• Proceedings of the Korean Society of Life Science Conference
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• 2001.06a
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• pp.67-86
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• 2001

A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

• Journal of Life Science
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• v.23 no.2
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• pp.273-281
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• 2013

The purpose of this research was to investigate the production and characterization of alkaline protease from Micrococcus sp. PS-1 newly isolated from seawater. Micrococcus sp. PS-1 was grown in Luria-Bertani (LB) medium. Its optimal temperature and pH for growth were $30^{\circ}C$ and 7.0, respectively. The effect of nitrogen sources was investigated on optimal enzyme production. A high level of alkaline protease production occurred in LB broth containing 2% skimmed milk. The protease was purified in a 3-step procedure involving ultrafiltration, acetone precipitation, and dialysis. The procedure yielded a 16.43-purification fold, with a yield of 54.25%. SDS-PAGE showed that the enzyme had molecular weights of 35.0 and 37.5 kDa. Its maximum protease activity was exhibited at pH 9.0 and $37^{\circ}C$, and its activity was stable at pH 8.0-11.0 and $25-37^{\circ}C$. The protease activity was strongly inhibited by PMSF, EDTA, and EGTA. Taken together, the results demonstrate that the protease enzyme from Micrococcus sp. PS-1 probably belongs to a subclass of alkaline metallo-serine proteases.

• Korean Journal of Organic Agriculture
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• v.7 no.2
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• pp.89-97
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• 1999

Through manifestation of the principles of commodity production spatial sharing of labour can be observed in the agriculture as well as in cattle production. Better adjustment of the production structure to the environment factors brings higher yields and more effective production. In being able to maximize the profit the entrepreneurs opt for producing output that closely matches to their featuring conditions. In contrary to the relatively high "mobility" of crop production animal husbandry and within this cattle production - as known - is strictly chained to forage production. On the basis of our economic research and as a result of multivariable analysis(factor analysis) it can be concluded that two variable groups(factors) are highly dominant in organizing profitable cattle production. First of them is the crop site factor (indicated by gold crown value), the second is the forage production feature(forage and grassland area and the yield of them). During recent years the weight of environmental factors suffered from devaluation. As a result of the central economic administration differentiating effects were suppressed and the chances of equalizing concepts strengthened. The outcome can be observed even today. In the regions, for example, being predominantly suitable for grass and forage cropping the milk and slaughter cattle production decreased. The same is ture for com and pig production regions. Unexploitment of local environmental features can be observed mainly in grassland management. Branches being potential user of grasslands hardly take them into consideration. Main method of rational use of grasslands is pasturing. Presence of pastures and the usage of them through cattle production is highly important not only for profitable production but also for maintaining ecological stability.

• Korean Journal of Organic Agriculture
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• v.23 no.3
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• pp.469-480
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• 2015

The study was conducted on the effects of time of defoliation on fruit quality of pear (Pyrus pyrifolia Nakai) trees, managing with low-pesticides, and regrowth of stem cuttings in vitro. Treatments included for 40% of uniform defoliation at early-August, end-August, and early-September, as well as control (no defoliation). Defoliation at early-September and control increased growth of water sprouts as well as concentrations of carbohydrates, total nitrogen, and free sugar in one-year old shoots. Defoliation at early-September and control increased fruit yield and mean fruit weight, with high soluble solids content and fruit surface color of $a^*$ observed for both defoliation at end-August and early-September. Defoliation at early-August increased rates of electrolyte leakage in stem cuttings at $-18^{\circ}C$ in vitro. There were no significantly different for germination rates of the cuttings between the treatments at -18 and $-21^{\circ}C$ in vitro, with the highest germination of the cuttings observed for defoliation at early-September and control at $-27^{\circ}C$. Therefore, orchard management should be performed to be minimized for defoliation of the spur leaves until end-August, causing from precipitation and pests.

• Korean Journal of Organic Agriculture
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• v.23 no.3
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• pp.523-534
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• 2015

Twenty four Holstein steers (average body weight $714{\pm}13.60kg$) were used in this experiment to determine the effect of supplementing of microbial culture and coated vitamin-C on growth performances and carcass characteristics in finishing Holstein steers. Holstein steers were randomly assigned to feeding groups of control group (Con, 12 kg of basal diet/head/day), microbial culture group (MC, 12 kg of basal diet + 30 g of microbial culture/head/day) and coated vitamin-C group (CVC, 12 kg of basal diet + 10 g of coated vitamin-C/head/day). MC and CVC groups were higher in ADG compared to control (P<0.05). FCR was also lower in MC and CVC groups than control group (P<0.05). Back fat thickness, rib-eye area, marbling score, meat color and yield index were not changed by supplementing microbial culture and coated vitamin-C. MC group was higher for maturity compared to control and CVC group (P<0.05). CVC group was higher for fat color compared to control and MC group (P<0.05). Based on the results obtained from the current study, supplementation of microbial culture and coated vitamin-C as an alternative to antibiotic might increase growth performances and enhance carcass characteristics in finishing Holstein steers. However, more studies are needed to find out the optimum supplementing period of microbial culture or coated vitamin-C for high quality meat production from Holstein steers.

• Korean Journal of Organic Agriculture
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• v.19 no.3
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• pp.385-396
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• 2011

This study was carried out to develop environmental friendly control for major diseases and pests on Boxthorn (Lycium chinense Mill.). Outbreak of Eighteen diseases and pests were found at the Boxthorn organic yards in Chung-nam province. Among them Powdery mildew (Erysiphe polygoni de Cand.), Hypophyllous mold (Pseudocercospora chengtuensis (Tai)), Western flower trips (Frankliniella occidentalis (Pergande)), Green peach aphid (Myzus pericae (Sulzer)) and Corn earworm (Helicoverpa armigera) needed to be controled by environmental friendly methods for high fruit yield of organic Boxthorn. In summer(Jun) test Bacilus subtilis QST 713 wettable powder and Sulfur wettable powder were effective and in autumn (Sep.) test Sulfur, Copper hydroxide and Paraffinic oil were relatively effective in Powdery mildew. In Hypophyllous mold control test Paraffinic oil and Bacilus subtilis GB - 0365 were effective with above 70% control value. And it was possible to control Western flower trips by natural enemy (Orius laevigatus) by 80% control value. Corn earworm was possible to control by Bacillus thuringiensis subsp. aizawai GB413 flowable and Bacillus thuringiensis aizawa 0423 wettable powder application above 70%. And Green peach aphid was controllable with environmental friendly materials, such as, Bacillus subtilis (Seoncho), Bacillus subtilis (Jinsami) above 80% and Ginkgo nut extract above 70% control value.

• Horticultural Science & Technology
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• v.28 no.3
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• pp.409-414
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• 2010

The first drainage time in a day was controlled for precise irrigation management with low consumption of nutrient solution in tomato perlite bag culture system by measuring water level of drained water in drainage catchment part. This method automatically adjusted the irrigation time under any condition of light, temperature and humidity, resulting in stable water content in substrates. However, it was difficult to keep the time consistent as they were set. It drained with the deviation of 20 min in the treatment in which the first drainage time was set at 10:00 and 50 min in the treatment set at 10:30. The first drainage time was not constant, but the drain occurred stably before noon in the treatment of which irrigation frequency was longer than 30 min. The drainage ratio was better balanced in all the treatments using drainage level sensors than the treatment using time clock for irrigation control. High water and fertilizer efficiencies were obtained. Although the growth, total yield and sugar content were not significantly different between the treatments, fruit weight was higher in the treatments using drainage level sensors than that using timer.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.851-856
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• 2005

Although valuable microbes have been isolated from the soil for the various productions of useful components, the microbes which can be cultivated in the laboratory are only $0.1-1\%$ of all microbes. To solve this problem, the study has recently been tried for making the valuable components from the environment by directly separating unculturable micrbial DNA in the soil. But it is known that humic acid originated from the soil interrupts various restriction enzymes and molecular biological process. Thus, in order to prevent these problems, this study modified the method separated soil DNA with phenol, CTAB and PEG. In order to compare the degree of purity for each DNA and the molecular biological application process, $A_{260}/A_{280}$ ratio, restriction enzymes, and PCR were performed. In case of DNA by the modified method, total yield of DNA was lower but $A_{260}/A_{280}$ ratio was higher than the previously reported methods. It was confirmed that the degree of purity is improved by the modified method. But it was not cut off by all kinds of tested restriction enzymes because of the operation of a very small amount of interrupting substances. When PCR was operated with each diluted DNA in different concentrations and GAPDH primer, the DNA by the modified method could be processed for PCR in the concentration of 100 times higher than by the previously reported separation method. Therefore, this experiment can find out the possibility of utilization for the unknown substances by effectively removing the harmful materials including humic acid and help establishing metagenomic DNA library from the soil DNA having the high degree of purity.

• Journal of The Korean Society of Grassland and Forage Science
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• v.23 no.3
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• pp.193-202
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• 2003

This study was conducted to investigate the effects of the type and application level of swine manure on herbage productivity, efficiency of nitrogen utilization and environmental pollution by the leaching of nitrogen compounds in mixed grassland. The field experiment was carried out on established grassland sward growing on silt clay loam soil. Main plots were the types of swine manure, such as swine manure fermented with sawdust(SMFWS), swine manure fermented without sawdust(SMF) and swine slurry(SS), and mineral fertilizer(Urea). Subplots were the application levels of swine manure, such as 100, 200 and 400 kgN/ha, The results obtained were summarized as follows; 1. Herbage productivity, nutritive value and nitrogen yields were the highest with mineral fertilizer and followed by swine slurry(SS), and the lowest with swine manure fermented with sawdust(SMFS). As compared with dry matter(DM) yield by application of mineral fertilizer(100%), the DM percents of SMFWS, SMFS and SS were 90.6, 80,9 and 76.8%, respectively. 2. Organic matter(OM) contents of the soil were increased by the applications of swine manure. OM contents were the high set with SMFS and the lowest with mineral fertilizer. 3. The amounts of nitrate leaching by types and application levels of swine manure were the highest in the end of Aug. and early of Sep. during the seasons and ranged from 10 to 25ppm.

• Korean Journal of Food Science and Technology
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• v.48 no.5
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• pp.461-465
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• 2016

In order to prevent the blackening and texture softening of heated shrimp, the pH was adjusted by soaking shrimps in acidic and alkali solutions, and their qualities were evaluated. The lightness of shrimps pretreated with 0.2% (w/w) citric acid and 0.05% (w/w) ascorbic acid solution increased by 20% compared to that of the control. The strength of mechanical hardness of shrimps soaked in acetic acid and phosphate solution (pH 6.0) was significantly higher ($1209g_f$) compared to that of untreated shrimp ($801g_f$; p<0.05), and the overall preference of texture was 0.4 points higher than that of the control in the descriptive sensory evaluation (p<0.05). In contrast, soaking in solution of pH 8 exhibited a weak texture hardening effect ($855g_f$). Additionally, the hardness of the heated shrimp after soaking at an adjusted pH of 4.0 increased to $4046g_f$, but the yield based on weight decreased to 38% compared to that of untreated shrimp (70%; p<0.05).