IBA 및 elicitor 처리에 따른 백수오 기내 생산 부정근 및 표준품의 FT-IR 스펙트럼 기반 대사체 비교 분석

안명숙,소은진,지은이,최소연,박상언,문병철,강영민,민성란,김석원,Ahn, Myung Suk,So, Eun Jin,Jie, Eun Yee,Choi, So Yeon,Park, Sang Un,Moon, Byeong Cheol,Kang, Young Min,Min, Sung Ran,Kim, Suk Weon 한국식물생명공학회 2018 식물생명공학회지 Vol.45 No.3

한약자원 약용부위와 대사체 수준에서 동등성이 규명된다면 식물조직배양을 통한 약용자원의 대량증식은 한약자원 약용부위의 보다 안전하고, 지속적인 대체 공급수단으로 활용이 가능하다. 따라서 본 연구에서는 기원 검증이 이루어진 백수오 약용부위와 동일조직에서 유래된 부정근으로부터 전세포추출물의 FT-IR스펙트럼 분석을 통해 대사체 수준에서의 동등성을 비교분석함으로써 한약자원 약용부위의 대체 공급수단으로 활용 가능성을 검증하고자 하였다. 식물생장조절제인 IBA 농도별 백수오 부정근의 생장률 분석 결과 5 mg/L IBA 첨가 배지에서 백수오 부정근 증식이 가장 우수하였다. SA 처리의 경우 처리 농도가 증가함에도 불구하고 백수오 부정근의 생장에는 차이가 없었지만, MeJA 처리시 농도가 높아질수록 부정근이 갈변되고 생장이 감소하였다. Elicitor 처리된 백수오 부정근 및 표준 약용부위 시료로부터 FT-IR 스펙트럼 데이터의 PCA 와 PLS-DA 분석결과 $400{\mu}M$ MeJA를 제외한 모든 처리구에서 백수오 기내 생산 부정근과 표준 약용부위 시료들 사이에서 전체 대사체 패턴이 매우 유사함을 알 수 있었다. 따라서 본 연구에서 확립된 FT-IR 스펙트럼기반 다변량통계분석 기술은 다양한 한약자원 약용부위 시료들의 대사체 수준 동등성을 신속하게 식별하는 수단으로 활용이 가능할 것으로 기대된다. To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared spectroscopy (FT-IR) can be used to discriminate and compare metabolic equivalence, standard medicinal parts of Cynanchum wilfordii (Maxim.) Hemsl. and their adventitious roots were subjected to FT-IR. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from FT-IR spectral data showed that whole metabolic pattern from the adventitious root of Cynanchum wilfordii was highly similar to its standard medicinal parts. These results clearly showed that mass proliferation of adventitious roots could be applied for the novel supply of standard medicinal parts of medicinal plants. Furthermore, FT-IR spectroscopy combined with multivariate analysis established in this study could be applied as an alternative tool for discriminating of whole metabolic equivalence from standard medicinal parts. Thus, it is proposed that these metabolic discrimination systems from the adventitious root of Cynanchum wilfordii could be applied for metabolic standardization of in vitro grown Cynanchum wilfordii.

Dimethylhydrazine으로 처리한 쥐에서 식이의 Calcium 함량과 지방종류에 따라 혈장 Cholesterol 수준과 Cholesterol 대사에 미치는 영향

박현서(Park Hyun-Suh),지은이(Jie Eun-Yee),강금지(Kang Keum-Jee) 韓國營養學會 1998 Journal of Nutrition and Health Vol.31 No.9

FT-IR 스펙트럼 데이터의 다변량 통계분석을 이용한 고기능성 아프리칸 얌 식별 및 기능성 성분 함량 예측 모델링

송승엽(Seung Yeob Song),지은이(Eun Yee Jie),안명숙(Myung Suk Ahn),김동진(Dong Jin Kim),김인중(In-Jung Kim),김석원(Suk Weon Kim) 한국원예학회 2014 원예과학기술지 Vol.32 No.1

본 연구에서는 UV-VIS spectrophotometer를 이용한 total carotenoids, flavonoids, phenolics 함량 데이터와 FT-IR 스펙트럼 데이터를 다변량통계분석법을 통하여 기능성 성분 함량이 높은 아프리칸 얌 고속 선발 시스템을 구축하였다. 62개 아프리칸 얌의 total carotenoids 함량은 0.01-0.91μg・g^-1 dry wt 나타냈다. Total flavonoids와 phenolics 함량은 12.9-229.0μg・g-1 dry wt와 0.29-5.2mg・g-1 dry wt로 각각 나타났다. 아프리칸 얌은 FT-IR 스펙트럼상의 1700-1500, 1500-1300, 1100-950cm^-1 부위에서 중요한 스펙트럼 변화가 나타났다. 이 부위는 각각 amide I과 II을 포함하는 아미노산 및 단백질계열의 화합물, phosphodiester group을 포함한 핵산 및 인지질 그리고 단당류나 복합 다당류를 포함하는 carbohydrates 계열의 화합물들의 질적, 양적 정보를 반영하는 부위이다. PCA 분석과 PLS-DA 분석에서 62개 아프리칸 얌은 유연성이 높은 종으로 3개의 그룹을 형성하였다. 아프리칸 얌의 FT-IR 스펙트럼 데이터와 UV-VIS spectrophotometer 을 이용한 total carotenoids, flavonoids, phenolics 함량 데이터 간에 PLS regression 분석하였다. Total carotenoids, flavonoids, phenolics 함량 성분의 실측 값과 예측 값간에 상관계수(R²)가 각각 0.83, 0.86, 0.72로 나타났다. 이 결과, 아프리칸 얌으로부터 FT-IR 스펙트럼을 이용한 total carotenoids, flavonoids, phenolics 함량 예측이 가능하였다. 본 연구에서 확립된 대사체 수준에서 아프리칸 얌의 유용 기능성 성분 함량 예측 모델링을 통해 품종, 계통의 신속한 선발 수단으로 활용이 가능할 것으로 예상된다. We established a high throughput screening system of African yam tuber lines which contain high contents of total carotenoids, flavonoids, and phenolic compounds using ultraviolet-visible (UV-VIS) spectroscopy and Fourier transform infrared (FT-IR) spectroscopy in combination with multivariate analysis. The total carotenoids contents from 62 African yam tubers varied from 0.01 to 0.91 μg・g^-1 dry weight (wt). The total flavonoids and phenolic compounds also varied from 12.9 to 229 μg・g^-1and from 0.29 to 5.2 mg・g^-1 dry wt. FT-IR spectra confirmed typical spectral differences between the frequency regions of 1,700-1,500, 1,500-1,300 and 1,100-950 cm^-1, respectively. These spectral regions were reflecting the quantitative and qualitative variations of amide I, II from amino acids and proteins (1,700-1,500 cm^-1), phosphodiester groups from nucleic acid and phospholipid (1,500-1,300 cm^-1) and carbohydrate compounds (1,100-950 cm^-1). Principal component analysis (PCA) and subsequent partial least squarediscriminant analysis (PLS-DA) were able to discriminate the 62 African yam tuber lines into three separate clusters corresponding to their taxonomic relationship. The quantitative prediction modeling of total carotenoids, flavonoids, and phenolic compounds from African yam tuber lines were established using partial least square regression algorithm from FT-IR spectra. The regression coefficients (R²) between predicted values and estimated values of total carotenoids, flavonoids and phenolic compounds were 0.83, 0.86, and 0.72, respectively. These results showed that quantitative predictions of total carotenoids, flavonoids, and phenolic compounds were possible from FT-IR spectra of African yam tuber lines with higher accuracy. Therefore we suggested that quantitative prediction system established in this study could be applied as a rapid selection tool for high yielding African yam lines.

B16F10 흑색종 세포에 대한 Biorenovation 기법을 이용한 파리풀 캘러스 추출물의 Melanin 생성 억제 효과

이경미(Kyung-Mi Lee),박태진(Taejin Park),지은이(Eun Yee Jie),김석원(Suk Weon Kim),한동환(Dong-Hwan Han),김승영(Seung-Young Kim) 한국생물공학회 2021 KSBB Journal Vol.36 No.1

Bio-renovation is the microorganism-assisted conversion of the structure and improvement of biological efficacies of natural products and synthetic compounds. In this study, we investigated the effect of plant callus on anti-melanogenic effect and compared the activity changes of bio-renovated callus. Phryma leptostachya bio-renovation products at the concentrations of 100, 200, 400 μg/mL suppressed tyrosinase activity and the expression of melanogensis-related protein such as tyrosinase-related protein-1 (TRP-1), tyrosinase-related protein-2 (TRP-2) and microphthalmia-associated transcription factor (MITF) without a loss of cell viability in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 melanoma cells relative to its substrate. Moreover, cell viabilities of PLBR increased over the tested concentrations compared to its substrate (PL). PLBR at 400 μg/mL inhibited the α-MSH-induced melanogensis effect to the level of non-treated control. These results indicate that the structure of various compounds contained in Phryma leptostachya (PL) were modified through a bio-renovation technique and which conferred anti-melanogenic properties to PL.

Establishment of Plant Regeneration System Via Somatic Embryogenesis and Shoot Organogenesis from Leaf, Petiole and Stem Explants of Gynura Procumbens

Suk Weon Kim(김석원),Nornita Sham(노니타),Eun Yee Jie(지은이),Myung Suk Ahn(안명숙),Seung A Choi(최승아),Ye In Jeon(전예인) 한국원예학회 2016 한국원예학회 학술발표요지 Vol.2016 No.10

쥐에서 dimethylhydrazine 처리와 식이지방 종류에 따라 HMG CoA reductase 활성도와 bile acid 및 중성 sterol 배설에 미치는 영향

김영현,박현서,지은이 한국지질학회 1999 韓國脂質學會誌 Vol.9 No.2

The study was designed to observe the effect of 1,2-dimethylhydrazine (DMH) and the type of dietary fat on plasma lipids, hepatic microsomal level of HMG-CoA reductase, membrane fluidity, fecal excretion of bile acids and neutral sterols in rats. Male Sprague Dawley rats, at 7 wks of age, were divided into 2 groups (control and DMH), and each group was subdivided into corn oil and perilla oil groups, and fed experimental diet for 20 wks. Each rat of DMH group was intramuscularly injected with DMH (total dose of 180 ㎎/㎏). Regardless of the type of dietary fat, DMH treatment had shown significant reduction in plasma cholesterol and TG levels. Perilla oil rich in n-3 α-linolenic acid significantly reduced plasma cholesterol level compared with corn oil rich in n-6 linoleic acid, but plasma TG level was not significantly reduced by perilla oil. There was no significant effect on hepatic microsomal level of cholesterol and HMG CoA reductase activity by DMH and dietary oil. Microsomal PL content was significantly increased by perilla oil, which resulted in the reduction of cholesterol/phospholipid (Chol/PL) ratio and increased membrane fluidity. DMH treatment significantly increased the excretion of secondary bile acid (doxycholic and lithocholic acid), but perilla oil significantly reduced its excretion. DMH treatment had shown the trends of increasing the excretion of neutral sterols, but perilla oil significantly decreased its excretion in DMH group. In conclusion, plasma cholesterol and TG levels were reduced by DMH treatment which might be from the increased requirement of nutrients for tumor growth. However, perilla oil could reduce fecal excretion of secondary bile acid (colon cancer promotor) which was increased by DMH, and also reduced plasma cholesterol level by increasing hepatic microsomal fluidity. Therefore, it could be recommended to use more perilla oil in meal preparation to prevent cardiovascular disease and colon cancer.