The enzymatic degradation of xylans is the most versatile way to obtain the high value-added functional compounds or the fermentable sugars for renewable energy. The endo-${\beta}$-xylanases are the major enzymes which hydrolyze the internal ${\beta}$-1,4-linkages of xylan backbones to produce the mixtures of xylooligosaccharides including xylobiose and xylotriose. Among them, glucuronoxylanase GH30 can exclusively hydrolyze the internal ${\beta}$-1,4-linkages of xylans decorated with methylglucuronic acid branches. In the present study, two xylanolytic enzyme (PaXN_10 and PaGuXN_30) genes were cloned from Paenibacillus amylolyticus KCTC 3005, and expressed in Escherichia coli, respectively. PaXN_10 (38.7 kDa) belongs to the endo-${\beta}$-xylanases GH10 family, while PaGuXN_30 (58.5 kDa) is a member of glucuronoxylanase GH30. They share the same optimal reaction conditions at $50^{\circ}C$ and pH 7.0. Enzymatic characterization proposed that P. amylolyticus can utilize the hardwood glucuronoarabinoxylans via the cooperative actions of xylanases GH10 and GH30. The extracellular PaGuXN_30 is secreted into the medium and hydrolyzes glucuronoarabinoxylans to release a series of aldouronic acid mixtures with a methylglucuronic acid branch. The resultant products being transported into the microbial cell are successively degraded into the smaller xylooligosaccharides by the intracellular PaXN_10, which will be utilized for the cellular metabolism.
For the environmental friendly soil management on the cultivation of crops in the greenhouse, organic materials, such as the by product-fertilizer derived from livestock manure, rice straw, mushroom media, rice hulls, wood sawdust, and cocopeat, were used as carbon sources adjusting the ratio of carbon to nitrogen to 10, 20, and 30 based on the inorganic soil N. In each C/N ratio of greenhouse soil, watermelon was cultivated in the greenhouse as crop for experiment for the spring and summer of the year and the experimental results were summarized as follows. The concentration of T-C in the organic materials applied were between $289{\sim}429g\;kg^{-1}$, In the C/N ratio of 10, using watermelon as the crop cultivated during the second half of the year in the greenhouse soil, the $NO_3$-N and EC were reduced by 21 to 37%, and 26 to 33%, respectively, except the by product-fertilizer from livestock manure, compared to the soil $NO_3$-N and EC used in the experiment. After the watermelon was cultivated in soils that C/N ratios were controlled as 10, 20, and 30 with wood sawdust adding as carbon sources in the three soils with the different EC values, EC values of the soils were reduced by 33, 42, and 39%, respectively, compared to the soil EC used in the experiment. The weight of watermelon was 10.1-13.4 kg per one unit, and, of the three soils with different EC values. In the soils with three different EC values controlled at C/N ratio of 20, the weight of watermelon was good. The degree of sugar of watermelon were 11.8 to 12.3 Brix, which means that the difference between the treatments was not significant. In conclusion, the C/N ratio of 20 controlled by the proper supply of organic materials according to the representative EC values shown in the greenhouse soils was optimal condition enough to maintain the soil management for the organic culture with the proper nutrient cycling.
Purpose : To Investigate the pathways of radiation induced apoptosls and the effect of cysteamine (${\beta}$-mercaptoethyiamine), as a radioprotector, on it. Materials and Methods : HL-50 ceils were assigned to control, irradiated, and cysteamlne (1 mM, 10mM) pretreated groups. Irradiation was given In a single fraction of 10 Gy (6 MV x-ray) and cysteamine was administered 1 hour before irradiation. The activities of caspase-8 were measured in control and irradiated group to evaluate its relation to the radiation Induced apoptosis. To evaluate the role of cysteamine In radiation Induced apoptosis, the number of viable cells, the expression and activity of caspase-3, and the expression of poly (ADP-ribose) polymerase (PARP) were measured and compared after irradiating the HL-60 celis with cysteamine pretreatment or not. Results : The intraceliular caspase-8 activity, known to be related to the death receptor induced apoptosis, was not affected by irradiation(p>0.05). The number of viable cells began to decrease from 6 hours after irradiation (p>0.05), but the number of viable cells In 1 mM cysteamine pretreated group was not decreased after irradiation and was similar to those in the control group. In caspase-3 analyses, known as apoptosis executioner, its expression was not different but its activity was Increased by irradiation(p>0.05). However, this Increase of activity was suppressed by the pretreatment of 1 mM cysteamine. The cleavage of PARP, thought to be resulted from caspase-3 activation, occurred after irradiation which was attenuated by the pretreatment of 1 mM cysteamine. Conclusion : These results show that radiation induced apoptotic process is somewhat different from death receptor induced one and the pretreatment of 1 mM cysteamine has a tendency to decrease the radiation-induced apoptosis in HL-60 cells.
Kim, Taewan;Lee, Jaemin;Jeong, Gyeong Han;Kim, Tae Hoon
Food Science and Preservation
/
v.23
no.2
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pp.283-289
/
2016
Naturally occurring antioxidants, such as polyphenols are widely found in fruits, vegetables, wines, juices, and other plant-based dietary sources and are divided into several sub classes, including phenylpropanoids, flavonoids, stilbenoids, and lignans. As part of the our ongoing search for bioactive food ingredients, the antioxidant and advanced glycation end products (AGEs) formation inhibitory activities of the methanolic extract of the aerial parts of Cirsium setidens were investigated in vitro bioassay system. The antioxidant properties were evaluated through radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) ($ABTS^+$) radicals. In addition, the activity of C. setidens against diabetes complications was also tested via AGEs formation inhibition assay. The total phenolic contents were determined using a UV-VIS spectrophotometric method. All tested samples showed a dose-dependent radical scavenging and AGEs inhibitory activities. In particular, the n-butanol (BuOH)-soluble portion showed the most potent radical scavenging activities against DPPH and $ABTS^+$ radicals with $IC_{50}$ values of $24.3{\pm}1.7$ and $25.0{\pm}3.3{\mu}g/mL$, respectively. Futhermore, the inhibition of AGEs formation by the n-BuOH-soluble portion ($IC_{50}$ value; $46.0{\pm}1.5{\mu}g/mL$) was higher than that those of the soluble portions for the other solvent. The results showed that C. setidens could be considered as an effective source of natural antioxidants and other ingredients.
Kim, Ji-Ae;Ryoo, Seung-Heui;Yu, Sung-Lan;Lee, Jun-Heon;Seo, Gil-Woong;Kim, Sun-Kyun;Sang, Byung-Chan
Korean Journal of Agricultural Science
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v.29
no.2
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pp.43-52
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2002
This study was performed to provide the basic data for preservation and improvement of genetic resources according to finding genetic construction obtained from analysis of genetic characteristics of $\beta$-casein gene in Korean Native goat and Saanen using the PCR-RFLP. This study confirmed the amplified products of 481bp fragments obtained from the amplification of $\beta$-casein loci by PCR. The $\beta$-casein AB genotype showed 481, 284 and 197bp, and $\beta$-casein BB genotype showed 284 and 197bp fragments in Korean Native goat and Saanen. The frequencies of $\beta$-casein genotype in Korean Native goat were 6.25 and 93.75% for AA and AB and the frequencies of $\beta$-casein genotype in Saanen were 57.14 and 42.86% for AA and AB types. The frequencies of $\beta$-casein A and B alleles were 0.031 and 0.969 in Korean Native goat and the frequencies of $\beta$-casein A and B alleles are 0.286 and 0.714 in Saanen, respectively. The nucleotide sequence of $\beta$-casein gene of Korean Native goat was 97.71% higher homology with 11 nucleotide sequences difference of that of goat reported in GeneBank (M90556). Therefore, this study of molecular genetic characteristics by the analysis of genetic polymorphism and sequencing for $\beta$-casein gene should be used as basic and applying data for preservation and improvement of genetic resources in Korean Native goat breeding.
Kang, Ho Bum;Ryoo, Seung Heui;Lee, Sang Hoon;Jeon, Byung Soon;Sang, Byung Chan
Korean Journal of Agricultural Science
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v.25
no.1
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pp.79-88
/
1998
This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.
In Korea, the urban studies of geography have mainly dealt with such a series of research as system of urban place and internal structure of urban area. The existing studies have been carried out with ecological approach. Ecologists, now a days, regard organiation and transfor-mation of the urban space as the process of invasion, succession, and segregation. However it is more proper that cities should be considered not as fragmantary objects, as some ecologists insist, but as synthetic ones in social structure. This research, with adopting a case of the renewasl of central area in Seoul, tried to make it clear that the formation and transition of the city is a product of social structure and examined polical and economic logic which exists in variation of urban space in detail. The results of this study are as follows; Urban renewal of central area is closely related with production and reproduction in capitalist society. In urban center, as business activities had increased since 1973 due to decen-tralization of production process, the necessity of reorganizing the land use in existing central area accordingly increased. The urban renewal program of central area in Seoul was inrroduced under such situation. The urban renewal of central area reflecting the capital logic has changed the central area with six hundred year's tradition. From the urban renewal of central area, not only was the central area, which traditionally had been mixed with various fun-ctions, simplified into the unitary area of busi-ness, but also physical landscape changed. As the land lot in renewal area expanded into regular shape, buildings became larger and taller. The program tremendously raised the price of related area. Aiming at these profits caused by the raised price, a great number of capitalists participated in the program. And as the benefit ratio of the manufacture sector continuously dropped with the economic recession, the pro-gram was carried out much more vigorously. That was because the idle capital accumulated during the recession was invested in property sector and was self-proliferated. The urban renewal raised the land value of central area and drove out the people living in this area. The people moved into the whole parts of the city resulting diffused squatter settlements. And the urban changes in central area were results of the policy of municipal authorities, who supported and systematized the changes lawfully and administratively, as well as reali-zation of capital logic. Due to the renewal policies of central area in Seoul, much more renewals by the only capitalists were carried out than those by the people themselves living in that area. The integration of land ownership in the law of urban renewal shows the reason of that. Moreover, the law allows the third deve-loper to participate in the tasks and admits the land expropriation rights. The municipal autho-rities guaranteed the profitability of the tasks through finacial aid, tax benifit, and relaxation of regulations for construction. As examined above, the changes in the land use of urban space have been led not by the ecological process of development of the city itself, but by the restructuring of capitalism and the intervention of the government authorities.
Kim, Cheol-Min;Park, Seung-Kyu;Shon, Mal-Hyun;Song, Sun-Dae;Kim, Young;Jun, Eun-Sook;Son, Han-Chul;Jung, Byung-Sun
Tuberculosis and Respiratory Diseases
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v.43
no.1
/
pp.30-37
/
1996
Background: The extraction methods of DNA from clinical samples are the major obstacle to use the PCR(polymerase Chain Reaction) in routine labortary for early detection of M. tuberculosis. We tried to improve the extraction method of DNA from sputum for establishment of the PCR in routine labortary by reducing the possibility of cross contamination and performing it easily and safely. Methods: We used the $InstaGene^{TM}$ DNA extraction kit(BioRad Co.) using Chelex 100 ion exchange resin for preparation of DNA. We compared InstaGene method in 100 cases of sputum from proteinase K method which is known as the most commonly used method for DNA purification(Experiment 1). And we compared InstaGene method in 98 cases of sputum from Microwave method developed by a company in Korea(Experiment 2). In experiment 1,245bps of IS6110 were amplified and then 188bps were amplified by nested PCR. In experiment 2,536bps in primary PCR and 276bps in nested PCR were amplified and analysed by agarose gel electrophoresis and EtBr staining. Results: When we chose AFB smear, culture, or AFB smear and culture as a standard test, PCR had low specificity and positive predictive value in both experiments. The InstaGene method has higher value in sensitivity and negative predictive value significantly than proteinase K method. The InstaGene method and the Microwave methods were similar in sensitivity, specificity, positive predictive value and negative predictive value. Conclusion: Even though both methods had lower possibility of cross contamination, shorter time requirement, simplicity, and economic advantages than Proteinase K method, the InstaGene method was a little simpler than the Microwave method. Therefore, in terms of usefulness in clinical application, the Instagene method seems to be the most useful method in DNA extraction for detection of M. tuberculosis using PCR. The reliability of this method will be clarified by further studies with enough clinical samples.
Journal of the Korean Society of Food Science and Nutrition
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v.41
no.12
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pp.1708-1715
/
2012
The purpose of this study was to investigate whether water extracts of Sparassis crispa (SC) have anti-obesity effects. Treatment of mature adipocytes with SC caused a decrease in lipid accumulation (assessed by Oil Red O staining) and an increase in glycerol release. Mice were induced to obesity by a high fat diet (45% fat in total kcal) and experimental groups were treated with two different dosages of SC extracts, a low SC (LSC, 100 mg/kg/day) or high SC (HSC, 300 mg/kg/day). SC extracts were administered by gavages for 10 weeks in the experimental groups, while the control group was fed with distilled water. The body weight gain of mice fed SC was significantly reduced (11.88% lower in LSC, 14.54% lower in HSC) compared to the control group. Additionally, there were significantly reduced serum levels of triglycerides (13.57% lower in LSC, 19.46% lower in HSC), total cholesterol (32.22% lower in LSC, 24.67% lower in HSC) and glucose (28.85% lower in LSC, 25.82% lower in HSC) in mice fed SC compared to the control group. Hepatic triglycerides in mice fed SC were lower (9.68% lower in LSC, 14.24% lower in HSC) than the control group and total cholesterol levels were also lower in mice fed SC (38.72% lower in LSC, 35.20% in HSC). These results demonstrate that the water extract of SC may enhance lipolysis and up-regulate the expression of lipolytic enzymes in vitro and reduce body weight in vivo. These significant effects were found for both low and high doses of SC treatment, and suggest SC can be used as potential therapeutic substances for the prevention and treatment of obesity.
It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.
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