversity. The results obtained were summarized as follows : 1. On symbiotic effectiveness of B. japonicum with plant genotypes, isolates showed a relatively high value of nodule mass in Jangbaekkong cv. and of shoot dry weight and total nitrogen in Clark cv. demonstrating the order of Soil 1> Soil 2> Soil 3 isolates. 2. Among 30 B. japonicum isolates, YCK 141 showed the best effectiveness on mean nitrogen fixation of two cultivars. 3. Thirty indigenous B. japonicum showed 6 types of serological diversities in the immunoblot analysis which were present in various proportions at Soil 2(5) and Soil 3(5) except Soil 1 where all isolates fell into the YCK 117 serogroup. And their distribution order was serotype YCK 117( 12 strains) > USDA 1l0(5strains), USDA 123(5 strains) > YCK 150(4 strains) > YCK 141(3 strains) > YCK 226(1 strain). 4. Especially, 10 isolates from Soil 1, an uncultivated orchard, showed a very homologous pattern in not only effectiveness but serological distribution. It seemed to indicate that the isolates were typically affected by numerous physical and environmental factors of the soil.

• Korean Journal of Soil Science and Fertilizer
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• v.29 no.1
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• pp.74-80
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• 1996

This experiment was conducted to obtain the basis of degradation of remaining agricultural chemicals accumulated in upland soils of Honam district in Korea. The population. relative growth rate(RGR). chlorothalonil(TPN)-degradation ability and bacterialogical characteristics of TPN resistant strains were investigated in TPN levels of 0, 25, 50, 100 and $500{\mu}l/ml$ compared with Mancozeb. A number of TPN-resistant bacteria were differ in the area of examined and were decreased with higher levels of TPN. The resistance of bacteria was stronger in TPN than Mancozeb but the resistance of fungi was vise versa. RGR of bacteria in the culture was the highest at the level of $50{\mu}l/ml$ and the lowest in $500{\mu}l/ml$ of TPN. TPN-degradation ability of bacteda isolated in various TPN levels was varied : only 8 percentage of bacteria showed 75 percentage or more degradation ability. The higher the concentration in TPN resistance, the larger the number of strains carried great ability to decompose pesticide residues. The strains having higher decomposition ability was rod-shapes cells and senstive to heat. Analyses of the indol production, methyl red, and V-P test have given similar results, with negative reaction in all these strain, while the other biochemical characteristics were differ in the strains. Based on these, these strains might be classified into Pseudomonas sp., Corynebacterium sp., Acinetobacter sp. and Moraxcella sp.

• 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 Microbiology
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• v.48 no.2
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• pp.109-115
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• 2012

Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.

• Korean journal of applied entomology
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• v.26 no.4 s.73
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• pp.229-237
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• 1987

In order to study the biological control of soil-borne disease of sesame, antagonistic isolates of Trichoderma , Bacillus sand streptomyces to Fusarium oxysporum and Rhizoctonia solani were isolated from the rhizosphere soils of sesame plants and some other habitats. Out of the isolates of microorganisms collected a strain of Trichoderma viride was selected as a biological control agent for the study and its effect on the control of damping-off and the seedling growth of sesame was investigated. The results obtained are as follows: 26 percents of Bacillus spp. isolated from the rhizosphere soil of sesame plants showed antagonism to two pathogenic fungi. Important species were B. Subtilis and B. polymyxa. Streptomyces species isolated from the rhizosphere soils of sesame lysed the cell wall of hyphae and conidia of F. oxysporum and reduced conspicuously the formation of macroconidia and chlamydospores of the fungus. 84 percents of Trichoderma spp. isolated from the rhizosphere soil of sesame plants were antagonistic to F. oxysporum and 60 percents of the isolates were antagonistic to both F. oxysporum and R. solani. Trichoderma viride TV-192 selected from antagonistic isolates of Trichoderma spp. was highly antagonistic to F. oxysporum and soil treatment with the isolate reduced notably damping-off of sesame. T. viride TV-192 showed better growth in crushed rice straw, barley straw and sawdust media than F. oxysporum. Sawdust was selective for the growth of T. viride. Supplementation of wheat bran and mixtures of wheat bran and sawdust inoculated with T. viride TV-192 in the soil reduced remarkably damping-off of sesame by F. oxysporum but high density of the fungus TV-192 caused the inhibition of seed germination and seedling growth of sesame. Inhibitory effects of Trichoderma species on seed germination and seedling growth of sesame were different according to the isolates of the fungus. Normal sesame seedlings on the bed treated with the fungus showed better growth than not treated seedlings.

• Korean Journal of Agricultural Science
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• v.6 no.2
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• pp.205-212
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• 1979

Red pepper paste were prepared by using various raw materials such as rice, glutinous rice, polished barley, polished wheat and corn powder, and their chemical compositions including reducing sugar, amino-N and ethanol were analyzed during the period of aging in order to elucidate effects of these starchy materials on the compositions and qualities of the products. The results obtained are summarized as follows: 1) Koji of these materials were manufactured respectively by inoculating a strain of Asporyzae and their activities of protease and saccharifying amylase were determined. Wheat koji was found to have the highest level of protease activity among the koji, while glutinous rice koji had the strongest amylase activity. 2) Contents of moisture, crude protein, crude fat, and sodium chloride in the red pepper pastes were not changed significantly, however total sugar content was decreased during the period of aging. 3) After 60 days of aging, the highest amount (160mg%) amino nitrogen was detected in the red pepper paste of polished wheat, but higher levels of reducing sugar and ethanol contents were detected in that of glutinous rice. 4) Amino acids in the products were analyzed after 60 days of aging: a) Total 17 amino acids were detected. b) Free amino acid ratio to total amino acid content was approximately 35 percent in average. c) Free glutamic acid was higher than any other free amino acid In the amount and free ratio, but free methionine was lower. d) Lysine content was relatively high, while histidine and glycine were detected in trace. 5) Sensory tests including color, odor and taste on the products resulted in the highest mark for the red pepper paste of glutinous rice but in the lowest mark for that of corn powder.

• Journal of Life Science
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• v.20 no.9
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• pp.1373-1377
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• 2010

Coastal sand-dune plants can survive very effectively with the help of various microbes, especially ecto- and endomycorrihizae. Penicillium citrinum KACC43900 was screened according to growth promoting activity on sand-dune plants Calystegia soldanella and Ischaemum anthephoroides. In this study, coastal sand-dune plants were treated with a fungal culture filtrate of endophytic fungus P. citrinum KACC43900 to confirm the promotion of plant growth. C. soldanella and I. anthephoroides were used as representative coastal sand-dune plants, and their shoot length, plant length, and dry weight were analyzed. The shoot lengths of control C. soldanella and I. anthephoroides were 8.60 cm and 8.12 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 16.30 cm and 10.56 cm, respectively. The plant lengths of control C. soldanella and I. anthephoroides were 14.90 cm and 14.80 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 24.06 cm and 17.06 cm, respectively. The dry weight of C. soldanella and I. anthephoroides treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 0.163 g and 0.032 g, respectively. It was shown that the growth of shoots in C. soldanella and I. anthephoroides was promoted, 89.54% and 29.60%, by culture filtrate of P. citrinum KACC43900.

• Tuberculosis and Respiratory Diseases
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• v.47 no.2
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• pp.161-171
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• 1999

Background: Interleukin-12 (IL-12) can induce antitumor effects in vivo. This antitumor effect is associated with T cell infiltration but the effect of IL-12 on the steps of T cell migration into the tumor tissue has not been fully elucidated. This study focused on the effect of IL-12 on the tumor growth and the metastasis and on the expression of E-selectin, an adhesion molecule which is activated endothelial specific in its expression. In addition, we studied whether the expression of E-selectin is associated with the TNF-$\alpha$, a cytokine that its production is increased by IL-12 and has functions inducing a variety of adhesion molecules. Methods: Mice of C57BL/6 strain were injected with Lewis lung cancer cells followed by either IL-12, TNF-$\alpha$, or normal saline by intraperitoneal route. Twenty eight days after tumor cell inoculation, metastatic nodules of lung were enumerated and immunohistochemical staining of the subcutaneous tumors were performed with monoclonal antibodies to CD4, CD8, CD16, and E-selectin. In IL-12 treated mice, the subcutaneously implanted Lewis lung tumors were decreased in size and the metastases were also decreased in number compared to control mice. On tumor tissues, increased infiltration of CD4+, CD8+, and CD16+ cells were oberved in IL-12 treated mice compared to control mice. In control mice, E-selectin was absent on tumor vessels, but the expression of E-selectin was increased on tumor vessels of IL-12 treated mice. Administration of TNF-$\alpha$ increased not only the expression of E-selectin but also infiltrations of CD4+, CD8+, and CD16+ cells on tumor tissues. Conclusions: These results demonstrate that IL-12 inhibits tumor growth and metastases through infiltrations of inflammatory cells in mouse model of Lewis lung carcinoma and E-selectin may playa role in inflammatory cell recruitment on tumor tissue following IL-12 administration. Also, TNF-$\alpha$ may have a role as a mediator responsible for the IL-12 induced expression of E-selectin.