The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.
Kim, A Rang;Jung, Min Chul;Jeong, Hye In;Song, Dong Gi;Seo, Young Bin;Jeon, Young Hee;Park, So Hyun;Shin, Hyuk Soo;Lee, Sang Lae;Park, Soo Nam
Applied Chemistry for Engineering
/
v.29
no.2
/
pp.176-184
/
2018
In the present study, we investigated the antioxidative properties, cellular protective effects and component analyses of 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from Lysimachia christinae Hance (L. christinae Hance). In the evaluation of antioxidative properties, the free radical scavenging activities ($FSC_{50}$) of 50% ethanol extract, ethyl acetate fraction and aglycone fraction were 146.8, 22.2 and $27.2{\mu}g/mL$, respectively and total antioxidant capacities ($OSC_{50}$) were 29.3, 2.9 and $4.5{\mu}g/mL$, respectively. The ethyl acetate fraction showed the highest free radical scavenging activity and total antioxidant capacity. Also, the cellular protective effects (${\tau}_{50}$) of 50% ethanol extract, ethyl acetate fraction and aglycone fraction on $^1O_2$ induced photohemolysis of human erythrocytes were 26.9, 57.5 and 103.9 min at $5{\mu}g/mL$, respectively. In particular, ${\tau}_{50}$ of the aglycone fraction exhibited a higher cellular protective effect than that of (+)-${\alpha}$-tocopherol (37.7 min). The cell viability of the ethyl acetate fraction on the UVB-induced cell damage increased up to 90.1%. In addition, the ethyl acetate fraction ($5-25{\mu}g/mL$) showed cellular protective effects on the $H_2O_2-induced$ cell damages in a dose-dependent manner. TLC, HPLC, UV-vis spectroscopy and LC-MS were used to analyse components of the ethyl acetate fraction and the main components were quercetin, kaempferol and their glycosides. In conclusion, L. christinae Hance extract/fraction can function as antioxidants to protect the skin exposed to UV radiation and may also be used as a novel functional cosmetic material, for example, an antioxidant against skin photoaging.
This study evaluated the quality characteristics of rice-doenjang prepared with Aspergillus oryzae MD05 and Bacillus subtilis HJ18-4. The physicochemical characteristics of rice-doenjang prepared with three types of meju, such as grain type(Rice 1), disc shape(Rice 2), and brick shape(Rice 3), were compared. We determined amino and ammonia nitrogen contents, protease and amylase activities, reducing sugar content, and total aerobic counts and lactic acid bacteria counts. The amino nitrogen and reducing sugar contents in all meju types gradually increased during fermentation. The amino nitrogen contents reached 151.5~200.0 mg%. Rice 3 were had the highest levels among the 4 types of rice-doenjang. After 42 days of fermentation, amylase and protease activity of Rice 3 were 366.57 and 94.26 unit/g, respectively, which were the highest levels among the 4 types of rice-doenjang. The total aerobic bacteria counts were not changed during 42-day fermentation. This results suggest that high quality characteristics of brick-shaped meju(Rice 3) were better than those of the other meju preparations.
Periodontal ligament (PDL) cells have been known as multipotential cells, and as playing an important rolesin periodontal regeneration. The PDL cells are composed of heterogeneous cell populations which have the capacity to differentiate into either cementoblasts or osteoblasts, depending on needs and conditions. Therefore, PDL cells have the capacity to produce mineralized nodules in vitro in mineralization medium which include ascorbic acid, ${\beta}$-glycerophosphate and dexamethasone. In spite of these well-known osteoblast like properties of PDL cells, very little is known about the molecules involved in the formation of the mineralized nodules in the PDL cells. In the present study, we analysed gene-expression profiles during the mineralization process of cultured PDL cells by means of a cDNA microarray consisting of 3063 genes. Nodules of mineralized matrix were strongly stained with alizarin red S on the PDL cells cultured in the media with mineralization supplements. Among 3,063 genes analyzed, 35 were up-regulated more than two-fold at one or more time points in cells that developed matrix mineralization nodules, and 38 were down-regulated to less than half their normal level of expression. In accord with the morphological change we observed, several genes related to calcium-related or mineral metabolism were induced in PDL cells during osteogenesis, such as IGF-II and IGFBP-2. Proteogycan 1, fibulin-5, keratin 5, ,${\beta}$-actin, ${\alpha}$-smooth muscle actin and capping protein, and cytoskeleton and extracellular matrix proteins were up-regulated during mineralization. Several genes encoding proteins related to apoptosis weredifferentially expressed in PDL cells cultured in the medium containing mineralization supplements. Dkk-I and Nip3, which are apoptosis-inducing agents, were up-regulated, and Btf and TAXlBP1, which have an anti-apoptosis activity, were down-regulated during mineralization. Also periostin and S100 calciumbinding protein A4 were down-regulated during mineralization.
Seo, Jung-Soon;Lee, Jae-Sung;Byun, Gwang-In;Kwak, Eun-Jung
Journal of the Korean Society of Food Science and Nutrition
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v.37
no.11
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pp.1472-1478
/
2008
Four kinds of wild grape Yakju were fermented with wild grape and cereal such as rice, glutinous rice, barley or millet, and their quality characteristics were investigated. Reducing sugar, total sugar, and activities of $\alpha$- and $\beta$-amylase decreased until 2nd day, increased on 3rd day, and gradually decreased after 5 days of fermentation. Reducing sugar content was in the order of glutinous rice> barley> rice> millet group, and this was consistent with the result of $\beta$-amylase activity. Total sugar content was in the order of glutinous rice> rice> barley> millet group, and it was consistent with starch content of each cereal. Alcohol content decreased after 2 days, then glutinous rice group gradually increased from 3 days whereas other groups showed no difference from 5 days of fermentation. pH decreased until 2 days of fermentation, slightly increased and then showed almost no change after 5 days of fermentation. Total acid increased until 2 days, then decreased on 3rd day of fermentation, and gradually increased in the order of barley> millet> rice> glutinous rice group. L value was the highest in the initial day of fermentation, however it decreased with fermentation time. While a value was also the highest in the initial day of fermentation, it gradually decreased after slightly increasing in 3 days of fermentation. In contrast, b value was the lowest in the initial day of fermentation. It greatly increased until 3 days, then decreased on 5th day of fermentation and gradually increased again. Groups of rice and glutinous rice had more brown color than those of barley and millet. In the sensory evaluation, wild grape Yakju using rice was the most preferred in taste, flavor, and overall preference.
LEE Eung-Ho;PARK Hyang-Suk;OH Kwang-Soo;CHA Yong-Jun
Korean Journal of Fisheries and Aquatic Sciences
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v.18
no.4
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pp.316-324
/
1985
Vacuum-packed and seasoned smoked-dried products of red squid, Ommastrephes bartrami, caught in the Northern Pacific Ocean, were prepared and stored at room temperature for 90 days to test their keeping quality. Defrosted squids were eviscerated, skinned, and cut. The mantle meats were flavored with seasoning powders prepared from sugar, sorbitol, salt, monosodium glutamate, or smoke flavor (Smoke-EZ, Alpha Foods Co., Ltd.). After seasoning, the mantle meats were dried at $45^{\circ}C$ for 7 hours, vacuum packed in plastic film bags, and pasteurized in water at $95^{\circ}C$ for 30 minutes. Three kinds of products were prepared : control products (seasoned-dried), solid smoked seasoned-dried and liquid smoked seasoned-dried. The moisture level, water activity, color value (L, a and b value), texture, and viable cell counts of bacteria in these products were determined during storage at room temperature, $5^{\circ}C\;and\;35^{\circ}C$, respectively. The results showed that the products could be preserved at good condition for 90 days though they developed pale brown color during storage. The contents of free amino acids, nucleotides and their related compounds, and the compositions of fatty acids of raw squid and smoked products were analysed. In the amino acids, arginine, taurine, glycine and proline were abundant in raw and smoked products. The contents of hypoxanthine of raw and smoked products were higher than the other nucleotides and their related compounds. In fatty acid compositions of raw and smoked products, the dominant fatty acids were docosahexaenoic acid (22:6), hexadecanoic acid(16:0) and eicosapentaenoic acid (22:5).
This study was designed to investigate the relationship between the stages of exercise behavior change and physical self-concept and self-efficacy of security employees in hotel casinos. The sampling was drawn from employees at 8 casinos which had more than 30 employees. Participants were selected by convenience sampling method and they completed questionnaires about Physical Self-Concept and Self- Efficacy by self-administration method under supervision of trained researchers SPSS 16.0 (Statistical Package for the Social Science) was used for data analysis in the present study. Reliability and validity were examined for the present study. The principle component factor analysis and varimax rotation were used for the present study. Eigen value 1.0 was the criterion for selecting factors. Chi-square (X) 2 test was utilized for measuring the difference in gender and types of job duties at the stages of exercise behavior change. One-way ANOVA was employed to examine the relationship between the stages of exercise behavior change as an independent variable and physical self-concept and self-efficacy as dependent variables. The Scheffe method was used to determine mean differences of groups as a follow-up test. Multiple regression analysis was utilized to test the difference of physical self-concept as dependent variable and self-efficacy as independent variable. To verify hypothesis for the study, a statistical significance level of $\alpha$=.05 was used. The results were as follow: first, there were differences found for gender and types of job responsibilities in the stages of exercise behavior change. Secondly, as security employees progressed through the stages of exercise behavior change, their physical self-concept and self-efficacy improved. Finally, physical activity and body fat had significant main effects on self-efficacy.
Kim, Suk-Shin;Kim, Sang-Yong;Noh, Bong-Soo;Chang, Kyu-Seob
Korean Journal of Food Science and Technology
/
v.31
no.3
/
pp.619-624
/
1999
This work was to study the potential health food use of germinated colored rice after germinating and drying using microwave under vacuum. Colored rice was soaked in water at $15^{\circ}C$ for 2 days and then germinated at $25^{\circ}C$ for $3{\sim}4\;days$. The germinated colored rice was dried by different drying methods: microwave vacuum drying 1, microwave vacuum drying $2\;(drying{\rightarrow}crushing{\rightarrow}drying)$, hot air drying, vacuum drying and freeze drying. Each drier except freeze drier was set to maintain the sample temperature at $60^{\circ}C$. During microwave vacuum drying 1 or 2, the sample reached $60^{\circ}C$ much faster (within 5 min) and was dried much faster ($2{\sim}3\;hrs$ than the other drying methods. The initial drying rate of microwave vacuum drying was ten times faster than that of hot air drying. The microwave vacuum drying 2 retained the highest ${\alpha}-amylase$ activity, followed by microwave vacuum drying 1, freeze drying, vacuum drying, and hot air drying.
Ha, Eun-Suk;Hwang, Soo-Hyun;Shin, Kwang-Soon;Yu, Kwang-Won;Lee, Keyung-Ho;Choi, Joo-Sun;Park, Woo-Mun;Yoon, Taek-Joon
Korean Journal of Food Science and Technology
/
v.35
no.6
/
pp.1209-1215
/
2003
This study was carried out to establish a quantitative analysis method of separating immuno-activating substance (EN-SP) from Acanthopanax senticosus (A. senticosus) by competitive direct ELISA. Mouse antiserum (anti-EN-SP) against EN-SP was generated by immunization (s.c.) of EN-SP purified from A. senticosus as an immunogen. The titer of anti-EN-SP was about 1 : 400, and the optimal dilution of EN-SP-HRP conjugate was 1 : 1,000. When the standard curve was constructed by ELISA, its sensitivity was about $0.2{\mu}g/mL$. The coefficient variation of intra- and inter-assay were $6.13{\sim}8.81%$ and $6.73{\sim}8.60%$, respectively. According to the standard curve, the concentration of EN-SP in various senticosus extracts was found to be only $59.85\;{\mu}g$ in 10mg of extract from the bark of A. senticosus. Similarly, the immunostimulating activity to produce $TNF-{\alpha}$ or IL-12 among the various extracts of Acanthopanax was shown to be correlated with the content of EN-SP. These results demonstrated that competitive ELISA was a convenient, fast, reproducible, and accurate method for the determination of EN-SP as an immunologically active standard substance in extract of A. senticosus.
Hypericum ascyron has long been used as medicinal plant and recent studies reported that H. ascyron has anti-diabetic, anti-oxidant, and anti-bacterial effects. In this study, inhibitory effect from H. ascyron on pro-inflammatory responses has been investigated. H. ascyron was extracted at optimal extraction condition. Total phenolic contents in water and 90% ethanol were 29.75 and 31.82 mg/g, respectively. Hyaluronidase inhibitory activity of H. ascyron extracts ($50-200{\mu}g/mL$ phenolics) was 0.00-14.81% and 15.33-47.49%, respectively. In cell viability, cell toxicity was shown at concentration of $100{\mu}g/mL$ and $30{\mu}g/mL$ of water and 90% ethanol extract. Therefore, $10-50{\mu}g/mL$ in water extracts and $5-20{\mu}g/mL$ in ethanol extracts was selected each for further study. Inducible nitric oxide synthase (iNOS) derived nitric oxide (NO) and cyclooxygenase (COX)-2-derived prostaglandin $E_2$ ($PGE_2$) protein expression inhibitory effect of extracts were inhibited in a dose dependent manner, significantly. Also, the pro-inflammatory cytokines inhibitory effect such as tumor necrosis $factor-{\alpha}$, nterleukin (IL)-6 and $IL-1{\beta}$ were decreased in the dose dependent manner. The results indicate that H. ascyron extracts reduced inflammatory responses in lipopolysaccharide-induced 264.7 cells via the regulation of the iNOS, COX-2, NO, $PGE_2$, and pro-inflammatory cytokines. Therefore, H. ascyron extracts have significant anti-inflammatory effect and a source as therapeutic materials.
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