• 생명과학회지
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• 제6권2호
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• pp.94-103
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• 1996

To understand effect of inoculum size, cell density, sucrose concentration and concentrations of MS basal on suspension culture and protoplast isolation of wild viola(Viola patrinii DC.) callus from petiole segments this experiment was conducted. In the lot of 30 mesh inoculum size, two observations were; One was that a considerable increase in the fresh and dry weight of callus was determined. Another was that the callus mass was relatively compact compared with others. A recommendable cell density was 0.4g for 20ml culture medium and the higher sucrose concentration, the higher fresh and dry weight were obtained. The dilution of MS basal salt was differently affected on fresh and dry weight; the highest fresh weight was found in 1x MS salt, while the higest dry weight was in 1/3x dilution.The addition of casein hydrolysate(3g/L) was more effective to increase of both fresh and dry weight. THe contents of protein was great in the inoculum lots with larger inoculum sizeand higher concentration of MS basal salts contenting 3g/L of casein hydrolysate and higher sucrose compared with others. The greatest protoplasts were isolated from the lot of 10 mesh size treated with 1%pectinase SE-150 and 2% cellulase YC. In general, for optimal protoplast isolation the followingconditions were recommended; 1) Use of smaller cell size cultured for 2-5 weeks, and 2) more than 5 hours incubation using the combined mixture of the enzymes with proper concentrations.

• 생명과학회지
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• 제14권2호
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• pp.368-373
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• 2004

광범위 제초제인 Bast $a^{(R)}$에 대해 저항성을 가지는 형질전환체 벼를 개발하였다. Bar유전자를 함유하고 있는 플라스미드 pCaMV35S::Bar를 embryogenic 현탁 배양체로부터 분리한 벼의 원형질체에 도입하였다. Phosphinotricin에 대해 저항성을 가지는 형질전환체 식물체들이 재분화되었고, 이들을 15 mg/l phosphinotricin이 함유한 배지에서 다시 선별하였다. 형질전환체 벼에서 bar유전자의 삽입과 발현을 Southern과 Northern blot분석으로 확인하였고, 또한 $R_1$ 형질전환 식물체들을 PAT 활성 assay로 재차 유전자 발현을 확인하였다. Bar 유전자는 다음 세대인 $R_1$ 식물체에서 3:1 멘델 유전 양상을 나타내었고, 형질전환체 $R_1$과 $R_2$ 식물체들은 fieild에서 살포되는 제초제 양만큼 Bast $a^{(R)}$ 를 살포했을 때 제초제 저항성을 나타내었다..

• Preventive Nutrition and Food Science
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• 제21권3호
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• pp.271-280
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• 2016

The effects of $CO_2$ injection and barrel temperatures on the physiochemical and antioxidant properties of extruded cereals (sorghum, barley, oats, and millet) were studied. Extrusion was carried out using a twin-screw extruder at different barrel temperatures (80, 110, and $140^{\circ}C$), $CO_2$ injection (0 and 500 mL/min), screw speed of 200 rpm, and moisture content of 25%. Extrusion significantly increased the total flavonoid content (TFC) of extruded oats, and ${\beta}$-glucan and protein digestibility (PD) of extruded barley and oats. In contrast, there were significant reductions in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, PD of extruded sorghum and millet, as well as resistant starch (RS) of extruded sorghum and barley, and total phenolic content (TPC) of all extrudates, except extruded millet. At a barrel temperature of $140^{\circ}C$, TPC in extruded barley was significantly increased, and there was also an increase in DPPH and PD in extruded millet with or without $CO_2$ injection. In contrast, at a barrel temperature of $140^{\circ}C$, the TPC of extruded sorghum decreased, TFC of extruded oats decreased, and at a barrel temperature of $110^{\circ}C$, PD of extruded sorghum without $CO_2$ decreased. Some physical properties [expansion ratio (ER), specific length, piece density, color, and water absorption index] of the extrudates were significantly affected by the increase in barrel temperature. The $CO_2$ injection significantly affected some physical properties (ER, specific length, piece density, water solubility index, and water absorption index), TPC, DPPH, ${\beta}$-glucan, and PD. In conclusion, extruded barley and millet had higher potential for making value added cereal-based foods than the other cereals.

• Mycobiology
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• 제30권1호
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• pp.31-36
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• 2002

Soil bacteria were screened for the ability to control cucumber anthracnose caused by Colletotrichum orbiculare through induced systemic resistance(ISR). Sixty-four bacterial strains having in vitro antifungal activity were used for selecting ISR-inducing strains in cucumber. Cucumber seeds(cv. Baeknokdadagi) were sown in potting mixtures incorporated with the soil bacteria, at a rate of ca. $10^8$ cells per gram of the mixture. Two week-old plants were then transplanted into the steam-sterilized soil. Three leaf-stage plants were inoculated with a conidial suspension($5{\times}10^5$ conidia/ml) of C. orbiculare. Diseased leaf area(%) and number of lesions per $cm^2$ leaf were evaluated on third leaves of the plants, $5{\sim}6$ days after inoculation. Among 64 strains tested, nine strains, GC-B19, GC-B35, GK-B18, MM-B22, PK-B14, RC-B41, RC-B64, RC-B65, and RC-B77 significantly(P=0.05) reduced anthracnose disease compared to the untreated control. In contrast, some bacterial strains promoted susceptibility of cucumber to the disease. From the repeated experiments using the nine bacterial strains, GC-B19, MM-B22, PK-B14, and RC-B65 significantly(P=0.05) reduced both diseased leaf area(%) and number of lesions per $cm^2$ leaf in at lease one experiment. These strains with control efficacy of $37{\sim}80%$ were determined to be effective ISR-inducing strains.

• 한국물환경학회지
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• 제33권5호
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• pp.531-537
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• 2017

The biologic Oxygen Demand (BOD) is a reliable and generally accepted indicator of water pollution by organic pollutants. Accordingly, estimation of BOD export from paddies carries important implications fwith regard to water management in rural areas. In this study, hydrology and BOD concentration were monitored during the period 2008 through 2012, in an effort to understand the characteristics of BOD export from paddy fields. The findings demonstrated that BOD load by rainfall above 50 mm. occupied about 50 % of total load, whereas the load by less than ten mm. rainfall occupied about 29 % of the total load during periods of stormy activity. It therefore seems that it could be possible to reduce the BOD load up to 29 % during storm periods, when drainage control conducted for rainfall less than ten mm.(an amount which is relatively easy to manage). The documented mean loads of storm and non-storm were $17.1kg\;ha^{-1}\;yr^{-1}$ and $11.2kg\;ha^{-1}\;yr^{-1}$, respectively. The BOD load during the significant rainfall period was similar to the renewed unit load by NIER (2014). However, there were substantial differences between unit load and actual load when the non-storm load was incorporated into the BOD load estimation from paddy fields. In view of the foregoing, it is felt that, the non-storm load needs to be further considered and managed for the successful implementation of Total Maximum Daily Load (TMDL) program.

• 한국축산식품학회지
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• 제34권3호
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• pp.333-338
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• 2014

The objective of this study was to investigate the effects of two different ripening durations, with, or without adding rosemary powder, on Lomo embuchado (LEO) sausage quality. All LEOs were ripened for two different durations, 45 or 60 d, with, or without the addition of rosemary powder, as follows: 1) LEO ripened for 45 d (LER45), 2) LEO ripened for 60 d (LER60), 3) rosemary LEO ripened for 45 d (RLE45), and 4) rosemary LEO ripened for 60 d (RLE60). Significant differences were observed in both moisture and ash content, with higher moisture and less ash content in LER45 (p<0.05). No trend was shown in the crude protein content of the four different treatments, but significantly low protein content was shown only in RLE45 (p<0.05). Ripening for 45 d improved the lightness, yellowness, and water activity of LEOs (p<0.05). However, ripening duration together with rosemary powder addition had no significant effects on redness (p>0.05). The LER45 generated significantly improved chewiness, gumminess, and hardness, as compared to both LER60 and RLE60 (p<0.05). In conclusion, the results suggest that ripening for 45 d seems to enhance LEO quality, but that rosemary powder addition may not be required to develop good LEO quality.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.248-254
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• 2016

A soil metagenome contains the genomes of all microbes included in a soil sample, including those that cannot be cultured. In this study, soil metagenome libraries were searched for microbial genes exhibiting lipolytic activity and those involved in potential lipid metabolism that could yield valuable products in microorganisms. One of the subclones derived from the original fosmid clone, pELP120, was selected for further analysis. A subclone spanning a 3.3 kb DNA fragment was found to encode for lipase/esterase and contained an additional partial open reading frame encoding a wax ester synthase (WES) motif. Consequently, both pELP120 and the full length of the gene potentially encoding WES were sequenced. To determine if the wes gene encoded a functioning WES protein that produced wax esters, gas chromatography-mass spectroscopy was conducted using ethyl acetate extract from an Escherichia coli strain that expressed the wes gene and was grown with hexadecanol. The ethyl acetate extract from this E. coli strain did indeed produce wax ester compounds of various carbon-chain lengths. DNA sequence analysis of the full-length gene revealed that the gene cluster may be derived from a member of Proteobacteria, whereas the clone does not contain any clear phylogenetic markers. These results suggest that the wes gene discovered in this study encodes a functional protein in E. coli and produces wax esters through a heterologous expression system.

• Journal of Microbiology and Biotechnology
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• 제21권9호
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• pp.947-953
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• 2011

A fungal strain, Aspergillus terreus strain GA2, isolated from an agricultural field cultivating sweet sorghum, produced feruloyl esterase using maize bran. In order to obtain maximum yields of feruloyl esterase, the solid state fermentation (SSF) conditions for enzyme production were standardized. Effective feruloyl esterase production was observed with maize bran as substrate followed by wheat bran, coconut husk, and rice husk among the tested agro-waste crop residues. Optimum particle size of 0.71-0.3 mm and moisture content of 80% favored enzyme production. Moreover, optimum feruloyl esterase production was observed at pH 6.0 and a temperature of $30^{\circ}C$. Supplementation of potato starch (0.6%) as the carbon source and casein (1%) as the nitrogen source favored enzyme production. Furthermore, the culture produced the enzyme after 7 days of incubation when the C:N ratio was 5. Optimization of the SSF conditions revealed that maximum enzyme activity (1,162 U/gds) was observed after 7 days in a production medium of 80% moisture content and pH 6.0 containing 16 g maize bran [25% (w/v)] of particle size of 0.71-0.3 mm, 0.6% potato starch, 3.0% casein, and 64 ml of formulated basal salt solution. Overall, the enzyme production was enhanced by 3.2-fold as compared with un-optimized conditions.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.316-325
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• 2012

Xylanase has been used extensively in the industrial and agricultural fields. However, the low-yield production of xylanase from native species cannot meet the increasing demand of the market. Therefore, improving the heterologous expression of xylanase through basic gene optimization may help to overcome the shortage. In this study, we synthesized a high-GC-content native sequence of the thermostable xylanase gene xynB from Streptomyces olivaceoviridis A1 and, also designed a slightly AT-biased sequence with codons completely optimized to be favorable to Pichia pastoris. The comparison of the sequences' expression efficiencies in P. pastoris X33 was determined through the detection of single-copy-number integrants, which were quantified using qPCR. Surprisingly, the high GC content did not appear to be detrimental to the heterologous expression of xynB in yeast, whereas the optimized sequence, with its extremely skewed codon usage, exhibited more abundant accumulation of synthesized recombinant proteins in the yeast cell, but an approximately 30% reduction of the secretion level, deduced from the enzymatic activity assay. In this study, we developed a more accurate method for comparing the expression levels of individual yeast transformants. Moreover, our results provide a practical example for further investigation of what constitutes a rational design strategy for a heterologously expressed and secreted protein.

• Journal of Microbiology and Biotechnology
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• 제23권2호
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• pp.161-166
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• 2013

At present, acetylcholinesterase (AChE) inhibitors are the first group of drugs to treat mild to moderate Alzheimer's disease (AD). Although beneficial in improving cognitive and behavioral symptoms, the effectiveness of AChE inhibitors has been questioned since they do not delay or prevent neurodegeneration in AD patients. Therefore, in the present study, in order to develop new and effective anti-AD agents from lichen products, both the AChE inhibitory and the neuroprotective effects were evaluated. The AChE inhibitory assay was performed based on Ellman's reaction, and the neuroprotective effect was evaluated by using the MTT method on injured PC12 cells. One AChE inhibitor ($IC_{50}$ = 27.1 ${\mu}g/ml$) was isolated by means of bioactivity-guided isolation from the extract of lichen-forming fungus Cladonia macilenta, which showed the most potent AChE inhibitory activity in previous screening experiment. It was then identified as biruloquinone by MS, and $^1H$- and $^{13}C$-NMR analyses. The inhibitory kinetic assay suggested that biruloquinone is a mixed-II inhibitor on AChE. Meanwhile, biruloquinone improved the viability of the $H_2O_2$- and ${\beta}$-amyloid-injured PC12 cells at 1 to 25 ${\mu}g/ml$. The protective effects are proposed to be related to the potent antioxidant activities of biruloquinone. These results imply that biruloquinone has the potential to be developed as a multifunctional anti- AD agent.