Bacillus subtilis JS-17이 생산하는 Collagenase의 정제 및 특성

임경숙,손승희,강호영,전홍기,Lim Kyoung-Suk,Son Shung-Hui,Kang Ho Young,Jun Hong-Ki 한국생명과학회 2005 생명과학회지 Vol.15 No.4

Collagnase는 천연 collagen의 triple-stranded helix를 분해할 수 있는 protease로서 조직의 수복과 재생 과정에서 collagen의 재형성과 세포의 이동에 아주 중요한 역할을 하고, 숙주 감염시에는 collagen 기질을 빠르게 분해함으로써 감염을 돕는다. 본 연구에서는 일반가정에서 식용하는 김치로부터 collagenase를 생산하는 균주를 분리하여 Bacillus subtilis로 동정하였으며 이를 Bacillus subtilis JS-17이라 명명하였다. Bacillus subtilis JS-17이 생산하는 collagenase의 최적 생산 조건은 $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4{\cdot}4H_2O,\;0.1\%\;citrate,\;0.1\%\;CaCl_2(pH\;7.0)$의 배지에서 $30^{\circ}C$, 200 rpm으로 72시간 동안 배양하는 것이다. 최적 조건에서 Bacillus subtilis JS-17이 생산하는 collagenase를 Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography, DEAE-Sephadex A-30 column chromatography를 거쳐 분리 정제하고, 얻어진 정제 효소의 특성에 대하여 검토하였다. 정제된 collagenase의 비활성은 growth medium에서 192.1 units/mg였고, $1.1\%$의 수율로 얻어졌으며 분자량은 28 kDa이었다. 정제된 collagenase는 $55^{\circ}C$까지는 $100\%$의 활성을 유지하였고 $65^{\circ}C$에서도 $60\%$ 정도의 활성을 유지하였다. 또한 pH $6.0\~9.8$에서 $60\%$ 이상의 활성을 유지하였다. 정제된 collagenase는 metalloprotease inhibitor인 EDTA와 O-phenanthroline에 의해 효소 활성이 감소하였을 뿐만 아니라 Ammoninum persulfate, L-cysteine, N-ethylmaleimide, SDS, $NaN_3$, NaF, $KMnO_4$, PMSF에 대해서도 활성이 감소하였다. 정제된 collagenase를 여러 가지 기질에 대해 효소 활성을 비교한 결과 collagen (type I)에 대해 기질 특이성을 가지고 있었다. Collagenases are generally defined as enzymes that are capable of degrading the polypeptide backbone of native collagen under conditions that do not denature the protein. An extracellular collagenase-producing bacterial strain was isolated from kimchi and identified to be Bacillus subtilis JS-17 through morphological, cultural, biochemical characteristics and 16S rDNA sequence analysis. Optimum culture condition of Bacillus subtilis JS-17 for the production of collagenase was $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4\cdot7H_2O,\;0.01\%\; MnSO_4\cdot4H_2O,\;,0.1\%$ citrate and $0.1\%\;CaCl_2$. The production of collagenase was optimal at $30^{\circ}C$ for 72 hr. A collagenase was isolated from the culture filtrate of Bacillus subtilis JS-17. The enzyme was purified using Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography and DEAE-Sephadex A-50 column chromatography The purified collagenase has an specific activity 192.1 units/mg. The molecular weight of the purified enzyme was estimated to be 28 kDa by SDS-PACE. The purified collagenase has $100\%$ activity up to $55^{\circ}C$.

녹두와 고구마전분의 온도에 따른 용출 아밀로오스 특성

임경숙,안승요 한국농화학회 1991 Applied Biological Chemistry (Appl Biol Chem) Vol.34 No.2

녹두와 고구마의 전분용액을 가열할 때 용출되는 아밀로오스의 특성에 대하여 살펴보았다. 아밀로오스 함량은 녹두가 25.9%, 고구마가 16.6 %였다. 용출되어 나오는 아밀로오스의 양은 온도가 상승함에 따라 모두 증가했으며 녹두에 있어서는 100℃ 에서 용출된 양이 전분의 22.7%였고 고구마는 20.6%였다. 이는 녹두로부터의 linear fraction의 용출이 더 많다는 것을 나타내며, 가용성 아밀로오스의 양이 많음을 보여준다. 한편 이를 총 아밀로오스의 양과 비교했을 때는 녹두의 경우 100℃에서 87.6%, 고구마는 123.8%로 나타났다. 분리한 아밀로오스에 대한 IBC와 β-amyloysis(%)를 실시한 결과 IBC의 경우 19∼20mg of 1/100mg of polysaccaride로서 순수하게 얻어졌음을 알 수 있었고 β-amylolysis (%)의 경우에는 시료간에 유의한 차는 없었으나, 온도에 따라 감소하는 양상을 보여 온도가 증가하면서 가지를 가진 아밀로오스가 혼합되어 나오는 것을 알 수 있었다. 분자량 분포를 보기 위한 겔 크로마토그래피를 실시한 결과, 두 전분 모두 온도가 증가하면서 분자량이 증가하는 경향을 나타내 온도가 증가하면서 보다 빠져나오기 어려운 가지를 갖는 큰 분자량의 아밀로오스의 용출이 일어나는 것을 알 수 있었으며, 녹두보다는 고구마가 큰 분자량의 아밀로오스의 비율이 높고 이는 온도가 증가할수록 현저하였다.

노인 영양개선 전략 연구 : 건강 관련 요인 및 영양위험지표 분석

임경숙,민영희,이태영 대한지역사회영양학회 1997 대한지역사회영양학회지 Vol.2 No.3

The elderly are the most rapidly growing segment of the population in Korea and the largest consumers of expensive medical care. It is reasonable to believe that improving the nutritional status would be beneficial to reducing morbidity and to impro- ving the quality of their lives. This study was conducted to assess the health status and the Nutritional Risk Index of the elderly, and to provide the basic data for the Elderly Nutrition Improvement Program at the Public Health Center. One hundred and forty seven(76 males. 71 females) aged 60 years and over living in Suwon, were assessed with anthropometric measurements, body fat analysis, blood pressure measurements, and a set of questionnaires about health and other related variables. Over half of them had less than a junior middle school education and were considered low income. The percentage of overweight and obese subjects was 33% by the BMI(Body Mass Index), and the prevalence of hypertension was 28$\%$(males) and 31$\%$(females). They had a lot of self-recognized health problems, the male elderly complained about more than the female elderly. In the case of psychological health status, however, the female elderly showed a higher proportions of depression than the male elderly(p$\ll$0.05). In social health status, the elderly had good relationships with friends and collegues, whereas they had poor relationships with their families. They had many nutritional risk factors, and smoking was the most prevalent risk factor for the male elderly and anemia was for the female elderly. The results of this study suggest that Elderly Nutrition Improvement Programs should be planned that can be easily followed. It would be helpful to design a program focusing on individual phychological and social health status, this would increase the efficiency of the program.

SHORT TERM EFFECTS OF BUTYLATED HYDROXYTOLUENE AND 2-ACETYLAMINOFLUORENE ON PEROXIDE METABOLISM IN RAT LIVERS

任景淑 水原大學校 1994 論文集 Vol.12 No.-

합성항산화제인 BHT와 간암유발물질인 2-AAF의 단기간 투여에 의한 쥐 간의 과산화물대사에 대해 살펴보았다. 단기간의 BHT 소량 투여(50mg/체중 kg)에 의해 쥐 간의 micosome 분획의 TBARS가 유의적으로 증가하였으나, 다량투여(100mg/체중 kg) 하거나 2-AAF와 함께 투여하면 TBARS는 유의적인 차이가 없었다. BHT와 2-AAF를 별도로 투여하면 과산화수소 생성이 변화가 없었으나, 같이 투여한 경우 유의하게 증가되었다. 이 때 cytosol 분획의 SOD 활성도는 2-AAF 에 의해 유의하게 감소하였으나, BHT 만을 투여하거나, BHT 와 같이 투여한 경우 유의적인 변화를 보이지 않았다. 따라서 단기간의 BHT 투여는 쥐 간의 과산화물 생성을 억제시키기보다는 오히려 증가시키는 것으로 보인다. The short-term effects of two levels of butylated hydroxytoluene(BHT) with 2-acetylaminofluorene(2-AAF) on hepatic peroxide metabolism were investigated. Short-term i.p. injection of lower BHT(50mg/body weight) exerted a significant increase of microsomal peroxidation estimated by thiobarbituric acid reactive substances(TBARS) levels, whereas higher level of BHT (100mg/kg body weight) of combined treatment of BHT and 2-AAF did not increase the lipid peroxidation. Microsomal hydrogen peroxide generation was significantly increased when BHT and 2-AAF treated together. SOD acivities of the cytosolic fraction was significantly decreased by single intraperitoneal injection of 2-AAF. LIVERS

서울시 생산자서비스의 공간적 분포특성에 관한 연구 : 금융 및 보험업 , 부동산업의 기업본사를 중심으로

임경숙 한국도시연구소 1997 도시연구 Vol.- No.3

TV 프로 시청에 있어서 아동이 지각한 사실에 관한 연구

임경숙,김혜숙 이화여자대학교 사범대학 시청각교육과 1972 시청각 교육 Vol.- No.6

학교급식 식단 분석

임경숙,이태영 대한영양사협회 1998 대한영양사협회 학술지 Vol.4 No.2

The menu planning is closely tied to the acceptability of the food and the satisfaction of the food service system to its client, therefore national school lunch program(NSLP) menu should consider the food preferences of the students. To provide appropriate information and guidelines about desirable NSLP menu planning, the comparison between the frequency of NSLP menus and food preferences of the students was performed. NSLP menus with cooking method of 776 meals were collected from 10 elementary NSLP schools in Seoul and Kyunggi province, and food preference data was surveyed with 1618 elementary school students from 4 NSLP schools in Korea in 1995. According to the cooking methods, NSLP menus served Kimchi(98.3%), rice(71.4%) and soup(71.1%) frequently. Based on the food groups, NSLP menus served vegetables most frequently(188%), followed by grains(101.4%), milk(100%), meat(57.3%), fruits(50.3%), and so on. By Spearman Rank Correlations analysis, the serving frequencies of vegetable side dishes(r=.33, p=.05) were positively correlated with the students' preference score. According to the cooking method, the serving frequencies of steamed dishes(r=1.0, p=.00), salads(r=1.0, p=.00), Kimchi(r=.85, p=.01) and rice one-dish meal (r=.80, p=.10) were positively correlated with the students' preference score. However, the serving frequencies of the main dishes(r=.13, p=.57) and soup group(r=.15, p=.57) were not significantly correlated with the students' preference score. Even though the serving frequency of fruit were negatively correlated with the students' preference score(r=-.80, p=.10). These results indicated that the NSLP menu planned the vegetable side dishes in accordance with the students' food preferences, but it planned the main dish, soup group, and meat side dish group discordantly. Therefore further efforts are needed to involve the preferred cooking methods for increasing the food acceptability. It could be used for the basic guidelines for menu planning of NSLP, and for the future improvement of NSLP in Korea.

일반학술발표 / 보건소 유아영양교육의 효과 평가

임경숙,한문화,김영주 한국영양학회 1999 추계학술대회초록 Vol.- No.-

中脘穴 艾灸治療의 文獻的 考察

林暻淑 圓光大學校大學院 1980 學位論叢 Vol.5 No.-

The results obtained by the study on the documentary records of the process of using moxa at Jung-Wan point in order to cure are as follows; 1. We can use moxibustion at Jung-Wan point with acupuncture. 2. The time required is from 5 minutes to 30 minutes and the number of times of moxibustion is from 3 times to 500 times. 3. Gyeong-Lag goes through five Gyeong…Im-Maeg, Su-Tae-Yang, Su-So-Yang, Jog-Yang-Kyeong, Su-So-Eum, and the beginning point of Su-Tae-Eum-pye-Gyeon and the terminal point of Jog-Gweol-Eum-Gan-Gyeong are connected to Jung-Wan again. 4. We can use acupuncture and moxibustion at Jung-Wan point in the cases of diseases of the digestive organs chiefly, diseases of the respiratory organ, mental diseases, diseases of the circulatory system, diseases of women and the nature of many other diseases.

초등학교 급식의 영양관리 현황 및 개선 방안 연구

임경숙,이태영,김초일,최경숙,이정희,권순주,김미옥 대한영양사협회 2004 대한영양사협회 학술지 Vol.10 No.2

The Korea School Lunch Program mandates that school meals should safeguard the health and well-being of our children. School meals provide a nutritious diet that contains at least 1/3 of the daily Recommended Dietary Allowances of necessary nutrients at reasonable prices. This study was performed to assess the nutritional management of primary school meals, and to provide basic data for improvement of school meal nutritional guidelines. Three hundred and thirty six dietitians who worked at primary school! were recruited using a stratified sampling method. A survey was done by e-mail with a set of questionnaires about nutritional meal management. During school lunch planning, dietitians referred nutrition standards with priority, followed by students' food preferences and food hygiene. They considered energy as a most important nutrient during primary school lunch planning, followed by protein and calcium. Protein is usually over-served, whereas iron, vitamin A and calcium are usually under-served during primary school meal planning. For first and second grade primary students, grains, meat and vegetable side dishes seemed to be large compared to their eating capacity. Over 92% of school dietitians agreed the needs of food group standards for primary school lunch program. For making a checklist of the school lunch program, energy, protein and calcium were highly recommended, and meat·fish·egg group, vegetable·fruit group and dairy group were also highly recommended by school dietitians. Cooking method was also highly recommended. School dietitians proposed that remodeling of school kitchens and new cooking machine were the most important factors for improving school lunch system. These results suggest that food standards should be established for health-oriented nutritional management of primary school lunch programs. Nutritional checklists could include nutrient-rich foods and food groups, and items about cooking methods.