• The Korean Journal of Mycology
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• v.25 no.4 s.83
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• pp.311-319
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• 1997

These studies were carried out to develop an artificial cultivation method. The diameter and thickness of pileus ranged $1.5{\sim}7.0\;cm$ and $0.8{\sim}3.0\;cm$, respectively. The diameters of stipe were $1.2{\sim}2.5\;cm$ and the lengthes were $4.5{\sim}9.0\;cm$. The spore fingerprint was white. The sizes were spore $10.8{\sim}12.2{\times}4.35{\sim}5.65\;{\mu}m$, basidia $50.0{\sim}59.2{\times}7.4{\sim}7.8\;{\mu}m$, nalsistidia $21.75{\sim}28.7{\times}4.8{\sim}6.1\;{\mu}m$, pileus hymenium cell $50.6{\sim}66.0{\times}4.4{\sim}6.7\;{\mu}m$, and stipe hymenium cell $28.6{\sim}33.0{\times}5.5{\sim}6.6\;{\mu}m$. The thirty percent mixture of rice and wheat bran into sawdust gives the high density of mycelia and the good development of fruiting structure. The optimum water contents of sawdust substrates were $60{\sim}65%$ in which condition the mycelium grows well and gives high density. In PP bottle cultivation, the first fruiting period was $6{\sim}8$ days earlier in nonscratching samples than scratching ones, but the quantity of fruiting body was higher in scratching samples than nonscratching ones. In the case of PP bag cultivation, the first fruiting was 10 days faster, and the quantity of fruiting bodies was 30% higher in samples with 30% wheat bran than those with rice bran. The fleshiness of stipe was $2{\sim}3$ times harder than that of pileus.

• Journal of Mushroom
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• v.3 no.4
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• pp.140-144
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• 2005

This study was conducted to verify the cause of suppression symptom in mycelial growth during summer and to be able to establish a countermeasure. Cultivation of Flammulina velutipes was experimented with varying elapsed time of 0, 3, 6, 9 hours after mixing the sawdust media and two kinds of water temperature (normal water, $24^{\circ}C$ and cold water, $6^{\circ}C$) for mixing sawdust media. There were trends of increased media temperature from $24^{\circ}C$ to $31^{\circ}C$ and decreased pH from 6.5 to 5.2~5.6 with varying elapsed time from mixing the media to sterilization. Bacterial density also increased with bacterial density in Medium $24^{\circ}C$ being 1.9~4.1 times higher than that in Medium 6. Growth of F. velutipes was delayed with dual culture of bacteria isolated from sawdust media. Total nitrogen content of sawdust media was lowered by elapsed time from mixing the media to sterilization. The use of normal water($24^{\circ}C$) delayed mushroom growth by 1~2 days compared with that of cold water($6^{\circ}C$). Furthermore, mycelial growth of F. velutipes in the bottle cultivation ceased 9 hours after mixing the media. Primordia formation of F. velutipes was delayed by 1~3 days by elapsed time after mixing sawdust media, while fruit-body yield decreased by 7~12% 6 hours after mixing the media. Fruit-body yield was more stabilized with the use of cold water($6^{\circ}C$) than with that of normal water($24^{\circ}C$). Results showed that it is effective to use cold water as low as $6^{\circ}C$ in mixing media for F. velutipes cultivation, especially during summer when environmental temperature is high, high pressure sterilization after bottling work can prevent bacterial propagation in the media and can stabilize media ingredient.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.6
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• pp.1230-1236
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• 2012

To verify that the electrokinetic remediation is effective for decreasing salinity of fields of the plastic-film house, field tests for physical property, chemical property, and crop productivity of soils have been conducted. The abridged result of those tests is as follows. In the EK treatment, the electrokinetic remediation has been treated at the constant voltage (about 0.8 V $cm^{-1}$) for fields of the farm household. At this time, an alternating current (AC) 220 V of the farm household was transformed a direct current. The HSCI (High Silicon Cast Iron) that the length of the stick for a cation is 20cm, and the Fe Plate for an anion have been spread out on the ground. As the PVC pipe that is 10 cm in diameter was laid in the bottom of soils, cations descend on the cathode were discharged together. For soil physical properties according to the EK treatment, the destruction effect of soil aggregate was large, and the infiltration rate of water was increased. However, variations of bulk density and porosity were not considerable. Meanwhile, in chemical properties of soils, principal ions of such as EC, $NO_3{^-}$-N, $K^+$, and $Na^+$ were better rapidly reduced in the EK treated control plot than in the untreated control plot. And properties such as pH, $P_2O_5$ and $Ca^{2+}$ had a small impact on the EK. For cropping season of crop cultivation according to the EK treatment, decreasing rates of chemical properties of soils were as follows; $NO_3{^-}$-N 78.3% > $K^+$ 72.3% > EC 71.6% $$\geq_-$$ $Na^+$ 71.5% > $Mg^{2+}$ 36.8%. As results of comparing the experimental plot that EK was treated before crop cultivation with it that EK was treated during crop cultivation, the decreasing effect of chemical properties was higher in the case that EK was treated during crop cultivation. After the EK treatment, treatment effects were distinct for $NO_3{^-}$-N and EC that a decrease of nutrients is clear. However, because the lasting effect of decreasing salinity were not distinct for the single EK treatment, fertilization for soil testing was desirable carrying on testing for chemical properties of soils after EK treatments more than two times. In the growth of cabbages according to the EK treatment, the rate of yield increase was 225.5% for the primary treatment, 181.0% for the secondary treatment, and 124.2% for third treatment compared with the untreated control plot. The yield was increased by a factor of 130.0% for the hot pepper at the primary treatment (Apr. 2011), 248.1% for the lettuce at the secondary treatment (Nov.2011), and 125.4% for the young radish at the third treatment (Jul. 2012). In conclusion, the effect of yield increase was accepted officially for all announced crops.

• Food Engineering Progress
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• v.14 no.3
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• pp.208-216
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• 2010

Lab scale experiments were conducted in order to assess the applicability of $CaCO_{3}$-alginate beads as neutralizer for the high cell density cultivation and prepare the direct vat inoculation cultures of isolated sourdough lactic acid bacteria. With increasing the amount of bead and decreasing the diameter of bead in acidic solution, the neutralizing effect of $CaCO_{3}$-alginate bead became higher. In batch process with $CaCO_{3}$-alginate beads, Lactobacillus amylovorus DU-21 isolated from sourdough showed the highest viable cell counts and optical density in MRS broth. The values of viable cell counts and optical density were 9.996 log CFU/mL and 3.97, respectively. Experiments on the conditions which increase viability during lyophilization were carried out and the following results were obtained; 15% glycerol revealed the high cryoprotective effect on the concentrated cultures during lyophilization among the two cryoprotective agents. Consequently, $CaCO_{3}$-alginate beads and 15% glycerol were found to be useful not only to cultivate Lactobacillus amylovorus DU-21 but also to preserve strain.

• KSBB Journal
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• v.13 no.6
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• pp.681-686
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• 1998

Effect of glucose feeding strategy and initial concentration of glucose on Serratia marcescens ATCC 27117 in high cell density fed-batch fermentation was investigated. The final biomasses in batch, constant feeding, constant and exponentially feeding strategy at glucose starvation condition in fed-batch were 1.40, 5,07, 6,93 and 7.60 g/L at 40, 41, 24 and 40 hrs, respectively. Productivities of biomass were 0.035, 0.124, 0.289 and 0.190 g/L$.$h, respectively. As a result, constant feeding strategy at starvation condition was 1.5∼8.6 times higher than other strategies. The relationship between dissolved oxygen and glucose feeding times was good identified in exponential feeding strategy and constant feeding strategy at starvation condition. And high cell density cultivation was obtained when minimal media was used.

• Korean Journal of Soil Science and Fertilizer
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• v.37 no.3
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• pp.165-170
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• 2004

This study was conducted to investigate the effect of different surface soil management practices on soil microflora in volcanic ash soils of citrus orchard. Soil samples were collected from citrus orchards of clean cultivation, grass sod, and grass mulch system in May and September 1997. Soil chemical properties, populations of various microorganisms, enzyme activities, microbial biomass C were analyzed. Average soil pH were 4.7, and average nitrogen and organic matter contents were 6 and $140.2g\;kg^{-1}$, respectively. Aerobic bacteria were distributed at $26,2-47.3{\times}10^6cfu\;g^{-1}$ level. Among the aerobic bacteria Pseudomonas spp., Rhizobium spp., and thermophilic Bacillus spp. were dominant in most of the investigated orchard soils. Density of actinomycetes were low at $1.8-84.6{\times}10^5cfu\;g^{-1}$ level. Fungi were distributed at $26.4-182.1{\times}10^5cfu\;g^{-1}$ level and the density was higher in grass mulch and sward sites. In september, phosphomonoesterase activity was high at $239.6{\mu}g\;PNP\;g\;soil^{-1}\;h^{-1}$ in clean cultivated citrus orchards. Soil cellulase activity were higher at $602.6{\mu}g\;GE\;g\;soil^{-1}$\;24\;h^{-1}$ in grass sward cultivation than any other soil management practices. Soil microbial biomass C was higher in grass mulch cultivated orchards.

• Journal of Microbiology and Biotechnology
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• v.31 no.10
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• pp.1430-1437
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• 2021

Cronobacter sakazakii is an opportunistic pathogenic bacterium found in powdered infant formula and is fatal to neonates. Antibiotic resistance has emerged owing to overuse of antibiotics. Therefore, demand for high-yield bacteriophages as an alternative to antibiotics has increased. Accordingly, we developed a modified mass-production method for bacteriophages by introducing a two-stage self-cycling (TSSC) process, which yielded high-concentration bacteriophage solutions by replenishing the nutritional medium at the beginning of each process, without additional challenge. pH of the culture medium was monitored in real-time during C. sakazakii growth and bacteriophage CS01 propagation, and the changes in various parameters were assessed. The pH of the culture medium dropped to 5.8 when the host bacteria reached the early log phase (OD₅₄₀ = 0.3). After challenge, it decreased to 4.65 and then recovered to 4.94; therefore, we set the optimum pH to challenge the phage at 5.8 and that to harvest the phage at 4.94. We then compared phage production during the TSSC process in jar-type bioreactors and the batch culture process in shaker flasks. In the same volume of LB medium, the concentration of the phage titer solution obtained with the TSSC process was 24 times higher than that obtained with the batch culture process. Moreover, we stably obtained high concentrations of bacteriophage solutions for three cycles with the TSSC process. Overall, this modified TSSC process could simplify large-scale production of bacteriophage CS01 and reduce the unit cost of phage titer solution. These results could contribute to curing infants infected with antibiotic-resistant C. sakazakii.

• Preventive Nutrition and Food Science
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• v.2 no.4
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• pp.354-359
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• 1997

Optimal cultivation conditions for the production of phenylalanine ammonia-lyase(PAL) from Rhodotorula aurantiaca K-505 were selected, and the kinetic parameters of the produced PAL were determined. The most suitable carbon and nitrogen sources were glucose and tryptone, respectively. The strain expressed PAL constituttively when using the optimized semi-complex media. High cell density culture could be critical for maximal production of PAl since the PAL ynthesis was growth associated. maximum PAL activity was observed at initial pH 6.0. although the ll growth was not markedly affected by temperature between 22 and 28$^{\circ}C$, the cells yielded the maximum PAL activity when cultivated at 22$^{\circ}C$. The maximum activity for deamination of L-phenylalnine to trans-cinnamic acid was observed around pH 8.8. The PAL activity gave the maximum at 45$^{\circ}C$, and greatly decreased at higher than 5$0^{\circ}C$. Activation energy({TEX}$E_{a}${/TEX}) calculated from Arrhenius equation was 6.28 kcal/mol in the range of 22$^{\circ}C$ to 4$0^{\circ}C$. A oolf plot showed that the enzyme reaction follows Michaelis-Menten equation, whose {TEX}$K_{M}${/TEX} and {TEX}$V_{max}${/TEX} values were 4.65$\times${TEX}$10^{-3}${/TEX} M and 0.89$\mu$ mol/mg-min respectively.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.554-556
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• 2003

This research was focused on expression, purification and characterization of ubiquitin-specific protease 1 (UBP1) expressed in recombinant Escherichia coli. Various systems were constructed by fusing polycationic fusion tails or fusion partners to the C- or N-terminus of the product protein. In particular, UBP1 containing 6 histidine residues at the N-terminal end showed best results in terms of expression level and purification efficiency. The N-terminal $6{\times}His-tagged$ UBP1 was overproduced in recombinant E. coli using high cell density cultivation technology and purified using immobilized metal affinity chromatography. The molecular weight of UBP1 was found to be 83,500 daltons. The optimum temperature and pH for the enzyme reaction when ubiquitin-human growth hormone (hGH) was used as a substrate were $40^{\circ}C$ and pH 8.0, respectively.

• Journal of Microbiology and Biotechnology
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• v.10 no.4
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• pp.551-553
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• 2000

A strong constitutive $P_{JH}$ promoter from Bacillus was applied to overexpress the endoxylanase gene in B. subtilis. The expression plasmid, pHJKJ4, was designed to contain the $P_{JH}$ promoter and endoxylanase promoter ($P_B$), and introduced into B. subtilis DB104. Through batch fermentation of the trasformant cell on a maltose medium, endoxylanase was produced in a growth-associated manner as the predominant protein. The total activity reached about 600 unit/ml at the end of the cultivation, which corresponded to 698 mg endoxylanase protein/l with a specific activity of 860 unit/mg protein. It was also found that the segregational plasmid instability was less than 30% and most of the endoxylanase activity was detected in the culture medium. This result suggests that the secretory production of endoxylanase can be significantly enhanced with the use of the $P_{JH}$ promoter and high-cell density culture techniques, quantitatively as well as qualitatively.