• Title/Summary/Keyword: 정제 부분

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Cloning of the β-Lactamase Gene from Bacillus sp. J105 and Analysis of Its Expression in E. colis Cells (Bacillus sp. J105 유래 β-lactamase 유전자의 cloning 및 E. coli 내에서의 발현 분석)

  • Kang, Won-Dae;Lim, Hak-Seo;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Kwon-Il;Choi, Yung-Hyun;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1592-1599
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    • 2008
  • 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.

Issues and Considerations surrounding Revocation Physician's Medical License Arising from Criminal Offenses (의사의 형사범죄에 따른 면허취소처분의 쟁점과 고려사항)

  • Kim, Sung-eun
    • The Korean Society of Law and Medicine
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    • v.19 no.1
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    • pp.113-142
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    • 2018
  • In recent years, there have been opinions in which physicians are liable to the revocation of their medical license if they are sentenced to above a certain level for criminal charges regardless of the types of offenses. Accordingly, a revised bill of law was submitted in the National Assembly, and related discussions are thus expected to commence. Considering the morality and ethics or the level of the rule of law that the general public expects of physicians, as well as the license revocation system in other professional sectors, it is assessed that medical license revocation due to criminal convictions of physicians is appropriate to some degree. However, if a poorly devised system is established based on unrefined inferences or emotional judgements, unexpected side-effects are likely to arise. With regard to serious criminal acts that society generally perceives as unacceptable, it can be assessed that the revocation of physicians' licenses would appropriately protect the general public from threats. However, given the life-saving characteristics of high-risk medical practices, higher malpractice exposures, and social values, it is difficult to assess charges of professional negligence resulting in death(or in injury) and minor offences in the same manner as anti-social criminal offences are handled. Physicians need to be treated the same as any other professions. At the same time, they are engaged in administering medical treatment to patients in the face of great risks as professionals. Under the circumstances, a discussion on the introduction of a more specific and empirical system is needed by considering the intrinsic characteristics of medical treatment and the need for an equitable health and medical policy. Accordingly, based on the above judgment and perception, this study explores the code of ethics for physicians and medical license revocation related to criminal offences at home and abroad, and examines various legislative alternatives appropriate for the Republic of Korea. In doing so, the purpose of the study is to contribute to the development of a reasonable system for handling criminal offences by physicians.

Further Studies on the Specificity of the N- and C-terminal Antigenic Determinant of Hen Egg-white Lysozyme (계난백(鷄卵白) Lysozyme의 N-말단(末端)과 C-말단(末端) 항원결정기(抗原決定基)에 대한 연구(硏究))

  • Ha, Youn-Mun
    • The Journal of the Korean Society for Microbiology
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    • v.12 no.1
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    • pp.19-32
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    • 1977
  • The specificity of the N- and C-terminal antigenic determinant($P_{17}$: sequence $Lys^1-{cys-}^6-Asn^{27},\;{Trp^{12}}_2-Cys^{127}-Leu^{129}$) of hen egg-white lysozyme(HL) was studied in more detail. In a Scatchard plot of the binding of $^{14}C$-acetyl HL with guinea pig purified anti-$P_{17}$ antibody experimental values bent sharply aear r=1. This suggests of two antibody populations with different affinities for HL or possible steric hindrance in the binding of a second HL molecule to the second binding site of the antibody molecule. The antigenic activities of various peptides were tested by measuring their inhibition of the binding of $^{14}C-acetyl-P_{17}$ with the antibody, Only $P_{17}$ and $P_{17}t$(sequence $Lys^1-cys^6-Homoser^{12},\;Trp^{123}-Cys^{127}-Leu^{128})$) were inhibitory, with $K_1$ values of $2.0{\times}10^4$ and $8.1{\times}10^3$, respectively. These results indicate that the direct binding site of $P_{17}$ to anti-$P_{17}$ antibody may be located in the terminal portion of $P_{17}$ (sequence $Lys^1-Cys^6-Homoser^{12},\;Trp^{123}-Cys^{127}-Leu^{129})$) while the rest of $P_{17}$ may be important in maintaining the conformation of this determinant. The single disulphide bond involved in this determinant is essential for manifestation of immunological activity.

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Contents and Fatty Acid Compositions in Fats Extracted from Ice Creams and Ice Cream-Related Products (아이스크림 제품류에 함유되어 있는 지방함량 및 지방산조성)

  • Shin, Min-Kyung;Oh, Hyun-Hee;Hwang, Keum-Taek
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.6
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    • pp.721-728
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    • 2006
  • The objective of the study was to analyze the contents and fatty acid compositions in the extracted fats from selected commercial ice creams and ice cream-related products. Seventy four ice creams and ice cream-related products were collected from local stores: 22 regular 'ice creams', 10 premium 'ice creams', 22 'ice milks', 4 'sherbets', 11 'non-milk-fat ice creams' and 5 'non-milk product ice creams'. Contents and fatty acid compositions of the fats in the ice creams and ice cream-related products were analyzed. Fat contents in regular 'ice creams', premium 'ice creams' and 'ice milks' were $5{\sim}11%,\;13{\sim}17%\;and\;2{\sim}10%$, respectively. 'Sherbets', 'non-milk-fat ice creams' and 'non-milk product ice creams' contained $2{\sim}7%,\;4{\sim}11%\;and\;1{\sim}2%$ fats, respectively. Fats extracted from 14 regular 'ice creams', all of the premium 'ice creams' and 11 'ice milks' contained $63{\sim}75%$ saturated fatty acids and $2{\sim}5%$ trans fatty acids. Their fatty acid compositions were similar to those in milks and butter. However, fats from 8 regular 'ice creams' and 11 'ice milks' contained $11{\sim}28%\;and\;11{\sim}34%$ lauric acid, respectively. Since these levels of lauric acid were 3 times more than in milk or butter, other fats along with milk fat might be used for manufacturing these' ice creams' and 'ice milks'. Out of these 19 products, only 5 products were labelled as 'coconut oil' or 'refined oil' as well as milk fat being used. Fats extracted from 'sherbets', 'non-milk-fat ice creams' and 'non-milk product ice creams' contained $81{\sim}92%,\;76{\sim}99%\;and\;84{\sim}99%$ saturated fatty acids, respectively. Lauric acid was the most abundant fatty acid in the fats of these products, being $33{\sim}34%,\;17{\sim}45%\;and\;27{\sim}46%$ of the total fatty acids, respectively.

Studies on the Ginseng Plants(III) -Radioactive Sodium $Acetate-U-C^{14}$ Feeding Experiments- (인삼식물(人參植物)에 관한 연구(III) -동위원소화합물(洞位元素化合物) Sodium $Acetate-U-C^{14}$을 투여한 실험-)

  • Kim, Jung-Yun;Staba, E. John
    • Korean Journal of Pharmacognosy
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    • v.5 no.2
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    • pp.111-124
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    • 1974
  • The radioactive compound sodium $acetate-U-C^{14}\;(C^{14}-acetate)$ was administered to two- and four-year-old July and September American ginseng (Araliaceae, Panax quinquefolium L.) plants and cuttings. The $C^{14}-acetate$ uptake was approximately 99%. The autoradiochromatograms suggest that the saponins isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration (% plant dry weight) of semi-purified saponins were high in the leaves (13.8%), as compared to fruits (9.8%), stems (7.9%) and roots (6.3%). The average percentage of $C^{14}-acetate$ incorporation into panaquilins was 4.8%. The average percentage of $C^{14}-acetate$ incorporation into panaquilins B and C was higher (1.40% and 1.13%, respectively) than that into panaquilins C, (d), G-1 and G-2 (0.75%, 0.65%, 0.13% and 0.53%, respectively). Panaquilin synthesis may be depending upon the part, collection period and age of the plant. The average percentage of $C^{14}-acetate$ incorporation into panaquilin B is high in roots (0.58%) and stems (0.48%); that into panaquilins C and (d) high in leaves (0.40% and 0.45%, respectively); and that into panaquilin E high in roots and leaves (0.55% and 0.50%, respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-1). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-1 may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and cailus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that $C^{14}-acetate$ was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act. 0.56 mmcCi/mg) and four-year-old plants $(sp.\;act.\;0.54\;m{\mu}Ci/mg)$.

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Effects of Isoflavone Supplementation on Lipid Profiles and Antioxidant Systems in Rats Fed with Cholesterol Diet (고콜레스테롤 식이를 섭취한 흰쥐에서 이소플라본 보충이 혈청 지질패턴 및 항산화체계에 미치는 영향)

  • Kim, Soo-Yeon;Kim, Soon-Young;Chung, Chung-Eun;Yoon, Sun;Park, Jung-Hwa
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1683-1690
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    • 2010
  • There is an increasing interest in the potential of isoflavone in reducing the risk of cardiovascular disease and cancer, however, although several effects of isoflavone as a component of soy protein are well established, the hypolipidemic and antioxidant effects of purified isoflavone are still controversial. This study was to investigate the effects of isoflavone on serum lipid profiles and antioxidant status in rats. 7-week old male Sprague Dawley rats were fed one of the following diets for 8 weeks: basal diet (B), basal+0.3% isoflavone (BI), basal+0.5% cholesterol (BC), or basal+0.3% isoflavone +0.5% cholesterol (BIC). Two-way ANOVA was used to test the effects of dietary isoflavone and cholesterol supplementation and their interaction on variables. Serum lipid profiles and total antioxidant status (TAS) were examined spectrophotometrically. Degree of serum lipid peroxidation was measured by malondialdehyde (MDA) assay. The activities of serum antioxidant enzymes (GSH-Px, total-SOD) was determined. Levels of serum total cholesterol, VLDL+LDL-cholesterol and Atherogenic index were significantly lower in BI than those levels in group B (p=0.0002, p<0.0001, and p=0.0042, respectively). Serum total antioxidant status (TAS) levels were significantly higher, in both isoflavone supplemented groups (BI, BIC) compared to those levels in each control group (B, BC) (p<0.0001). Activity of total-SOD was significantly higher in BI compared to the activities in group B (p=0.0317). There was no interaction between isoflavone and cholesterol supplementation. In conclusion, isoflavone supplementation showed positive effects on the serum lipid profiles and total antioxidant activities in both conditions, either when fed a diet with or without cholesterol. These effects of isoflavone were independent of cholesterol supplementation.

Sensory quality, antioxidant, and inhibitory activities of XO and AO of Smilax china leaf tea fermented by Aspergillus oryzae (Aspergillus oryzae 발효 청미래덩굴잎 분말차의 관능적 품질 및 항산화능과 xanthine oxidase 및 aldehyde oxidase 저해활성)

  • Lee, Sang-Il;Lee, Ye-Kyung;Kim, Soon-Dong;Yang, Seung Hwan;Suh, Joo-Won
    • Food Science and Preservation
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    • v.21 no.1
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    • pp.129-139
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    • 2014
  • This study was conducted in order to investigate the optimal fermentation periods of the Smilax china L. leaves as a fermented tea via Aspergillus oryzae for 0 (non-fermented), and 10, 20, and 30 days (NF, F10, F20, F30). It was also observed for its quality characteristics. In the color and spectrum (400~700nm) of 1% tea water extract, NF was light yellow, whereas fermented tea (F10~F30) was light red color, and the F10 among F10~F30 has the clearest color and spectrum. Furthermore, acceptabilities of aroma and brightness were insignificantly different between NF and F10~30, while the mouth feel and overall acceptabilities were insignificantly distinct among all of the fermented teas. Therefore, these results suggest that the appropriate fermentation period for tea fermentation is 10 days. On the other hand, the total polyphenol and flavonoid content in the NF was the highest among all of the fermented teas. In the antioxidant parameters, EDA (electron donating ability), FRAP (ferric reducing antioxidant power), and LPOIA (lipid peroxidation inhibitory activity) in the NF were the highest among all fermented teas. Meanwhile, the XOI (xanthine oxidase inhibitory activity) was low, as well as insignificantly different from NF and F10~F30, whereas the AOI (aldehyde oxidase inhibitory activity) was markedly higher (38.09~41.70%) by the hot water tea extract (with or without fermentation), particularly the AOI that has increased via fermentation. In conclusion, the overall antioxidant activity tended to be reduced by fermentation; however, the EDA, FRAP and LPOIA in the fermented tea for 10 days was higher than the activities during 20~30 days of fermentation. There was a similar result in the color and acceptability of fermented tea for 10 days, which was remarkably better than those of 20-30 days. Therefore, fermented tea from the leaves of Smilax china L. could be expected to be used as a functional tea without the loss of inhibitory activity of both the XO and AO via fermentation.

Cryopreservation of Primordial Germ Cells(PGCs) from Korean Native Chicken(Ogye) Embryos using Commercial Cryoprotectants (상업용 동결보호제를 이용한 한국재래닭(오계) 원시생식세포의 동결 보존)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Do, Yoon Jung;Kim, Jae Hwan;Kim, Young Sin;Seong, Hwan Hoo;Ko, Yeoung Gyu;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.163-169
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    • 2013
  • Cryopreservation of poultry semen has been reported, but preservation of female genetic material has not been possible because of the unique anatomical and physiological characteristics of the avian egg. Thus an alternative strategy for conservation of oviparous species of animals must be developed. Recent technological developments for producing germline chimeras by the transfer of primordial germ cells (PGCs) into recipient embryos has enabled the conservation and retrieval of chicken genetic resources in their complete form. In the present study, fertilized eggs were incubated for about 5.5 days to obtain embryos at stage 28. The whole embryo was collected from the germinal gonad using a fine glass micro pipette under a microscope. The PGCs were then purified using MACS method. Two commercially available cryoprotectants (A and B) were used to preserve the PGCs, and EG were used as a control. The average recovery rate of PGCs after thawing was 35.5% and 60.5% with the A and B treatments, respectively. There was no significant difference between B treatments and control, which showed an average recovery rate of 52.8%. However, the recovery rate obtained using A cryoprotectant (35.5%) was significantly lower than using treatment control and B. The average viability of the PGCs after thawing were 77.9% and 77.4% for cryoprotectants A and B, respectively, and the control were was 81.6%. There was no statistically significant difference between the two treatments and control. It was concluded that all of the available cryoprotectants examined in this study could be used for preservation of PGCs from embryos. Further experiments to produce germline chimera from PGCs preserved using this techniques are strongly recommended.

Antioxidative, Antimicrobial, and Anti-proliferative Activities of the Floret and Stalk of Broccoli (Brassica oleracea L.) (브로콜리 꽃송이 및 줄기의 항산화, 항균 및 대장암 세포 생육억제효과)

  • Kim, Mi-Sun;Lee, Ye-Seul;Kwon, Ha-Young;Kim, Jong Sik;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.58-66
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    • 2014
  • In the course of study for a use for non-edible parts of broccoli (Brassica oleracea L), and the development of processed food utilizing these parts, edible floret and non-edible stalk were extracted with ethanol and different organic solvent fractions were prepared. With 10 different extracts and fractions, their useful components and various biological activities, such as antioxidant, antimicrobial and anti-proliferation activity, were investigated. The stalk has more abundant water soluble carbohydrate when compared with the floret, and floret has higher hexane-soluble pigments. Analysis of total flavonoid and total polyphenol contents showed that the floret has 1.5~1.99 times higher concentrations than the stalk. Among the fractions, ethylacetate (EA) fractions have the highest amount of total flavonoid and total polyphenol. The stalk and floret possessed 9.45 and 42.01 mg-total flavonoid/g, respectively. In the antioxidation activity assay, the EA fraction of floret showed strong radical scavenging activity and reducing power, while the n-hexane fraction of the stalk exhibited nitrite scavenging activity. In the antimicrobial activity assay, the EA fraction of floret showed a strong and broad-range of antibacterial activity, irrespective of gram positive or gram negative bacteria. In a while, the hexane and EA fractions revealed anti-proliferative effects against the human colorectal cancer cell HCT-116. Strong anti-proliferative activities were found in the hexane fraction of stalk (18.4% of cell viability), and the n-butanol fraction of floret (6.9% of cell viability). Our results suggest that the further study of the characterization of active fractions and the identification of active components from different parts of broccoli are needed to develop functional foods or novel plant-derived medicines.

Characterization of an Antarctic alkaline protease, a cold-active enzyme for laundry detergents (세탁세제 첨가용 효소 개발을 위한 남극 해양세균 유래 저온성 단백질분해효소의 특성 연구)

  • Park, Ha Ju;Han, Se Jong;Yim, Joung Han;Kim, Dockyu
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.60-68
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    • 2018
  • A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.