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은행(銀杏) 종의(種衣)의 Phospholipase $C{\gamma}1$ 저해 활성 성분 (2)
이지숙,조유선,이현선,안종석,김진웅,Lee, Ji-Suk,Cho, You-Sun,Lee, Hyun-Sun,Ahn, Jong-Seog,Kim, Jin-Woong 한국생약학회 1999 생약학회지 Vol.30 No.3
Using the bioassay-guided fractionation and isolation technique, two $PLC{\gamma}1$ inhibitors were isolated from the sarcotestas of Ginkgo biloba (Ginkgoaceae). The structures of these compounds were identified as (3R)-(-)-8-hydroxy-3-(6'-pentadecenyl)3,4-dihydroisocoumarin (1) and 3-heptadecen-2-one (2) by UV, IR, MS, $^1H-NMR$, $^{13}C-NMR$ and $^1H-^{13}C\;COSY$. Isolate compounds 1 and 2 have not been reported previously from the sarcotestas of G. biloba and Ginkgoaceae, respectively. In addtion, these compounds showed significant $PLC{\gamma}1$ inhibitory effects with the $IC_{50}$ of the 9.7 (1) and $25.6\;{\mu}M\;(2)$.
Screening of Bioactive Materials from Freshwater Microalgae
이완석,최애란,안치용,오현철,안종석,오희목,Lee, Wan-Seok,Choi, Ae-Ran,Ahn, Chi-Yong,Oh, Hyun-Cheol,Ahn, Jong-Seog,Oh, Hee-Mock The Korean Society of Phycology 2004 ALGAE Vol.19 No.3
One hundred and fifty four micro algal strains, newly isolated from nationwide freshwaters in Korea, were screened for their anticancer, ant diabetic, and antibiotic activities. The micro algal strains were cultured with different nutritional conditions that were divided into 4 groups as follows; a normal Allen medium, nitrogen (N)-limited medium, phosphorus (P)-limited medium, and N and P-limited medium. Algal biomass was extracted with a mixture of acetone:H₂O (1:1, v:v) and the extracts were used for the screening of bioactive materials. Anticancer, ant diabetic, and antibiotic materials were screened by the methods of vaccinia Hl-related protein tyrosine phosphates (VHR DS-PTPase) inhibition, protein tyrosine phosphates 1B (PTP1B) inhibition, and paper disk. The inhibition activity of VHR DS-PTPase was observed in 18 strains, having a maximum 79% inhibition from Anabaena affinis and the inhibition activity of PTP1B was observed in 9 strains, having a maximum 97% from Sphaerocystis schroeteri. Microcystis aeruginosa incubated in an N and P-limited medium showed antibiotic activity in 8 species out of 13 pathogenic bacteria. As a whole, it seemed that the stressed condition such as N and/or P limitation increased the production of bioactive materials in micro algae.
Characterization of Sporulation-Specific Glucoamylase of Saccharomyces diastaticu
Eun Ju Kim(김은주),Jong Seog Ahn(안종석),Dae Ook Kang(강대욱) 한국생명과학회 2010 생명과학회지 Vol.20 No.5
효모 Saccharomyces diastaticus 는 세포 외로 분비되는 glucoamylase Ⅰ, Ⅱ, Ⅲ 동위효소 중 하나를 생산하여 전분을 가수분해하여 포도당을 생성할 수 있다. Glucoamylase Ⅰ, Ⅱ, Ⅲ는 STA1, STA2, STA3 유전자에 의해 각각 암호화된다. 효모 Saccharomyces 속이 포자가 형성되는 시기에 세포 내에서 특이적으로 발현된다고 알려진 glucoamylase(SGA)의 분자생물학적 및 생화학적 연구를 수행하기 위한 일환으로 S. diastaticus YIY 345 형질전환체의 배양 상등액으로부터 SGA 정제를 시도하였다. 황산암모늄 침전, DEAE-Sephadex A-50, CM-Sephadex C-50, Sephadex G-200 chromatography 등의 정제과정을 거쳐서 비특이 활성이 174배 증가된 0.22 ㎎의 순수한 SGA를 얻었다. HPLC와 SDS-PAGE 분석을 통해 이 효소는 63, 68 kDa의 단위체로 구성된 이합체임을 확인할 수 있었다. Con-A Sepharose 친화성 크로마토그피와 탈당쇄 효소를 처리한 결과로부터 SGA는 N-연결형 당쇄로 수식되었으며 단백질 부분은 59 kDa이었다. 정제한 SGA와 세포 외 분비성 glucoamylase의 효소학적 특성을 조사하고 비교한 결과 SGA의 최적 pH와 온도는 각각 5.5와 45℃로 나타났으며 세포 외 분비성 glucoamylase는 5.0과 50℃로 나타났다. SGA는 세포 외로 분비되는 glucoamylase에 비해 열처리 및 SDS에 대해 더 민감한 반응성을 나타내었다. The yeast strains of Saccharomyces diastaticus produce one of three isozymes of an extracellular glucoamylase Ⅰ, Ⅱ or Ⅲ, a type of exo-enzyme which can hydrolyse starch to generate glucose molecules from non-reducing ends. These enzymes are encoded by the STA1, STA2 and STA3 genes. Another gene, sporulation-specific glucoamylase (SGA), also exists in the genus Saccharomyces which is very homologous to the STA genes. The SGA has been known to be produced in the cytosol during sporulation. However, we hypothesized that the SGA is capable of being secreted to the extracellular region because of about 20 hydrophobic amino acid residues at the N-terminus which can function as a signal peptide. We expressed the cloned SGA gene in S. diastaticus YIY345. In order to compare the biochemical properties of the extracellular glucoamylase and the SGA, the SGA was purified from the culture supernatant through ammonium sulfate precipitation, DEAE-Sephadex A-50, CM-Sephadex C-50 and Sephadex G-200 chromatography. The molecular weight of the intact SGA was estimated to be about 130 kDa by gel filtration chromatography with high performance liquid chromatography (HPLC) column. Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed it was composed of two heterogeneous subunits, 63 kDa and 68 kDa. The deglycosylation of the SGA generated a new 59 kDa band on the SDS-PAGE analysis, indicating that two subunits are glycosylated but the extent of glycosylation is different between them. The optimum pH and temperature of the SGA were 5.5 and 45℃, respectively, whereas those for the extracellular glucoamylase were 5.0 and 50℃. The SGA were more sensitive to heat and SDS than the extracellular glucoamylase.
쿠마린에 의한 RAW 264.7 세포주의 Nitric Oxide 생성 저해활성
노태철,최희철,김보연,김영호,안종석,김영국,이현선,Rho, Tae-Cheol,Choi, Hee-Cheol,Kim, Bo-Yeon,Kim, Young-Ho,Ahn, Jong-Seog,Kim, Young-Kook,Lee, Hyun-Sun 한국생약학회 1999 생약학회지 Vol.30 No.4
During the screening for inhibitors on nitric oxide (NO) production in LPS-activated murine macrophage, RAW 264.7 cells, two coumarins were isolated from chloroform extract of Ponciri Fructus. They were identified as imperatorin (10), phellopterin (11) on the basis of spectroscopic methods. The $IC_{50}$ values for NO formatiom were about $5.4\;{\um}M$ and $35.0\;{\um}M$, respectively, and then eleven coumarins were tested for the inhibitory effects on NO production in activated macrophages. All the test coumarins inhibited NO production in concentration-dependent manner and furanocoumarins (6-11) showed much more potent inhibitory effect than simple coumarins. Among the compounds examined xanthotoxin (8) was the most potent inhibitors of NO production $(IC_{50}=1.4\;{\mu}M)$. Analysis of the structure-activity relationship among these coumarins led to the conclusion that the substitution of C-5 position in furanocoumarins reduces greatly their inhibitory potency although the substitution of C-8 does not almost affect it.
갯기름나물의 쿠마린에 의한 RAW 264.7 세포주의 Nitric Oxide 생성 저해활성
최희철,노태철,김보연,고학룡,오원근,성창근,민태익,안종석,이현선,Choi, Hee-Cheol,Rho, Tae-Cheol,Kim, Bo-Yeon,Ko, Hack-Ryong,Oh, Won-Keun,Seong, Chang-Keun,Mheen, Tae-Ick,Ahn, Jong-Seog,Lee, Hyun-Sun 한국생약학회 1999 생약학회지 Vol.30 No.2
During the screening for inhibitors of nitric oxide production in LPS-activated macrophage, RAW 264.7 cells. Five coumarins were isolated from chloroform extract of the root of Peucedanum japonicum. They were identified as praeruptorin A (1), xanthotoxin (2), psoralen (3), isopimpinellin (4), bergapten (5) on the basis of spectroscopic methods. The $IC_{50}$ values for nitrite production by activated macrophages were about $1.5\;{\mu}g/ml$ (1), $0.3\;{\mu}g/ml$ (2), $1.0\;{\mu}g/ml$ (3), $25\;{\mu}g/ml$ (4), $25\;{\mu}g/ml$ (5), respectively. However, the inducible nitric oxide synthase (iNOS) was not inhibited by treatment with these compounds. Their inhibitory effect on nitric oxide production was resulted from the supperssion of iNOS expression.
백자인 추출물에 의한 pancreatic lipase의 저해 효과
김민수,김보연,박찬선,윤병대,안순철,오원근,안종석,Kim Min-Soo,Kim Bo-Yeon,Park Chan-Sun,Yoon Byung-Dae,Ahn Soon-Cheol,Oh Won-Keun,Ahn Jong-Seog 한국생명과학회 2006 생명과학회지 Vol.16 No.2
Pancreatic lipase의 활성을 저해하는 물질을 탐색하고자 국내의 식용 또는 약용식물의 추출물을 대상으로 lipase 활성에 대한 저해능력이 있는 식물추출물은 탐색한 결과 대복피, 계혈등, 오배자 및 백자인의 식물추출물을 선정하였다. Pancreatic lipase에 대한 저해활성이 우수한 백자인으로부터 클로로포름 용매추출, 실리카겔 컬럼크로마토그라피, 세파덱스 LH-20 컬럼크로마토그라피와 고속액체크로마토그라피를 실시하여 pancreatic lipase에 대한 저해활성 물질로 TF-1, TF-2, TF-3를 분리하였다. 이들을 대상으로 porcine pancreatic lipase에 대한 저해효과를 측정한 결과, 활성물질 TF-1, TF-2, TF-3 및 orlistat의 porcine pancreatic lipase에 대한 $IC_{50}$ 값은 각각 44.7, 98.7, 46.1 및 $27.6{\mu}g/ml$인 것으로 나타났다. 활성물질 TF-2와 orlistat의 경우에는 $10{\mu}g/ml$의 농도에서 NIH-3T3 L1의 지방세포로의 분화에도 억제효과가 있음을 입증하였다. The possible presence of inhibitors of pancreatic lipase (tricaylglycerol acylhydrolase EC 3.1.1.3) was screened from Korean traditional edible or medicinal herbs. Among tested herbs, Arecae pericarpium, Mucunae Caulis, Rhus javanica, Thujae orientalis were shown to have strong inhibitory effect against pancreatic lipase. Thujae orientalis was finally selected as a candidate for pancreatic lipase inhibitor. The extract of Thujae orientalis was showed selective inhibition on porcine pancreatic lipase activity. Active inhibitors, TF-1, TF-2, TF-3, were purified from an extract of Thujae orientalis, using chloroform extraction, followed by successive chromatography in silica gel and LH-20 and high performance liquid chromatography (HPLC). The $IC_{50}$ values of TF-1, TF-2, TF-3 and orlistat were 44.7, 98.7, 46.1 and $27.6{\mu}g/ml$, respectively. And also the TF-2 and orlistat were shown to be inhibitory effect on the differentiation of preadipocyte NIH-3T3 L1 cells at a concentration of $10{\mu}g/ml$.
오원근,강대욱,박찬선,안순철,고학룡,김보연,민태익,안종석,이현선,Oh, Won-Keun,Kang, Dae-Ook,Park, Chan-Sun,Ahn, Soon-Cheol,Ko, Hack-Ryong,Kim, Bo-Yeon,Mheen, Tae-Ik,Ahn, Jong-Seog,Lee, Hyun-Sun 한국생약학회 1997 생약학회지 Vol.28 No.1
The total MeOH extracts of 149 herbal medicines were screened far angiotensin ll receptor antagonistic activity using rat liver membrane fraction. Four medicinal plants- Atractylodes japonica, Evodia officinalis, Scutellaria baicalensis, Machilus thunbergii showed strong angiotensin ll antagonistic activity.