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Expression, Purification and Functional and structural relationship of pyruvate dehydrogenase phosphatase

  • Kim, Young-Mi;Jung, Ki-Hwa
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2002.07a
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    • pp.236-236
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    • 2002
  • Pyruvate dehydrogenase phosphatase (PDP) is a mitochondrial protein serine/threonine phosphatase that catalyzes the dephosphorylation and concomitant reactivation of the pyruvate dehydrogenase componant of the pyruvate dehydrogenase complex (PDC). PDP consists of a Mg$\^$+2/ -dependent and Ca$\^$+2)-stimulated catalytic subunit (PDPc) of Mr 52,600 and a FAD-containing regulatory subunit (PDPr) of Mr 95.600. Catalytic subunit of pyruvate dehydrogenase phosphatase (PDPc) has been suggested to have three major functional domains such as dihydrolipoamide acetyltransferase(E$_2$)-binding domain, regulatory subunit of PDP(PDPr)-binding domain, and calcium-binding domain. In order to identify functional domains, recombinant catalytic subunit of pyruvate dehydrogenase phosphatase (rPDPc) was expressed in E. coli JM101 and purified to near homogeneity using the unique property of PDPc: PDPm binds to the inner lipoyl domain (L$_2$) of E$_2$ of pyruvate dehydrogenase complex (PDC) in the presence of Ca$\^$+2/, not under EGTA. PDPc was limited-proteolysed by trypsin, chymotrypsin, Arg-C, and elastase at pH7.0 and 30$^{\circ}C$ and N-terminal analysis of the fragment was done. Chymotrypsin, trypsin, and elastase made two major framents: N-terminal large fragment, approx. 50kD and C-terminal small fragment, approx. 0 kDa. Arg-C made three major fragments: N-terminal fragment, approx. 35 kD, and central fragment, approx. 15 kD, and C-terminal fragment, approx. 10 kD. This study strongly suggest that PDPc consists of three major functional domains. However, further study should be necessary to identify the functional role.

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The Optimization of Mevinolin Production by Medium Composition of Penicillium citrinum (Penicillium citrinum 의 배지 조성에 의한 Mevinolin 생산 최적화 연구)

  • 차월석;신성의;권규혁;김선일;이동병;이태범
    • KSBB Journal
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    • v.15 no.3
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    • pp.232-237
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    • 2000
  • Theses studies were made on the mevinolin production from Penicillium citrinum Thom (KCTC 6990) Culture conditions pH temperature carbon sources nitrogen sources mineral sources surfactants and glucose concentration were optimized. The results of glucose concentration and maximum mevinolin production according to incubating time in the flask nearly disappeared after 5 days and appeared after 7 days respectively. temperature and pH conditions of maximum mevinolin production were $24^{\circ}C$ and 3.7 pH respectively. The results of maximum mevinolin production according to the kind of nutrients were as follows. Glucose of carbon sources were 3.5 mg/L. Peptone of nitrogen sources were 3.5 mg/L TEX>$K_2HP0_4$ of mineral sources was 3.8 mg/L Tween 20 of surfactants were 4.5 mg/L Maximum mevinolin productioni of glucose con-centration was 4.0mg/L of glucose 100 g/L In the batch culture Maximum mevinolin concentration was 10.3 mg/L after 8 days. maximum mevinolin specific production rate 0.016 mg/g-hr. These results need to be studied more than ever about temperature pH 야ㅕㅡ and treatment of by-product oil in the batch culture and must do the fad batch from now to increase mevinolin productivity.

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The Effects on The Glass Processing by Alumina Addition in Soda Lime Glass (소다석회유리에서 Alumina첨가제에 따른 제병 공정의 영향)

  • Choi, Young-June;Kim, Jong-Ock;Kim, Taik-Nam
    • The Journal of Engineering Research
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    • v.4 no.1
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    • pp.69-85
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    • 2002
  • The chemical composition of bottle glass is consisted of Na2O-CaO-SiO2. However the cullet is mornally used in order to decrease the melting tsmperature. This induce the productivity of bottle and decreases the cost. The addition of plate glass decreases the Al2O3 content and in fluence the stone phenomenon and devitification in botle glass. Tus the Feldspar is added in order to increase the Al2O3 content when plate cullet was added in melting. The Tridymite crystal was observed over 7.5% Al2O3 contents, which shown as white crystal in appearance. It is Supposed that the Wollastonite Would be occurred in more over 7.5% Al2O3. This fad id well consised With the Litertctures.

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Purification and Characterization of Recombinant Acetohydroxyacid Synthase Catalytic Subunit in Haemophilus influenzae (Haemophilus influenzae의 Acetohydroxyacid Synthase Catalytic Subunit 재조합 단백질 발현 및 특성)

  • Noh, Kyoung-Mi;Choi, Kyoung-Jae;Park, Joon-Shik;Yoon, Moon-Young
    • Korean Journal of Microbiology
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    • v.43 no.1
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    • pp.19-22
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    • 2007
  • Acetohydroxyacid synthase (E.C.2.2.1.6., AHAS) is the enzyme that catalyses the first step in the synthesis of the branched-chain amino acids valine, leucine and isoleucine. The AHAS gene (TIGR access code HI2585) from Heamophilus influenzae was cloned into the bacterial expression vector pET-28a and expressed in the Escherichia coli strain BL21(DE3). The expressed enzyme was purified by $Ni^{2+}-charged$ HiTrap chelating HP column. The purified enzyme appears as a single band on SDS-PAGE with a molecular mass of about 63.9 kDa. The enzyme exhibits absolute dependence on the three cofactors FAD, $MgCl_{2}$ and thiamine diphosphate for activity. Specific activity of purified enzyme has 3.22 unit/mg and optimum activity in the pH 7.5 at $37^{\circ}C$. This enzyme activity has an effect on the buffer. When comparing the enzyme activity against the organic solvent, it followed in type and the difference it is but even from the aqueous solution where the organic solvent is included with the fact that the enzyme activity is maintained.

A Study on Expandibility of Contents Using 'Beauty and the Beast' (원작 '미녀와 야수'를 활용한 다양한 콘텐츠 확장성 연구)

  • Joo, Eun-Ryeong;Chung, Jean-Hun
    • Journal of Digital Convergence
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    • v.15 no.12
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    • pp.461-468
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    • 2017
  • 'Beauty and the Beast' was generated as a novel in 1740 and descended to date with the musical movie in 2017. This study is to understand what contents has been derived and expanded from one story for 277 years upon analyzing 'Beauty and the Beast' with the background of the times. Due to the limitation to include all the folktales and stories, the subjects of this study are limited with the contents that had been known and recorded as 'Beauty and the Beast.' 'Beauty and the Beast' has been expanded to 7 types of contents including novel, fairytale, movie, drama, animation, musical and film opera. It was created as a novel in the eighteenth century, made as a fairytale, and developed as a movie by Jean Cocteau, a movie director. With Industrial Revolution and the development of science, 'Beauty and the Beast' which derived into animation, drama, and musical contents, has been developed as the film opera thanks to the new trial of Philip Glass and the development of music industry. 'Beauty and the Beast' is still regenerating with a variety of contents continuously upon the development of computer technology and the fad of global remake. With anticipation of further analytical studies on 'Beauty and the Beast,' it is expected that the data of this study can be contributed as a reference in the other studies.

The Effect of Self-esteem, Academic Achievement and Family Functioning of Adolescents on Hope (일도시 청소년의 자아존중감, 학업성취수준, 가족기능이 희망에 미치는 영향)

  • Lee Jong Eun;Park Sun Nam;Park Ho Ran
    • Child Health Nursing Research
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    • v.7 no.1
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    • pp.74-84
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    • 2001
  • This study is the descriptive survey for the purpose of providing the basic data that establishes the strategy to promote adolescent's hope by the examining of self-esteem, academic achievement, family functioning and hope of adolescents and the investigating of the factors influencing the hope in adolescents. The subjects for this study were 456 students of the first and second year of man's senior high school that located in Seoul. The data were gathered from 16th to 31st of the October 2000. For the survey tool, it was used that the Family Assessment Device(FAD) of Epstein, Baldwin & Bishop(1983), the Self-Esteem Inventory(SEI) of Coopersmith(1975), the class record order and Hinds & Gatusso(1991)'s Hopefulness Scale for Adolescents(HSA). The collected data was analyzed by statistics methods as the descriptive and frequency analysis, t-test, ANOVA, Pearson and Spearman correlation coefficients and Stepwise multiple regression of SAS program. The results of this study were following : 1. The mean score of self-esteem of young people was 51.06±6.83 and the mean grade was 2.04. The high academic achievement was 29.2%, middle grade was 52.7%, and low grade was 18.1%. The mean score of the family functioning was 38.30±6.98 and the mean grade was 2.25. The mean score of hope was 84.26±16.45 and the mean grade was 3.51. 2. The hope in adolescents was significantly different according to their father's school career. The mean score of the group that the father's school career was below junior high school was 77.32. That was significantly lower than the mean score 84.59 of the group that the father's school career was above college and the mean score 85.18 of senior high school group(F=4.04, P= 0.0183). 3. The self-esteem was represented the positive correlation with family functioning(r=0.43) and the all of the 4 subscales(r=0.31, 0.41, 0.39, 0.30). And, it was highly ranked as much as family functioning was good. The academic achievement was represented the positive correlation with self-esteem(r= 0.15). Also, the positive correlation was shown between the affective responsiveness, role recognition and emotional support as the subscales of family functioning and academic achievement(r=0.11, 0.12). And so, academic achievement was high as much as self-esteem was high and affective responsiveness and role recognition and emotional support were good. The hope was represented positive correlation with self-esteem and academic achievement(r=0.42, 0.26), and with the whole of family functioning(r=0.15) and the 4 subscales(r=0.13, 0.16, 0.11, 0.13). So, hope was high as much as self-esteem was high, academic achievement was high and family functioning was good. 4. The influencing factors on the hope of adolescents were self-esteem(17.63%), academic achievement(3.41%), father's school career(0.84%). These factors made it possible to explain 21.88% of hope.

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Weight Reduction Dieting Survey and Satisfaction Degree and Diet Related Knowledge among Adult Women by Age (연령별 성인 여성의 체중감량 다이어트 실태와 만족도 및 관련지식)

  • Kim, Myung-Kyung;Lee, Gui-Chu
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.5
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    • pp.572-582
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    • 2006
  • This survey was conducted from June 2004 to August 2004, through questionnaires given to 432 women aged ranging from 20 to 50 years, who were living in the Seoul metropolitan area. The questionnaires were designed to determine the physique, level of obesity and dieting survey as well as to uncover which diets were in fashion and their satisfaction degree. Dieting-related knowledge and dieting side effects were also investigated. The results were as follows: In dieting survey, as the age increased, dieting for health reason (52.3%) increased, whereas that for aesthetic reasons (32.6%) decreased (p<0.05). The most effective methods of dieting were exercise (50.2%) and diet (45.0%) regimens. In regards to exercise frequency, 46.5% for none, 23.5% for 1-2 times a week, suggesting that most respondents do not practice exercise for dieting purposes (p<0.05). Major source for dieting was shown to be obtained from relative family or friends (31.7%), newspapers or magazines (29.3%) and internet (14.9%). As the age decreased, information from the latter increased, whereas that from TV program (21.2%) decreased (p<0.05). Reasons for excess weight were bad eating habits (39.6%), lack of exercise (38.3%), pregnancy and childbirth (10.6%). In fad dieting, while organic foods and aerobic exercises scored the highest satisfaction degree among diet and exercise regimens, on the other hand, sauna, steam baths and high temperature half-immersion baths scored the highest among surgery and other special therapies. As the age increased, the satisfaction degree for organic foods such as organic vegetable and high functional boiled cooked rice increased (p<0.05). As the age increased, scores on dieting-related knowledge decreased. Side effects of dieting included dizziness 06.2%) fatigue (15.1%) indigestion/constipation (11.1%) physical weakness 00.5%), loss of concentration (7.8%), dry skin (7.4%) loss of motivation (6.7%). The above results showed that attitude toward dieting among adult women varied with age and in light of the well-being trend, organic foods and aerobic exercises scored the highest satisfaction degree through all age.

A Study on the Effects of Selection Attributes for Agricultural Products on Using Local Food Store (농산물 구매선택 속성이 로컬푸드 직매장 이용에 미치는 영향 연구)

  • Chung, Joon-Ho;Hwang, Sung-Hyuk
    • Journal of Distribution Science
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    • v.14 no.4
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    • pp.117-125
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    • 2016
  • Purpose - As consumers' needs for purchasing fresh and safe food have been bigger in Korea, their interest in local food is also growing recently. So, the number of local food stores has been increased from 3 in 2012 to 103 in 2015. Local food stores should operate a business responding consumers' needs in order that local food stores are not to be a one-time fad. Therefore, the purpose of this study is to analyze the characteristics of consumers who use a local food store and provide helpful implications to design a strategy for sustainable growth of local food store. Research design, data, and methodology - In this study, Probit model was used for empirical analysis in order to examine the effect of purchase choice attributes of agricultural products, consumer's satisfaction, and their demographic factors upon the intention to use a local food store. After estimating coefficients of the probit model, marginal effects were calculated as a standard normal, and cumulative distribution is differentiated with respect to explanatory variables. To collect the data, questionnaire survey was carried out with the consumers using the local food store (Youngjin Nonghyup near to Jeonju city located in Jeollabuk-do). Result - The data analysis found that the more consumers are satisfied with local food store, the higher intention they have to use the local food store. In addition, it was known that the factors related to quality of agricultural products and shopping convenience among the purchase choice attributes have a considerable impact on the purchase intention of a local food store. In demographic factors, income was turned out to be an important factor affecting purchase intention of local food. Such a result supports the hypothesis that high income consumers are likely to purchase local food, which is based on the inference that consumers who have a high income tend to pursue wellbeing life. Futhermore, information delivery, through a reputable media source among general factors, was known to play an important role in forming an intention to purchase local food. According to the analysis of marginal effects, probability of purchase intention of a local food store is increased by 11.4%, if a monthly average income of a household is above 4.5 million Won(Korean currency). If purchasing satisfaction with local food stores is high, the probability of purchase intention would be increased by 24.1%. Likewise, such a probability goes up by 8.7%, 5.8%, respectively as an increasing one unit of quality of agricultural products and shopping convenience of local food stores, respectively. Conclusion - For attaining sustainable growth in a local food store, it is considered necessarily to establish a proper store operation system to meet consumers' needs, especially for quality and shopping convenience of local food. Moreover, as it was found that appropriate communication through media source has a positive effect on the intention to use local food store, PR activity seems to be necessary to expand the consumers' demands for local foods.

A Study of Analysis on Comparison of Laying Performance and Disease Occurrence of Welfare Approved Farms of Laying Hens with Housing Type (산란계 동물복지 인증농가의 사육형태와 품종별 생산성 및 질병발생 비교 분석 연구)

  • Hong, Eui-Chul;Kang, Hwan-Ku;Park, Ki-Tae;Jeon, Jin-Joo;Kim, Hyun-Soo;Park, Sung-Bok;Kim, Chan-Ho;Suh, Sang-Won;Kim, Sang-Ho
    • Korean Journal of Poultry Science
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    • v.44 no.2
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    • pp.143-149
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    • 2017
  • This study was carried out to collect basic data on the egg laying performance and disease occurrence on animal welfare-approved farms (AWAF) for laying hens with different housing types (windowless, open, and free-range house) and strains (Hy-line, Lohmann). Age at first egg laying was 151 d in the windowless house and 141 d in both the open and free-range houses, and the percentage peaks of each farm were 91.8%, 92.9%, and 86.3%. Average egg production according to housing types were 70.9%, 77.4%, and 65.6% at 18~38 wk old and 91.1%, 90.1%, and 76.2% at 32~36 wk. The average dirty and cracked egg ratio of the windowless, open, and free-range houses was 2.84%, 1.15%, and 0.23%, respectively. Mortality in the open house was lower than the other house types. Age of first egg laying for the Hy-line strain was 155 d and that for the Lohmann strain was 157 d, and the percentage peaks of the two strains were 92.9% (252 d) and 87.2% (237 d), respectively. Average egg production of the strains was 83.7% and 76.9%, respectively. The average dirty and cracked egg ratio of the Hy-Line and Lohmann strains was 3.85% and 2.97%, respectively. Mortality of the Hy-Line strain was lower than that of the Lohmann strain. There were significant differences (P<0.05) in the egg weight, HU, and egg yolk color of chickens raised in the open house compared with those of chickens raised in the other house types. In terms of antibody titer, that for Newcastle disease in open house chickens was lower than that of chickens raised in windowless and free-range houses. The antibody titer for low pathogenic avian influenza, avian infectious bronchitis, and avian pneumovirus was high at over 3.0 in all three farms. SG, Salmonella enteritidis, Salmonella pullorum, fowl adenovirus infection, and infectious laryngotrachitis were not detected in any of the three house types. The number of AWAF for laying hens has continuously increased in Korea, but there is currently little data on the housing types used on the AWAF for laying hens. Therefore, the results of the present study, in which analyzed the laying performance and disease occurrence on AWAF for laying hens, can use as basic data for the improvement of performance and prevention of disease on further AWFA for laying hens.

2020 Dietary Reference Intakes for Koreans: riboflavin (2020 한국인 영양소 섭취기준: 리보플라빈)

  • Lee, Jung Eun;Cho, Jin Ah;Kim, Ki Nam
    • Journal of Nutrition and Health
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    • v.55 no.3
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    • pp.321-329
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    • 2022
  • Riboflavin and its derivatives, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), are key components of mitochondrial energy metabolism and oxidation-reduction reactions. Proposed dietary reference intakes for Koreans (KDRIs), that is, estimated average requirements (EARs), for riboflavin, based on current knowledge of riboflavin and riboflavin derivative levels, and glutathione reductase activity, are 1.3 mg/d for men aged 19-64 years and 1.0 mg/d for women aged 19-64 years. By applying a coefficient of variance of 10%, reference nutrient intakes (RNIs) were set at 1.5 mg/d for men aged 19-64 years and 1.2 mg/d for women aged 19-64 years. Likewise, EARs and RNIs of riboflavin intake were proposed for all age groups and women in specific life stages such as pregnancy. Mean adult riboflavin intake for adults aged ≥ 19 years was 1.69 mg/d in Korea National Health and Nutrition Examination Survey (KNHANES) 2020, which was 124.9% of EAR according to the 2020 KDRIs. In the 2015-2017 KNHANES study, the mean riboflavin intake from foods and supplements was 2.79 mg/d for all age groups, and 32.7% of individuals consumed less riboflavin than EAR according to the 2020 KDRIs. For those that used supplements, mean intakes were 1.50 mg/d for riboflavin from foods, 10.26 mg/d from supplements, and 11.76 mg/d from food and supplements, and 5.5% of individuals consumed less riboflavin than EAR. Although the upper limit of riboflavin has not been established, the merits of increasing supplement use warrant further consideration. Also, additional epidemiologic and intervention studies are required to explore the role of riboflavin in the etiology of chronic diseases.