Streptococcus faecium M₇₄ · LC에 의한 L-α-Glycerophosphate Oxidase의 생산

이인애,이은숙,이준우,Lee, In-Ae,Lee, Eun-Sook,Lee, June-Woo 한국식품영양과학회 2003 한국식품영양과학회지 Vol.32 No.5

L- $\alpha$ -glycerophosphate oxidase(GPO)는 L- $\alpha$-glycero-phosphate를 산화시켜 dihydroxyacetone phosphate와 과산화수소의 생성을 촉매시켜 주는 효소로서 혈중에 존재하는 triglyceride의 양을 측정하는 kit를 개발하는데 이용할 수 있으며 본 연구 결과로 종균을 개발 및 확보하였으며, 이 종균의 사용으로 GPO의 발효공정기술을 확립하였고, 또한 이들 조효소로부터 순수한 효소를 생산할 수 있는 정제공정 기술을 확립하였다. 먼저 ATCC에서 네 가지 균주와 KCTC에서 세 가지 균주, 그리고 본 연구실에서 분리한 Streptococcus faecium $M_{74}$.LC 균주 등 8가지 균주의 성장상태를 비교하고 GPO의 역가를 측정 해 본 결과, 배양액 1 L당 ATCC 19634는 65 units, ATCC 12755는 60 units, S. faecium $M_{74}$. LC는 67 units로 S. faecium $M_{74}$.LC가 제일 높은 역가를 생산하였다. 또한 발효조에서 배지의 양을 3 L로 하여 배양온도는 $37^{\circ}C$, 교반속도는 300 rpm, 통기량은 0.5 L/min, 17시간 배양하였을 때 가장 많은 양의 균체가 생성되었으며, GPO의 생산량도 가장 많았다(256 units/L). 이 때 배지 조성은 glucose 0.1%, glycerol 0.2%, tryptone 1.0%, yeast extract 1.0% 및 $K_2HP0_4$0.5%로 하여 배양하였고 GPO의 정제공정은 염투석 분획과 이온 교환수지 공정으로 대별할 수 있으며, 이온교환수지는 DEAE-cellulose 칼럼을 이용한 정제수율은 약 47%를 얻을수 있었다. The objective of this study was to search for the best strain as a source of L- $\alpha$-glycerophosphate oxidase (GPO) production and to establish the process technology for the purification of GPO on an industrial scale. The GPO was produced by culturing Streptococcus faecium, and purified by ammonium sulfate, DEAE-cellulose and hydroxyapatite chromatography. The relative activity was 60 units/L for 5. faecim ATCC 12755, 65 units/L for 5. faecium ATCC 19634, and 67 units/L for 5. faecium $M_{74}$.LC, respectively. The optimum condition for fermentation was $37^{\circ}C$ for temperature, 300 rpm for stir rate, 0.5 L/min for aeration rate and 17 hours. The main culture medium prepared by the modified AC medium. AC medium consists of 0.1% glucose, 0.2% glycerol, 1.0% tryptone and 1.0% yeast extract, 0.5% $K_2HP0_4$, pH 7.0. The GPO was purified by ammonium sulfate fractionation and ion exchange column chromatography, The yield and purity were 17.2% and 5.3 fold, respectively.

Aspergillus 속 미생물에 의한 발효비지의 항비만 효과

이상일(Lee, Sang-Il),이예경(Lee, Ye-Kyung),김순동(Kim, Soon-Dong),임종환(Lim, Jong-Hwan),서주원(Seo, Ju-Won),이인애(Lee, In-Ae) 한국산학기술학회 2013 한국산학기술학회논문지 Vol.14 No.11

고지방식이로 비만을 유도한 ICR 생쥐를 동물 모델로 하여 7종의 Aspergillus속 미생물로 발효시킨 발효비지 의 항비만 효과를 조사하였다. 수컷 ICR 생쥐에 5주간 고지방 식이를 실시하여 비만을 유도하고 고지방식이에 발효 비지 2%를 첨가하여 6주 동안 사육하였으며, 체중, 섭취량, 혈청 총콜레스테롤, 중성지질, Alanine transaminase과 high-density lipoprotein cholesterol, 간성 glutathione과 lipid peroxide를 측정하였다. 체중의 경우에 AE4 발효 비지를 식이한 군에서 가장 많이 감소 (31.33%, P<0.05)하였고 간성 GSH와 ALT증가로 고지방혈증이 저해되었다. 이상의 결 과로부터 Aspergillus속 미생물로 발효시킨 발효비지는 항비만 효과가 있는 것으로 나타났으며, 나아가 인체에서도 유 의한 비만개선 작용을 기대할 수 있을 것으로 사료된다. The anti-obesity effect of soybean curd residues (biji) fermented by seven Aspergillus spp. was investigated with obese ICR mice fed a high-fat diet. After inducing obesity by feeding high-fat diet for 5 weeks, animals were fed with a high fat diet supplemented with 2% fermented soybean curd residues for 6 weeks. Total cholesterol, triglyceride, Alanine transaminase, high-density lipoprotein cholesterol, hepatic content of glutathione and lipid peroxide were determined. In the case of body weight, AE4 group showed most prominent decrease (31.33%, P<0.05) and increase of hepatic GSH and ALT demonstrated hyperlipidemia inhibition. From the results, it is concluded that soybean curd residues fermented by Aspergillus spp. has anti-obesity effect and it is thought that fermented soybean curd resides can reduce obesity in human significantly.

Monascus Pilosus로 발효시킨 검정콩 첨가 식이가 고지방식이 마우스의 체중과 혈청 지방함량 및 간 조직 항산화계 효소활성에 미치는 영향

이상일(Lee, Sang-Il),김순동(Kim, Soon-Dong),이예경(Lee, Ye-Kyung),김미정(Kim, Mee-Jung),이인애(Lee, In-Ae),최종근(Choi, Jongkeun),서주원(Suh, Joo-Won) 한국영양학회 2013 Journal of Nutrition and Health Vol.46 No.1

The anti-obesity effects of fermented black bean were tested with mice fed a high fat diet for seven weeks. Body weight gain and feed efficiency ratio (FER) in the high fat diet control (HC) group were markedly higher, compared with those of the normal control (NC) group, but were significantly lower in the 2% black bean powder supplemented high fat diet (BB) group and 2% black bean powder fermented by M. pilosus supplemented high fat diet (BBM) group, compared with those of the HC group. Food intake in the HC and BB groups was significantly lower than that of the NC and BBM groups. Water intake in the HC group was significantly lower than that of the NC group, but was higher in the BB and BBM groups, compared with that of the HC group. On the other hand, relative liver and kidney weight in the HC group was lower than that of the NC group, but was higher in the BB and BBM groups, compared with that of the HC group. In addition, whereas epididymal fat weight in the HC group was markedly higher than that of the NC group, it was significantly lower in the BB and BBM groups, compared with that of the HC group. Meanwhile, hepatic GSH in the HC group was significantly lower than that of the NC group, but was slightly higher in the BB and BBM groups, compared with that of the HC group. Although hepatic LPO in the HC group was dramatically higher than that of the NC group, it was significantly lower in the BB and BBM groups, compared with that of the HC group. In addition, serum TG, total cholesterol, and LDL-cholesterol in the HC group was significantly higher than that of the NC group, but was significantly lower in the BB and BBM groups, compared with that of the HC group. On the contrary, HDL-cholesterol in the HC group was significantly lower than that of the NC group, but was higher in the BB and BBM groups, compared with that of the HC group. In addition, activity of XOR D type in the HC group was lower than that of the NC group, but was slightly higher in the BB and BBM groups, compared with that of the NC group. Activities of ROS scavenging enzymes, such as SOD, GPX, and GST in the HC group were significantly lower than those of the NC group, but were significantly higher in the BB and BBM groups, compared with those of the HC group. In addition, serum ALT activity in the HC and BB groups was higher than that of the NC group, but was significantly lower in the BB and BBM groups, compared with that of the HC group. In histopathological findings, hepatic fat accumulation in the HC group was higher than that of the NC group, but was lower in the BBM group, compared with that of the HC and BB groups. In particular, antiobese, hypolipidemic, and antifatty liver effect of black bean powder fermented by M. pilosus was specifically higher than that of non-fermented steamed black bean. In conclusion, the constituents of black bean fermented by Monascus pilosus have been proven to not only inhibit obesity and hyperlipidemia but also decrease hepatic fat accumulation in high fat diet-induced obese mice. (Korean J Nutr 2013; 46(1): 5 ~ 14)

대장균군 검사용 간이 시험지 개발

이인애(In-Ae Lee),김재화(Jae-Wha Kim),이회구(Hee-Gu Lee),성창근(Chang-Keun Seong),최인성(In-Seong Choe),정태화(Tai-Wha Chung) 대한의생명과학회 1996 Biomedical Science Letters Vol.2 No.1

대장균군 검사용 간이 시험지는 본 실험실에서 국내 최초로 고안, 개발하였으며 이 간이 시험지법은 현장 검사법의 하나로 대장균군이 내는 succinic acid dehydrogenase 때문에 tetrazolium salt가 환원되어 적색 반점을 형성하는 것을 이용한 방법으로서 이 간이 시험지의 제조는 대체로 종래의 표준 평판법과 거의 동일한 조성의 배지와 사약을 사용하여 여지에 흡착치킨 후, 건조시켜(60℃) 멸균한 것으로 표준 평판법과 어떤 상관관계가 있는가를 검토하였다. 이 간이 시험지의 제조에서는 bile salt No. 3를 deoxycholate로 대체하여 제조 원가를 절감하였고, 또한 일본에서 현재 시판되고 있는 제품과 품질 비교시험을 하여 더 좋은 결과를 얻었으며 종래의 표준 평판법과 비교하였을 때도 오히려 표준 평판법(24-48시간 배양)보다 빠른 시간(16-20시간 배양)내에 판정할 수 있는 이점이 있으며, 표준 평판법에서는 없어서는 안될 배지나 배양 접시, pipette등의 자료 및 기구가 일체 필요없고 언제 어디서나 현장에서 직접 시험할 수 있어 매우 간편하며 또한 저렴한 가격으로 제조할 수 있는 경제성이 높은 이점을 갖고 있다. The objective of this study was to develop a paper strip which could determine E. coli qualitatively and quantitatively in water, wastewater, drinks, or food. This paper strip method was a simple and rapid test method that determine E. coli by visual identification. In this study, nutrient culture media were formulated and characterized for optimum conditions. Paper strips were then prepared by impregnating into the media and dried at 60℃. The test procedure is quite simple to use. The paper strip was dipped into a sample, and excess sample was removed. The strip was then incubated at 37℃ for 16 to 20 hours and the number of colonies on the strip was counted. The color of the colony spots produced by microorganisms varied depending on the media formulation. Violet-red spots were produced by E. coli. The test method was simple, rapid and no special laboratory equipment was necessary for visual identification. Therefore, this test method is applicable to on-site tests such as field tests or home tests. The paper strip method was compared with the standard agar plate method and Japanese commercial product. The method of the economical preparation of test strips was studied for production on industrial scale.

자가 음성평가와 체질량지수의 특성 비교

이인애(Lee, In-Ae),황영진(Hwang, Young-Jin) 한국산학기술학회 2013 한국산학기술학회논문지 Vol.14 No.3

본 연구에서는 자가 음성평가에 따른 체질량지수를 비교하고자 하였다. 제5기(2010년) 1차년도 국민건강영양 조사 이비인후과 검진을 완료한 19세 이상의 성인을 5,811명을 대상으로 하며 자가음성평가와 체질량지수의 연관성을 분석하였다. 또한 체질량지수와 연관성이 깊은 키, 몸무게, 허리둘레 등도 함께 비교하여 자가음성평가와 어떠한 상관 을 보이는지 살펴보았다. 비교를 위해 chi-square, t-test, 이변량 로지스틱 회귀분석을 이용하였으며 그 결과, 자가 음 성평가에 따른 체질량지수는 유의한 차이를 나타내는 특성을 보이며, 체질량지수가 과체중-2단계 비만인 경우 정상체 중보다 음성장애 위험률이 1.77배 높은 결과를 나타내었다. 또한 체질량지수가 음성장애 통제변수로서의 유의함을 확 인하였으며, 음성장애 진단 시 키와 몸무게를 통한 체질량지수를 함께 비교하는 것은 필요하다는 것이 확인되었다. The purpose of this study was to analyze the association between self-reported voice problem and body mass index. Data were collected from the 5th Korea National Health and Nutritional Examination Survey (2010) from 5,811 subjects(2,503 men and 3,308 women) aged 19 years and olders. chi-square, t-test and multi-nominal logistic regression analysis were used that to compare self-reported voice problem and variable(age, sex, hight, weight, waist measurement, body mass index). body mass index(OR=1.028, 95% CI: 1.003-1.056) was independently associated with self-reported voice problem(p<0.031). also over weight-two step obesity (OR=1.765, 95% CI: 1.036-3.006) were independently associated with self-reported voice problem(p<0.036). The results of comparison verified that body mass index are valuable self-reported voice problem of risk factor. when the evaluation were conducted, what was considered body mass index is needed.

Development of Modified Stokes Expression to Model the Behavior of Expanded Beds Containing Polydisperse Resins for Protein Adsorption

In-Ae Lee(이인애),Min-Ae Im(임민애),Keungarp Ryu(류근갑) 한국생물공학회 2004 한국생물공학회 학술대회 Vol.2004 No.10

모음에 따른 발성역치압력과 비성도 특성 비교 연구

이인애(Lee, In-Ae),황영진(Hwang, Young-Jin) 한국산학기술학회 2013 한국산학기술학회논문지 Vol.14 No.2

본 연구는 모음에 따라 발성역치압력과 비성도의 차이를 비교하고자 하였다. 2012년 5월부터 8월 동안에 대 학에 재학 중인 20~30대의 정상성인 여성 28명을 대상으로 실시하였다. 발성역치압력은 객관적인 공기역학적 검사도 구인 PAS(Phonatory aerodynamic system, model 6600, KAY electronics, Inc)를 이용하였으며, /p/와 세가지 모음 /a, i, u/를 각 5번씩 측정하여 가운데 3개를 비교하여 분석하였다. 또한 비성도는 NasometerΠ(model 6450, KAY electronics, Inc)를 이용하였으며, 세가지 모음 /a, i, u/를 5초 이상 연장발성하여 측정하였다. 그 결과, 모음에 따른 발 성역치압력은 유의한 차이를 나타내었으며, 모음에 따른 비성도도 유의한 차이를 보였다. 발성역치압력과 비성도간의 상관관계도 매우 높은 상관성을 나타내었다. 따라서 모음간의 발성역치압력은 연인두 폐쇄정도를 예측할 수 있으며 그에 따른 비성도와 매우 연관성이 깊음을 알 수 있었다. This study sought to compare the difference of Phonation Threshold Pressure and Nasalance according to vowels. We tested 28 normal females(20~30age) who be in attendance at a school from May to August 2012. We measured Phonation Threshold Pressure through PAS(Phonatory aerodynamic system, model 6600, KAY electronics, Inc) that aero-mechanical instrument called. Phonation Threshold Pressure was measured by using the consonant /p/ and the three vowels /a, i, u/ and was analyzed the three times of Phonation the five times. Also We measured Nasalance through the NasometerΠ(model 6450, KAY electronics, Inc). The vowel's phonation was recored by using the three vowels /a, i, u/. the nasalance score was measured via Nasometer. As a Result, Phonation Threshold Pressure according to three vowels show significantly difference. Also, Nasalance according to three vowels show significantly difference. Finally, Correlation coefficient of Phonation Threshold Pressure and Nasalance according to three vowels show positive relation ship. Therefore Phonation Threshold Pressure and Nasalance according to three vowels have closely relationship.

Cholesterol Oxidase 를 생성하는 토양 미생물의 분리 및 효소 생산에 관한 연구

이인애(In - Ae Lee),최용경(Yong - Kyung Choe),이홍수(Hong - Soo Lee),최인성(In - Seong Choe),정태화(Tai - Wha Chung) 한국미생물생명공학회 1992 한국미생물·생명공학회지 Vol.20 No.4

파킨슨병 환자와 정상노인의 모음 산출 특성 비교

이인애(Lee In Ae),김문정(Kim Moon Jeoung),황영진(Hwang Young Jin) 한국음성학회 2012 말소리와 음성과학 Vol.4 No.1

The purpose of this study is to compare the voice differences of persons with Idiopathic Parkinson's disease and a normal group according to five vowels. Eight persons with Idiopathic Parkinson's disease and a healthy control group of 22 were selected and every voice analyzed by MDVP. The first result showed that jitter measurements between the two group showed a significant statistical difference according to all vowels. Second, the two groups' shimmer measurements showed a significant statistical difference according to nearly all vowels. Third, jitter measurements between the five vowels were more relatively closely correlated persons with Idiopathic Parkinson's disease than the normal group. Fourth, shimmer figures between the five vowels more relatively closely correlated persons with Idiopathic Parkinson's disease than the normal group.

Streptococcus faecium M74ㆍLC에 의한 L-α - Glycerophosphate Oxidase의 생산

이인애(In-Ae Lee),이은숙(Eun-Sook Lee),이준우(June-Woo Lee) 한국식품영양과학회 2003 한국식품영양과학회지 Vol.32 No.5

L-α-glycerophosphate oxidase(GPO)는 L-α-glycerophosphate를 산화시켜 dihydroxyacetone phosphate와 과산화수소의 생성을 촉매시켜 주는 효소로서 혈중에 존재하는 triglyceride의 양을 측정하는 kit를 개발하는데 이용할 수 있으며 본 연구 결과로 종균을 개발 및 확보하였으며, 이 종균의 사용으로 GPO의 발효공정기술을 확립하였고, 또한 이들 조효소로부터 순수한 효소를 생산할 수 있는 정제공정 기술을 확립하였다. 먼저 ATCC에서 네 가지 균주와 KCTC에서세 가지 균주, 그리고 본 연구실에서 분리한 Streptococcus faecium M_(74)ㆍLC 균주 등 8가지 균주의 성장상태를 비교하고 GPO의 역가를 측정해 본 결과, 배양액 1 L당 ATCC 19634는 65 units, ATCC 12755는 60 units, S. faecium M_(74)ㆍLC는 67 units로 S. faecium M_(74)ㆍLC가 제일 높은 역가를 생산하였다. 또한 발효조에서 배지의 양을 3 L로 하여 배양온도는 37℃, 교반속도는 300 rpm, 통기량은 0.5 L/min, 17시간 배양하였을 때 가장 많은 양의 균체가 생성되었으며, GPO의 생산량도 가장 많았다(256 units/L). 이때 배지 조성은 glucose 0.1%, glycerol 0.2%, tryptone 1.0%, yeast extract 1.0% 및 K₂HPO₄ 0.5%로 하여 배양하였고 GPO의 정제공정은 염투석 분획과 이온 교환수지 공정으로 대별할 수 있으며, 이온교환수지는 DEAE-cellulose 칼럼을 이용한 정제수율은 약 47%를 얻을 수 있었다. The objective of this study was to search for the best strain as a source of L-α-glycerophosphate oxidase (GPO) production and to establish the process technology for the purification of GPO on an industrial scale. The GPO was produced by culturing Streptococcus faecium, and purified by ammonium sulfate, DEAE-cellulose and hydroxyapatite chromatography. The relative activity was 60 units/L for S. faecium ATCC 12755, 65 units/L for S. faecium ATCC 19634, and 67 units/L for S. faecium M_(74)ㆍLC, respectively. The optimum condition for fermentation was 37℃ for temperature, 300 rpm for stir rate, 0.5 L/min for aeration rate and 17 hours. The main culture medium prepared by the modified AC medium. AC medium consists of 0.1% glucose, 0.2% glycerol, 1.0% tryptone and 1.0% yeast extract, 0.5% K₂HPO₄, pH 7.0. The GPO was purified by ammonium sulfate fractionation and ion exchange column chromatography. The yield and purity were 17.2% and 5.3 fold, respectively.