• Korean Journal of Soil Science and Fertilizer
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• v.14 no.2
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• pp.70-75
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• 1981

The ammonia volatilization from two different soils, an acidic normal soil and a neutral tidal soil applied with different nitrogen sources was investigated through a laboratory incubation experiment conducted at about $30^{\circ}C$ for 18 days. Results obtained were summerized as follows; 1. The ammonia volatilizat ion was increased by the urea application that increased soil pH. 2. Ammonium sulfate and ammonium chloride did not raise reduced soil pH over 7.30 and showed little ammonia volatilization keeping the $pK_b$ value of 4.72-3 3. An organic fertilizer (Miweon Co. made) raised pH of the tidal land soil little more than ammonium sulfate or ammonium chloride ; however, it did not increase the ammonia volatilization as much as from other fertilizer treatment plots of the same pH, which may mean that the organic fertilizer is effective in reducing ammonia volatilization. 4. It seemed that easier volatilization of ammonia from urea may occor in ordinary soil low in original pH than from tidal soil by the application of urea which may mean that if the pH of soils are the same, greater volatilization would result from the former than the latter. 5. Application of raw straw to tidal soil lowed pH and reduced ammonia volatilization.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.333-340
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• 2016

An alkaline protease was purified and characterized from an alkalophilic microorganism, Bacillus sp. DK1122, isolated from soil in central Korea. The optimum temperature and pH for the growth of the producer strain were 40℃ and pH 9.0, respectively. The protease was produced aerobically at 40℃ after 24 h incubation in modified Horikoshi I medium (pH 9.0) containing 0.5% (w/v) glucose, 0.8% (w/v) yeast extract, 0.5% (w/v) polypeptone, 0.1% (w/v) K₂HPO₄, 0.02% (w/v) MgSO₄·7H₂O, 1% (w/v) Na₂CO₃, and 3% (w/v) NaCl. The alkaline protease was purified by 70% ammonium sulfate precipitation of the culture supernatant of Bacillus sp. DK1122, followed by CM-Sepharose chromatography. The molecular weight of the enzyme was estimated to be 27 kDa on the basis of SDS-PAGE. The optimum temperature and pH for the protease activity were 60℃ and pH 9.0, respectively. Addition of CaCl₂ increased the thermal stability of the purified protease, where 90% of protease activity was retained at 60℃ for up to 3 h. Consequently, it is expected that the alkaline protease from this study, exhibiting stability at pH 7–9 and 60℃, may be promising for application in the food and detergent industries.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.915-921
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• 2001

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.6
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• pp.759-764
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• 2001

This study was carried out to investigate the effect of activation timing, cell cycle and passage on the development of embryos produced by cumulus cell nuclear transfer in Hanwoo (Korean cattle). Nuclear donor cumulus cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at $38.5^{\circ}C$ in a humidified atmosphere of 5% $CO_2$ in air. The 1~6 passages of serum deprived or actively dividing cumulus cells were isolated and used as donor cells. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. One pulse of 180 volts for $15{\mu}s$ was applied to induce the fusion between karyoplast and cytoplast. The activation was done before or after the fusion. To activate, oocytes were treated with $10{\mu}M$ calcium ionophore for 5 min immediately followed by 2 mM 6-dimethylaminopurine for 3 h. The nuclear transfer embryos were cultured in $500{\mu}l$ of modified CRlaa supplemented with 3 mg/ml BSA in four well dish covered with mineral oil. After 3 days culture, culture medium was changed into modified CRlaa medium containing 1.5 mg/ml BSA and 5% FBS for 4 days. The incubation environment was 5% $CO_2$, 5% $O_2$, 90% $N_2$ at $38.5^{\circ}C$. There was no blastocyst formation when the nuclear transfer embryos were activated before the fusion, whereas, 29.9% of blastocyst formation was shown when the nuclear transfer embryos were activated after the fusion. When serum deprived and actively dividing cumulus cells were used as nuclear donor cells, the developmental rates to blastocyst were 38.5% and 40.6%, respectively. There was no significant difference between serum deprived and actively dividing cells in the developmental rates. The developmental rates to blastocyst according to 1~6 passages were 37.5~44.4%. However, there were no significant differences among passages. These results indicate that 1~6 passage cumulus cell irrespective of cell cycle could support development of nuclear transfer embryos activated after the fusion.

• Microbiology and Biotechnology Letters
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• v.45 no.2
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• pp.162-167
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• 2017

For the production of bioethanol by the synchronous saccharification and fermentation (SSF) process, bio-capsule formation was attempted. Many saccharifying fungal strains and fermentative yeast strains were first screened. Aspergillus sp. BCNU 6200, Penicillium sp. BCNU 6201, and P. chrysogenum KACC 44363 were found to be excellent producers of saccharifying enzymes such as ${\alpha}$-amylase and glucoamylase. Saccharomyces cerevisiae IFO-M-07 showed the highest ethanol productivity among the tested strains. Secondly, we determined the optimal conditions for pellet formation, and those for bio-capsule formation. All the tested fungal strains formed pellets, and the optimal conditions for bio-capsule formation were $28^{\circ}C$ and 120 rpm. Lastly, SSF process was performed using a bio-capsule. An ethanol yield of 3.9% was achieved by using the Aspergillus sp. BCNU 6200 bio-capsule (Aspergillus sp. BCNU 6200 + S. cerevisiae IFO-M-07) at $30^{\circ}C$ with shaking at 120 rpm during the 10 days of incubation. The results provide useful information on the application of a bio-capsule in bioethanol production under the SSF process.

• Microbiology and Biotechnology Letters
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• v.8 no.2
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• pp.93-102
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• 1980

Fungi which were capable of producing gluconic acid were isolated from soil and tree leave samples, which had been collected in Seoul ana its vicinity. Among the 19 strains isolated, a strain named arbitrarily KUF-O4 was selected as a test strain chiefty because of its efficiency in gluconic acid production. The strain was identified as an Aspergillus sp. through its morphological properties. Optimum conditions for the gluconic acid production of KUF-O4 were investigated. The results obtained are as follows. 1. An incubation period of at least 30 hours was required for a good yield of gluconic acid. 2. A medium containing 10％ glucose needed at least 3 ％ CaCo$_3$to maintain the optimum pH for the production of gluconic acid during fermentation. 3. As a carbon source, glucose was the most effective one among the carbon sources tested. 4. As a nitrogen source, an ammonium salt was more effective than any other form of nitrogen compounds. 5. As mineral source, a small amount of both KH$_2$PO$_4$and MgSO$_4$was found to be necessary to increase the efficiency of the gluconic acid production.

• Journal of Life Science
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• v.20 no.2
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• pp.269-274
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• 2010

An antifungal antibiotic-producing strain, BCNU 2003, was isolated from forest soil in Korea. The morphological and physiological characters, and 16S rRNA sequences analysis of strain BCNU 2003 identified this strain as Bacillus genus. The Bacillus sp. BCNU 2003 showed strong antifungal activities against Aspergillus niger, Trichophyton mentagrophytes and Trichophyton rubrum with inhibition ranging from 62.05 to 63.49% by using dual culture technique. Bacillus sp. BCNU 2003 produced a maximum level of antifungal substances under aerobic incubation at 28oC and pH 6.5-7.2 for 6 days in LB broth. Ethyl acetate extract of the cultured broth showed strong antifungal activity and a broad antifungal spectrum against various pathogenic fungi. The minimum inhibitory concentration (MIC) values for its active extracts ranged between 0.0625 mg/ml and 1 mg/ml. In addition, Bacillus sp. BCNU 2003 was determined to have the ability to produce enzymes such as amylase, protease, gelatinase and catalase.

• ALGAE
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• v.31 no.4
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• pp.351-362
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• 2016

We investigated a possible reduction in $CO_2$ uptake rate by phototrophic red tide dinoflagellates arising from mixotrophy. We measured the daily ingestion rates of Prorocentrum minimum by Prorocentrum micans over 5 days in 10 L experimental bottles, and the uptake rates of total dissolved inorganic carbon ($C_T$) by a mixture of P. micans and P. minimum(mixotrophic growth), and for the predator P. micans (phototrophic growth; control) and prey P. minimum (phototrophic growth; control) alone. To account for the effect of pH on the phototrophic growth rates of P. micans and P. minimum, measurements of $C_T$ and pH in the predator and prey control bottles were continued until the pH reached the same level (pH 9.5) as that in the experimental bottles on the final day of incubation. The measured total $C_T$ uptake rate by the mixture of P. micans and P. minimum changed from 123 to $161{\mu}mol\;C_T\;kg^{-1}\;d^{-1}$ over the course of the experiment, and was lower than the $C_T$ uptake rates shown by P. micans and P. minimum in the predator and prey control bottles, respectively, which changed from 132 to $17{\mu}mol\;C_T\;kg^{-1}\;d^{-1}$ over the course of the experiment. The reduction in total $C_T$ uptake rate arising from the mixotrophy of P. micans was 7-31% of the daily $C_T$ uptake rate seen during photosynthesis. The results suggest that red tide dinoflagellates take up less $C_T$ during mixotrophy.

• Journal of Mushroom
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• v.10 no.1
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• pp.3-8
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• 2012

In this study, to produce Flammulina velutipes mushroom liquid spawn efficiently and effectively the effects of explosive aeration (supplying air with tiny bubbles) of the liquid culture medium on carbon dioxide concentration and residual sugar content in the medium on carbon dioxide concentration and residual sugar contentin the medium were measured. Carbon dioxide concentrations were measured at the outlet of the incubator. On the third day the explosive aeration greatly increased mycelial growth of the liquid spawn, and carbon dioxide concentration also greatly increased but decreased after 5 days. Free sugar contents in the liquid culture consistantly decreased up to 7 days and thereafter was not detected. The weight of the mycelia were maintained similar levels after 3 days. Total nitrogen content in the liquid medium constantly decreased during the 11days of explosive aeration. The content of free sugars in 7 days of culture was the lowest level, thus the inoculum incubated for 6~7 days was thought to be the most effective. Carbon dioxide concentration measurement at the outlet of the container during the liquid spawn incubation required low cost but was efficient to estimate the degree of mycelial growth to be used as a simple indicator.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.9
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• pp.1273-1278
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• 2016

In this study, we optimized conditions for extraction of eleutherosides B and E from stem powder of Acanthopanax. To enhance eleutheroside B and E yields, Acanthopanax sessiliflorus Seeman and Eleutherococcus senticosus Maxim were pre-incubated with the following enzymes: Celluclast, Viscozyme L, Lactozyme, Lecitase, and Novozyme 33095. Treatment with Novozyme 33095, a commercial pectinase, for 3 h resulted in the highest yields of eleutherosides B and E from A. sessiliflorus and E. senticosus. Compared with extraction at $121^{\circ}C$ for 120 min, at $121^{\circ}C$ for 15 min after Novozyme 33095 pre-incubation increased eleutheroside B and E yields from A. sessiliflorus by 7% and 17%, respectively. In the case of E. senticosus, eleutheroside B and E yields increased by 25% and 29%, respectively.