• Journal of Life Science
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• v.14 no.5
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• pp.840-846
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• 2004

A study for expression of Korean Mistletoe (KM) lectin gene (A,B) in Saccharomyces cerevisiae was done using transforming system of yeast. In order to overexpress the genes efficiently in yeast, two lectin genes (A,B) were re-cloned and modified including Kozak translation initiation sequence using PCR amplification. The constructed plasmids containing modified lectin A and B genes were transformed to S. cerevisea INVSc (MAT G, his3 $\Delta$1, leu2, trpl-289, ura3-52). The transformed cells were identified by DNA sequencing with ABI3700 system and induced with 2% of galactose for recombinant KM lectin (rKM lectin) protein. The rKM lectin A and B proteins were determinated about 29kDa size of protein by SOS-P AGE and western blotting analysis. The expressed recombinant lectin was determinated 1.24∼1.75 $\mu\textrm{g}$ per 1 mg of cytosolic soluble protein by sandwich ELISA method. Moreover the lectin genes were expressed as maximum level at 36 h after galactose induction and lectin A gene was were repressed after 48 h.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.163-167
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• 2012

Since D-xylose is not fermentable in Saccharomyces cerevisiae, its conversion to D-xylulose is required for its application in biotechnological industries using S. cerevisiae. In order to convert D-xylose to D-xylulose by way of an enzyme immobilized system, D-xylose isomerase (XI) of Escherichia coli was fused with 10-arginine tag (R10) at its C-terminus for the simple purification and immobilization process using a cation exchanger. The fusion protein XIR10 was overexpressed in recombinant E. coli and purified to a high purity by a single step of cation exchange chromatography. The purified XIR10 was immobilized to a cation exchanger via the electrostatic interaction with the C-terminal 10-arginine tag. Both the free and immobilized XIR10 exhibited similar XI activities at various pH values and temperatures, indicating that the immobilization to the cation exchanger has a small effect on the enzymatic function of XIR10. Under optimized conditions for the immobilized XIR10, D-xylose was isomerized to D-xylulose with a conversion yield of 25%. Therefore, the results of this study clearly demonstrate that the electrostatic immobilization of XIR10 via the interaction between the 10-arginine tag and a cation exchanger is an applicable form of the conversion of D-xylose to D-xylulose.

• Journal of Life Science
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• v.27 no.2
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• pp.172-179
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• 2017

The fermentation of Panax ginseng can yield many compounds from ginsenosides that have a wide variety of biological functions. Lactic acid bacteria (LAB) strains are capable of converting ginsenosides. The purposes of this study were to: (1) characterize Enterococcus faecium SA5, an isolated LAB from Mongolian mare milk, (2) identify the existence of extracellular ${\beta}$-glucosidase activity in the milk, and (3) ascertain if the ${\beta}$-glucosidase has the capacity of converting ginsenoside in Korean ginseng. The results revealed that E. faecium SA5 was acid-resistant, bile salt-resistant, and has antibiotic activities against 4 pathogenic microorganisms (Salmonella typhimurium KCTC 3216, Listeria monocytogenes KCTC 3710, Bacillus cereus KCTC 1012, Staphylococcus aureus KCTC 1621). In addition, E. faecium SA5 had tolerance against some antibiotics such as colistin, gentamycin and neomycin. It was also found that E. faecium SA5 possessed bile salt hydrolase activity, which could lower blood cholesterol level. When incubated in 10% (w/v) skim milk as a yogurt starter, E. faecium SA5 caused to decrease pH of the medium as well as increase in viable cell counts. Using TLC and HPLC analysis on the samples incubated in MRS broth, our study confirmed that E. faecium SA5 can produce ${\beta}$-glucosidase, which was capable of converting ginsenoside $Rb_1$ into new ginsenosides $Rg_3-s$ and $Rg_3-r$. It was concluded that E. faecium SA5 possessed a potential of probiotic activity, which could be applied to yogurt manufacture as well as ginsenoside conversion in ginseng.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.155-160
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• 1986

$\alpha$-Amylase gene of B. amyloliquefaciens was cloned to E. coli-yeast shuttle vector YEp-13 and expressed in E. coli. Chromosomal DNA of B. amyloliquefaciens was partially digested with Sau3Al and YEp13 plasmid was cleaved with BamH1. The hybrid plasmid, pHA28, was constructed by shotgun method and transformed to E. coli C600 and HB101. The amount of $\alpha$-amylase produced by transformants of E. coli was about 20% to 30% of that produced by B. amyloli-quefaciens. About 65% of $\alpha$-amylase produced by transformant was secreted into periplasm and the others were located in cytoplasm. $\alpha$-Amylase production was maximal when transformants were cultivated for 15hr to 20hr. As the result of agarose gel electrophoresis, pHA28 plasmid was found to be various in its size. This result suggested that pHA28 plasmid was segregated.

• Journal of The Korean Society of Grassland and Forage Science
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• v.21 no.3
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• pp.151-156
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• 2001

This study was conducted to obtain the transformed alfalfa (Medicago sativa L.) plants with thermotolerance gene (BcHSP17.6) using Agrobacterium tumefaciens LBA4404 and we confirmed the transformed gene from the regenerated alfalfa plants. The expression vector, pBKH4, harboring BcHSP17.6 gene was used for production of transgenic alfalfa plants. In a process for transformation, the callus of alfalfa was cocultivated with Agrobacterium tumefaciens and transformed calli were selected on kanamycin-containing SH-3-kc medium to regenerate into into the plant. The complete transgenic alfalfa plants were produced by cultivation for about 4 months on several regeneration media, SH-nk-c, SH-l lb-c, SH-sp-c, and SH-IBA. The transgenic alfalfa plants were analyzed by isolation of genomic DNA and PCR/Southem blot.

• Korean Journal of Soil Science and Fertilizer
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• v.17 no.3
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• pp.265-273
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• 1984

To investigate nitrogen supplying effect of some slowrelease N-fertilizers on barley in upland soil sulfurcoated urea(SCU), ureaform and melamine (1,3,5-Triazine-2,4,6-triamine) were treated and compared to urea. In addition, thiourea as a nitrification inhibitor was also tested. Effects of variable soil condition such as moisture content, pH and temperature on nitrogen supplying ability of the fertilizers and on growth of barley were studied through incubation test and pot culture and the obtained results were summarized as follows: The releasing rate of ammonia from urea, SCU, ureaform and melamine were resulted as 27-59%, 25-39%, 9-34% and 0.7-4.3% at maximum conversion rate, respectively. Nitrification rate of the tested fertilizers was higher at pH 6.54 markedly than at pH 4.73. Addition of thourea depressed the formation of $NO_3$ during four weeks of incubation period. Mixed application of ureaform with small amount of urea contributed to nitrogen supply till latter growth stage of barely Basal application of melamine showed lowest nitrogen supplying ability and injurious response on barley growth.

• Journal of the Mineralogical Society of Korea
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• v.30 no.4
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• pp.205-217
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• 2017

Mineral carbonation by indirect method has been studied by serpentinite as cation source. Through the carbonation of $CO_2$ and alkaline earth ions (calcium and magnesium) from serpentinite, the pure carbonates including $MgCO_3$ and $CaCO_3$ were synthesized. The extraction solvent used to extract magnesium (Mg) was ammonium sulfate ($(NH_4)_2SO_4$), and the investigated experimental factors were the concentration of $(NH_4)_2SO_4$, reaction temperature, and ratio of serpentinite to the extraction solvent. From this study, the Mg extraction efficiency of approximately 80 wt% was obtained under the conditions of 2 M $(NH_4)_2SO_4$, $300^{\circ}C$, and a ratio of 5 g of serpentinite/75 mL of extraction solvent. The Mg extraction efficiency was proportional to the concentration and reaction temperature. $NH_3$ produced from the Mg extraction of serpentinite was used as a pH swing agent for carbonation to increase the pH value. About 1.78 M of $NH_3$ as the form of $NH_4{^+}$ was recovered after Mg extraction from serpentinite. And, the main step in Mg extraction process of serpentinite was estimated by geochemical modeling.

• Applied Biological Chemistry
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• v.36 no.6
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• pp.481-487
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• 1993

The production of sorbose from sorbitol in resting cell system of Acetobacter suboxydans was studied. The conversion of sorbose from sorbitol was markedly influenced by several factors such as the substrate concentration, reaction time, temperature, pH, metal ions, growth factors and aeration in the resting cells. Sorbose production rapidly increased in the range of 6 mg/ml cells with the concentration of 5% sorbitol. For production of sorbose from sorbitol, optimal temperature and pH were $30^{\circ}C$ and 6.0. The production of sorbose from sorbitol was activated by 1 mM of $Al^{+3}$ while inhibited by $Ni^{+2}$. The conversion of sorbitol to sorbose was stimulated by the adding of 1 mM p-aminobenzoic acid and nicotinic acid, respectively. During incubation of 1.5 ml of reaction mixture in 50 ml of Erlenmeyer flask, 5% sorbitol was completly converted to sorbose after 20 hours.

• Korean Journal of Microbiology
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• v.36 no.3
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• pp.236-241
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• 2000

A defensin is an inducible antibacterial peptide from a fleshfly and contains 40 residues basic peptide with six cysteines. For the consiruction of recombinant S cerevisiae expressing defensin, the structural gene coding for active defensin was chemically synthesized and fused in fiam to GAP promoter, MFul preprosequence and the GAL7 transcription terminator, generating a recombinant plasnlid pGMD18. S. ce~evisine 2805 Gells were transror~ned to uracil prototroph by the pGMDl8 arid the transformed cells showing antibacterial activity against 111. luteus TAM1056 were selected by growth inhibition zone assay. The optimal culture conditions for the unprovement of the defensin production of a selected tmdonnant were investigated. The optirmzed medium containing 0.4% yeast extract, 2% corn steep liquor, 2.5% glucose and 0.05% $C_2CO_3$, could be determined and the optimum lemperature. and initial pH could be detennnied as $28^{\circ}C$ and pH 3, ~mpectively. The optimized conditioiis revealed the trvofold Increase in the cell growth and the fourfold in the antibaclerial activity. coinpar-ed with tllc Yl'D medium.

• Korean Journal of Soil Science and Fertilizer
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• v.12 no.3
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• pp.117-124
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• 1979

The present work was carried out to study the fate of applied phosphorus labelled with $^{32}P$ and its availability to plants in soils subjected to different management practices. The results can be summarized as follows (Table 3): 1. The applied phosphorus was transformed into different phosphorus compounds in the soils depending upon the management practices and soil characteristics. 2. In the flooded paddy soil (pH 5.8) added P after one week of incubation was transformed into various fractions, the order of abundance being: Al-P> Ca-P$${\sim_\sim}$$Fe-P> Org.-P. After two weeks the order changed to: Fe-P> Al-P> Ca-P> Org.-P. The amounts of the Fe-P and Al-P fractions were found to increase from the second week of incubation whereas a decrease in Ca-P was noticed with the organic-P remaining constant. The amount of available P decreased from the first to the third week of incubation, but increased thereafter. 3. In the volcanic ash soil a major proportion of the applied phosphorus was found in the Fe-P fraction during the whole experimental period. The interconversions of the $^{32}P$ among the different phosphate fractions was not as evident as in the case of flooded rice soil. The recovery of applied P was low and remained constant throughout the incubation period. 4. In the upland soils relatively more of the applied phosphorus was found in the Ca-P fraction as compared with those of the other soils. As in the flooded paddy soil $^{32}P$ in the Ca-P fraction decreased with increasing incubation time, whereas in the Fe-P fraction it increased with time. The recovery of added phosphate as available P followed different patterns for the cultivated and the uncultivated soils. In the cultivated soils lit was relatively high and remained nearly constant during the whole incubation period. In the uncultivated soil on the other hand, it was high at the earlier time of incubation, but decreased with incubation time.