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구름버섯으로부터 개미산 생성 효소의 분리 및 기능 분석
이수연 ( Su-yeon Lee ),장석윤 ( Seok Yoon Jang ),이수민 ( Soo-min Lee ),박미진 ( Mi-jin Park ),양지윤 ( Jiyoon Yang ) 한국목재공학회 2021 한국목재공학회 학술발표논문집 Vol.2021 No.2
지구온난화의 원인 물질로 알려져 있는 이산화탄소(Carbon dioxide, CO<sub>2</sub>)로부터 유용한 화합물로 전환하고자 하는 연구들이 시도되고 있으며 효소를 이용한 생물학적 전환 공정들이 보고되고 있다. Formate dehydrogenase(FDH)는 원래 개미산(formic acid)를 CO<sub>2</sub>로 산화시키는 효소로 알려져 있는데, 이 효소 반응의 가역성(환원반응)을 이용하여 CO<sub>2</sub>로부터 개미산을 생산하려는 연구가 진행되고 있다. 본 연구에서는 목재부후균 유래 FDH를 분리하여 이산화탄소로부터 개미산을 생성하고자 하였다. 목재부후균 중 백색부후균에 속하는 구름버섯(Trametes versicolor )을 국립산림과학원에서 분리하였으며 Internal Transcribed Spacer(ITS) 분석을 통해 종을 확인하였다. T. versicolor 균사로부터 FDH 유전자를 분리하기 위해 프라이머(Forword: 5’-TGTACTATGCTCACTTCCACCA-3’ and Reverse: 5’-CTCATCCCACGGTTACAGG-3’)를 사용하였고 pCR2.1-TOPO (Invitrogen, Carlsbad, CA, USA) 벡터에 sub-cloning하여 염기서열 확보한 결과 약 42kDa의 단백질 사이즈를 유추할 수 있었다. 이후 TvFDH의 발현을 위해 in vitro 상에서 반응이 가능한 무세포 단백질 합성 시스템(cell-free protein synthesis) 시스템을 이용하였다. pCR2.1-TOPO 복제 플라스미드 재조합된 TvFDH 유전자는 제한효소 BamH1, Not1으로 처리하였고 동일한 효소로 절단된 단밸질 발현벡터 pBIVT-N-terminal hexa histidine에 T4 DNA 리가아제를 이용하여 접합시켰다. 이렇게 재조합된 플라스미드는 cell-free protein synthesis kit(Bioneer, Korea)를 이용하여 단백질 발현을 시도하였다. Kit 매뉴얼에 따라 3시간 반응 후, 12% SDS-PAGE gel을 이용하여 단백질 발현을 확인하였다. 결과적으로 대조구와 비교하였을 때 TvFDH를 처리한 실험구에서 예상 단백질 사이즈(42kDa)의 발현량이 증가하는 것을 확인하였다. 현재 Ni-NAT 정제 과정을 거쳐 분리한 단백질을 이용하여 효소활성 및 개미산으로의 전환율을 확인 중에 있다.
Soo-Yeon Lee(이수연),Hye-Ji Jun(전혜지),Ji-Young Yoon(윤지영),Tae-Su Kim(김태수),So-i Park(박소이),Sung-Pyo Lee(이성표),Joo-Hoon Park(박주훈),Jin-Young Lee(이진영) 한국생명과학회 2012 생명과학회지 Vol.22 No.3
본 연구에서는 십자화과에 속하는 식물인 브로콜리를 적색광, 청색광, 적?청색광, 형광등을 이용해 건조시킨 광원별 브로콜리 에탄올추출물의 항산화, 미백 및 주름 개선 효과 등의 화장품약리활성을 검증하였다. 항산화 활성을 검증하기 위해 전자공여능, SOD 유사활성능, xanthine oxidase 저해활성능을 측정한 결과, 전자 공여능에서는 형광등, 청색광, 적색광, 적?청색광 브로콜리의 에탄올추출물의 순서로 높았고, SOD 유사활성능에서는 청색광이, xanthine oxidase 저해활성능에서는 청색광과 형광등의 브로콜리 에탄올추출물의 활성이 가장 높았다. 미백 효과를 검증하기 위해 tyrosinase 저해활성능을 측정한 결과, 적?청색광 브로콜리의 에탄올추출물이 가장 높은 활성을 나타내었고, 주름 개선 효과를 검증하기 위해 collagenase 저해활성능을 측정한 결과 청색광, 형광등, 적색광, 적?청색광 브로콜리 에탄올추출물 순으로 높은 활성을 나타내었다. 수렴 효과를 검증하기 위해 astringent 활성을 측정한 결과 형광등 브로콜리의 에탄올추출물이 가장 높은 활성을 나타내었음을 확인하였다. 따라서 적색광, 청색광, 적?청색광, 형광등을 이용해 건조시킨 광원별 브로콜리 에탄올 추출물의 기능성 소재의 가능성을 확인하였다. The purpose of this study was to research the cosmeceutical activity of 70% ethanol extracts when Brassica oleracea var. italica Plenck (BO) was dried under red lighting (RLD), blue lighting (BLD), red+blue lighting (RBLD), or white lighting (WLD). The electron-donating abilities of 70% ethanol extracts from drying BO under RLD, BLD, RBLD, and WLD were 62.8%, 68.1%, 60.9%, and 69.9%, respectively, at a concentration of 500 ppm. The superoxide dismutase (SOD)-like activities of 70% ethanol extracts from drying BO under RLD, BLD, RBLD, and WLD were 39.2%, 47.2%, 19.6%, and 21.6%, respectively, at a concentration of 1,000 ppm. In terms of xanthine oxidase inhibition effects, the proportions of 70% ethanol extracts after drying BO under RLD, BLD, RBLD, and WLD were 45.1%, 56.2%, 38.8%, and 53.3%, respectively, at a concentration of 1,000 ppm. The tyrosinase inhibition effects of 70% ethanol extracts when BO was dried under RLD, BLD, RBLD, and WLD were 21.0%, 13.1%, 26.0%, and 19.1%, respectively, at a concentration of 1,000 ppm. The collagenase inhibition effect of 70% ethanol extracts when BO was dried under RLD, BLD, RBLD, and WLD were 47.4%, 20.8%, 54.6%, and 37.2%, respectively, at a concentration of 1,000 ppm. The astringent inhibition effects of 70% ethanol extracts when BO was dried under RLD, BLD, RBLD, and WLD were 21.5%, 39.6%, 40.0%, and 51.6%, respectively, at a concentration of 5,000 ppm. All of these findings suggest that BO extracts dried under RLD, BLD, RBLD, and WLD have great potential as a cosmeceutical ingredient with good biological activity.
Lymph node micrometastasis in stage I and II rectal cancer
Hye-Yoon Choi(최혜윤),Kil Yeon Lee(이길연),Su Youn Lee(이수연),Youn Wha Kim(김윤화) 대한종양외과학회 2016 Korean Journal of Clinical Oncology Vol.12 No.2
Purpose: The aim of this study was to determine the rate of lymph node (LN) micrometastasis in patients with stage I and II rectal cancer. Methods: One hundred eighty patients with either stage I or II rectal carcinoma who underwent curative resection between 1995 and 2010 were included. Forty-eight patients received neoadjuvant chemoradiotherapy. Two sections from each LN were stained with hematoxylin and eosin (H&E) and with CK20 by immunohistochemistry (IHC), respectively. Results: A total of 2,257 LNs with a median of 12.5 LNs per patient were examined. For IHC staining, CK20-positive neoplastic cells were found in 4 of the 2,257 LNs (0.2%) from 3 of the 180 patients (1.7%), and all corresponding H&E re-stained sections confirmed that these neoplastic cells were present. Three of four neoplastic cells were micrometastasis, and one was macrometastasis. All occult neoplastic cells were found in 3 of the 85 patients (3.5%) with stage II disease. Conclusion: We observed a 3.5% rate of occult neoplastic cells in stage II rectal cancer. Interestingly, the results of IHC staining corresponded with those of H&E re-stained sections, suggesting that the examination of H&E stained section by a competent pathologist may replace IHC staining.
Ok Jeong Lee,Su-Jin Kim,박형두,이수연,Chi-Hwa Kim,Ah-Ra Ko,Yeon-Joo Yook,Su-Jin Lee,박성원,Se-Hwa Kim,Sung-Yoon Cho,Eun-Kyung Kwon,Sun Ju Han,진동규,Young Bae Sohn 대한소아청소년과학회 2012 Clinical and Experimental Pediatrics (CEP) Vol.55 No.3
Purpose: Mucopolysaccharidosis type II (MPS II or Hunter syndrome)is a rare lysosomal storage disorder caused by iduronate-2-sulfatase (IDS) deficiency. MPS II causes a wide phenotypic spectrum of symptoms ranging from mild to severe. IDS activity, which is measured in leukocyte pellets or fibroblasts, was reported to be related to clinical phenotype by Sukegawa-Hayasaka et al. Measurement of residual plasma IDS activity using a fluorometric assay is simpler than conventional measurements using skin fibroblasts or peripheral blood mononuclear cells. This is the first study to describe the relationship between plasma IDS activity and clinical phenotype of MPS II. Methods: We hypothesized that residual plasma IDS activity is related to clinical phenotype. We classified 43 Hunter syndrome patients as having attenuated or severe disease types based on clinical characteristics,especially intellectual and cognitive status. There were 27 patients with the severe type and 16 with the attenuated type. Plasma IDS activity was measured by a fluorometric enzyme assay using 4-methylumbelliferyl-α-iduronate 2-sulphate. Results: Plasma IDS activity in patients with the severe type was significantly lower than that in patients with the attenuated type (P=0.006). The optimal cut-off value of plasma IDS activity for distinguishing the severe type from the attenuated type was 0.63 nmol·4 hr-1·mL-1. This value had 88.2% sensitivity, 65.4% specificity, and an area under receiver-operator characteristics (ROC) curve of 0.768 (ROC curve analysis; P=0.003). Conclusion: These results show that the mild phenotype may be related to residual lysosomal enzyme activity.