녹용의 약효 성분에 관한 연구 ( 3 ) 녹용 및 Pantocrin 의 지방산 조성에 관하여

김영은,이승기,이명희,신승언 ( Young Eun Kim,Seung Ki Lee,Myoung Hee Lee,Seung Uon Shin ) 생화학분자생물학회 1976 BMB Reports Vol.9 No.4

The lipid soluble fraction of antler velvet layer (Cervus nippon taiouanus) was extracted and compared to that of pantocrin (ethanol preparation of antler, commericially available). The lipid soluble components(801.5㎎ from antler velvet layer and 979.1㎎/150㎖ from pantocrin) were fractionated into neutral lipids, glycolipids, phosphclipids, gangliosides, and nonlipid components (lipoprotein and peptide) on Sephadex G-25 column followed by silicic acid column chromatography. The percentage composition of the lipid soluble fraction was neutral lipid 65.3%, glycolipid 5.2%, phospholipid 12.9%, ganglioside 6.1 %, nolipid 2.9 %. The neutral lipids were fractionated into squalene, sterolester, triglyceride, free fatty acid, sterol, diglyceride, monoglyceride on boric acid impregnated silicic acid column. The fractions collected were determined by thin layer chromatography. The compositions of the free fatty acid fractions were determined by gas liquid chromatography. The sterolester, triglyceride fractions were hydrolyzed to give free fatty acids with 10% KOH solution, refluxed at 80-85℃ for four hours, and carried out gas liquid chromatography. The free fatty acids in antler velvet layer were found to be nine kinds of saturated fatty acids, nine kinds of unsaturated fatty acids, two branched fatty acids and six unknown components. The main components of antler free fatty sacid were oleic (21.7%), linolenic(31.2%), palmitic(19.6%), stearic(12.1%), palmitoleic(10.2%), and those make 94.4% of antler free fatty acids. In pantocrin free fatty acid fraction, seven kinds of saturated fatty acids, four kinds of unsaturated fatty acids, one branched fatty acid and three unknown compopents were detected. The main components of pantocrin free fatty, acid fraction wereoleic(45%), palmitic(19.5%), lauric(10.9%), stearic(6.6%), palmitoleic(5.6%), myristic(3.4%), capric(3.1%), and those make 94.1% of pantocrin free tfatty acids. The ester fatty acids of triglyceride fraction were found as follows: In antler triglyceride faction, nine kinds of saturated fatty acids, three kinds of unsaturated fatty acids, three branched fatty acids and ten unknown components were detected. In pantocrin triglyceride fraction, it was found to contain five kinds of saturated fatty acids, two kinds of unsaturated fatty acids, one branched fatty acid and ten unknown components. after saponification of triglyceride fraction, two new fatty acids (pentadecylic, oleic) in antler neutral lipid and two new fatty acids (lauric, dodecenoic) in pantocrin neutral lipid were detected. In sterol ester fraction, there were six saturated fatty acids, three unsaturated fatty acids, one branched fatty acid and eight unknown components in antler neutral lipid, and five saturated fatty acid, three unsaturated fatty acids and five unknown components in pantocrin neutral lipid. After saponification of sterol ester fractions, new fatty acids were determined as dodecenoic, lauric, anteiso-pentadecylic, isopalmitic, heptadecanoic in antler sterol ester fraction and dodecenoic, palmitic, heptadecanoic, heptadecenoic in pantccrin sterol ester fraction.

녹용의 약효 성분에 관한 연구(IV) 녹용 벨?殼?의 프로스타그란딘의 검출

김영은,이승기,이명희,Kim, Young-Eun,Lee, Seung-Ki,Lee, Myoung-Hee 생화학분자생물학회 1977 한국생화학회지 Vol.10 No.1

녹용의 지용성 성분에 관한 생화학적 연구의 일환으로 대만산 꽃사슴(cervus nippou taiouanus)의 녹용을 절단한 후 동결하여 벨?殼?과 해면상 골조직으로 분리하였다. 벨?殼?을 절편으로 만든 후 0.9% saline 용액중에서 Virtis homogenizer로 파쇄하여 Jouvenaz 의 방법에 따라 prostaglandin을 추출하였다. Anderson, Samuelsson 등의 방법에 준하여 thin layer chromatography를 행하여 $PGE_2$, 15-epi-$PGE_1$, $PGE_{1{\alpha}}$ 및 $PGE_{1{\beta}}$로 추정되는 물질을 검출하였으며 preparative T.L.C. 를 행하여 분리한 후 Samuelsson의 방법에 따라 U.V. spectroscopy를 행하여 $PGE_2$, 15-epi-$PGE_1$으로 추정되는 분획에서는 알카리 처리 후에 278nm에서 U.V. 흡수를 나타냈으며 한편 $PGE_{1{\alpha}}$, $ PGE_{1{\beta}}$로 추정되는 분획에서는 알칼리 처리 전후에 U.V. 흡수가 없었으므로 T.L.C의 결과와 U.V. spectroscopy의 결과는 상호간에 일치됨을 알 수 있었다. 또한 Fishbein 등의 방법에 의한 gas-liquid chromatography를 행하여 $PGE_2$ standard peak와 일치하는 peak를 얻어 $PGE_2$의 존재를 확인하였으며 retention time이 $PGE_{1{\alpha}}$ 및 $PGE_{1{\beta}}$와 일치되는 peak를 얻어 세방법에서 공히 $PGE_2$, 15-epi-$PGE_1$, $PGE_{1{\alpha}}$, $PGE_{1{\beta}}$의 존재를 검출하였다. The fresh antler was cut, frozen and mechanically separated into spongy bone layer and velvet layer. The prostaglandin-like components were extracted from antler velvet layer and qualitatively analyzed by three different methods: thin layer chromatography, ultra-violet spectroscopy after conversion to PGB series by alkaline treatment, and gas liquid chromatography. The prostaglandins detected were $PGE_2$, 15-epi-$PGE_1$, $PGE_{1{\alpha}}$, $PGE_{1{\beta}}$. The experiments indicated that the petroleum ether fraction contained mostly PGE series and that the ethylether fraction contained PGF series. One major prostaglandin-like components of antler velvet layer was found to be the main compound in the petroleum ether extract. The structure of this compound is still unknown but the gas liquid chromatography data suggest that it is a hydroxy fatty acid.

동물 경조직 단백성분의 조성과 생리기능에 관한 연구 녹각의 경단백질에 대하여

김영은,이승기,윤웅찬,Kim, Young-Eun,Lee, Seung-Ki,Yoon, Ung-Chan 생화학분자생물학회 1973 한국생화학회지 Vol.6 No.1

동물 경조직 단백성분의 연구의 일환으로 녹각 및 녹용의 생화학적 기능을 나타내는 성분을 밝히기 위하여 녹각 분말을 0.5M 초산으로 탈칼슘을 한 후 5M 요소로 단백질부분을 추출하였다. 추출한 녹각단액의 아미노산 구성은 16종류에 달했으며 이중 glycine은 24.51%; proline 11.09%; glutamic acid는 10.26%가 되어 검출된 전 아미노산의 45.86%를 차지하고 있음이 밝혀졌다. 박충크로마토그라피로 glucose와 galactose 및 hexoamine의 존재를 확인하였고 정량분석 결과 총 hexose의 양은 추출단백의 0.74%이고, 총 hexoamine의 양은 0.87%였다. 본보에서 보고한 바와 같은 아미노산의 구성비율로 미루어 녹각단백은 일종의 collagen으로 추정하였다. 그러나 녹각단백중에 존재하는 아미노산 구성중 특기할 점은 cystine이 존재하는 반면 valine이 검출되지 않았다는 점이다. emission spectroscopy로 검출된 무기원소의 종류는 13종이며 회화잔사의 함량은 녹각무게의 55.6% (w/w)이며 이 회화잔사중 Ca은 회화잔사의 36.79%이고 Mg은 1.42%를 차지하고 있음을 청량분석의 결과 규명하였다. With an attempt to elucidate the biologically functioning groups of deer horn, we obtained, in this investigation, a protein fraction after extraction of the pulverized deer horn with 5M-urea followed by decalcification with acetic acid. The amino acid composition of deer horn extracted came up to 16 kinds. And analysis of the protein extracted with 5M-urea gave values, as per cent, glycine, 24.51%; proline, 11.09%; glutamic acid, 10.26%, which come up to 46.86% of the total amino acids initially present. The existence of glucose and galactose is confirmed with the results of thin-layer chromatography being carried out many times. And the total amounts of hexose and hexosamine are accounting for 0.74% and 0.87% for each. The present communication described as above has affirmed the protein from deer horn to be a sort of collagen. However, it can be focused on the significance of the existence of cystine and the absence of valine in deer horn. The contents of inorganic elements detected with emission spectroscophy are listed as following: total inorganic elements are detected to be 13 kinds. And the ash-contents of deer horns are 55.6% (w/w). The quantitative analysis for the ash shows that it contains 36.79% of Ca and 1.42% of Mg against ash contents by weight per weight.

녹용의 약효성분에 관한 연구 ( 2 ) 녹용의 Acid Mucopolysaccharide 성분에 관하여

김영은,이승기,유희자 ( Young Eun Kim,Seung Ki Lee,Hee Ja Yoo ) 생화학분자생물학회 1976 BMB Reports Vol.9 No.3

A fresh antler from Cervus nippon taiouanus was cut and frozen. The constituents were divided into water soluble and lipid soluble fractions for elucidation of the chemical composition and structure of the antler. For the water soluble fraction of the antler we reported the data in this Journal(Kim et a1.1975). The data of qualitative and quantitative analysis of neutral sugar, hexosamine, uronic acid, hydroxyproline, ash, ester sulfate, sialic acid and total nitrogen were reported. In the present paper, acid mucopolysaccharides were isolated from antler velvet layer, antler spongybone layer and whale nasal cartilage by the extraction procedure of Mathews and Kubota. Acid mucopolysaccharides were extracted by digestion with pepsin, papain followed by hydrolysis with alkaline solution. The yield of acid mucopolysaccharides from antler velvet layer was approximately 0.16% to the defatted dried weight of antler velvet layer. Fractionation was performed by ECTEOLA-cellulose column chromatography, developing with 0.05∼2.5M NaCl and HCl (equimolarly mixed) as given by Ringertz et al. The elution profile was divided into five fractions, namely Fr. I(0.05M-Cl`), Fr. II (0.05M-Cl`), Fr. III (0.1M-Cl`), Fr. IV (0.2M-Cl`) and Fr. V (0.5M-Cl`) from antler velvet layer, and four fractions, Fr. I`(0.5M-Cl`), Fr. II` (0.7M-Cl`), Fr. III` (1.1M-Cl`) and Fr. IV` (2.5M-Cl`) from whale nasal cartilage. The nature of each fraction was examined by celllulose acetate membrane electrophoresis and infrared spectroscopy. Fraction I of antler velvet layer (eluted with 0.05 M-Cl`) showed identical electrophoretic mobility and absorbance pattern for infrared spectroscopy to hyaluronic acid standard. Fr. V from antler velvet layer and Fr. I` from whale nasal cartilage (eluted with 0.5M-Cl`) showed the same electrophoretic mobility and gave identical spectra to chondroitin sulfate A standard.

녹용의 약효 성분에 관한 연구 ( 1 ) 녹용 , 녹각 , 고래 코 연골 , 상어 척수 연골의 화학조성에 관하여

김영은,이승기,윤웅찬,김정숙 ( Young Eun Kim,Seung Ki Lee,Ung Chan Yoon,Jung Sook Kim ) 생화학분자생물학회 1975 BMB Reports Vol.8 No.2

In this study, an attempt has been made to investigate the differences between the chemical components of antler (Corpus Cervi Parvum) and old antler (Cornus Cervi) . Antler was cut from Cervus nippon taiouanus (3 years and 8 months old) and directly fronzen to storage in solid carbon dioxide. The antler was separated into velvet layer ancf spongy bone layer and cut into small pieces. After dehydration and removal of lipids from the antler with acetone, chloroform-methanol (1 : 1) solvent system, qualitative and quantitative analyses were carried out to compare their chemical components in tissue level with those of old antler spongybone layer, shark backbone cartilage and whale nasal cartilage. Quantitative analyses of hexose, pentose, hexosamine, uronic acid, sialic acid, ester-sulfate, hydroxyproline, total nitrogen and inorganic contents were carried out respectively for antler velvet layer, antler spongybone layer, old antler spongybone layer, shark backbone cartilage and whale nasal cartilage. And their relative amounts of the chemical components to the corresponding dry weight of samples were listed in Table XII. The neutral sugar compositions of samples were identified by gas-liquid chromatography. Antler velvet layer, old antler spongybone layer and whale nasal cartilage contained same kinds of neutral sugars, such as glucose, galactose, mannose, fucose, deoxyribose, ribose, arabinose and xylose. But antler spongybone layer and shark backbone cartilage did not show any sign of existence of fucose though they contained the same kinds of neutral sugars as the above. The gas liquid chromatography of aminosugars was also carried out. The results said that all five animal tissues contained both glucosamine and galactosamine. From the results of analytical data, the presence of mucopolysaccharides in antler and old antler was revealed. And the contents of hydroxyproline and total nitrogen of samples suggested that the antler-protein was mainly composed of collagen. The relative amounts of hydroxyproline to corresponding dry weight of samples showed that antler velvet layer exhibited the highest amounts (9.09%) and shark backbone cartilage (3.50%) exhibited the lowest amounts of hydroxyproline. The mucopolysaccharide of whale nasal cartilage was known mainly as chondroitin sulfate A and that of shark backbone cartilage mainly as chondroitin sulfate C. A further study on the mucopolysaccharide-protein complex of antler and of old antler is being carried out and will be presented in the next report.

동물 경조직 단백성분의 조성과 생리기능에 관한 연구 녹각의 경단백질에 대하여

김영은,이승기,윤웅찬 ( Young Eun Kim,Seung Ki Lee,Ung Chan Yoon ) 생화학분자생물학회 1973 BMB Reports Vol.6 No.1

With an attempt to elucidate the biologically functioning groups of deer horn, we obtained, in this investigation, a protein fraction after extraction of the pulverized deer horn with 5M-urea followed by decalcification with acetic acid. The amino acid composition of deer horn extracted came up to 16 kinds. And analysis of the protein extracted with 5M-urea gave values, as per cent, glycine, 24.51 % ; proline, 11.09% ; glutamic acid, 10.26%, which come up to 46. 86% of the; total amino acids initially present. The existence of glucose and galactose is confirmed with the results of thin-layer chromatography being carried out many times. And the total amounts o_f hexose and hexosamine are accounting for 0.74% and 0.87% for each. The present communication described as above has affirmed the protein from deer horn to be a sort of collagen. However, it can be focused on the significance of the existence of cystine and the absence of valine in deer horn. The contents of inorganic elements detected with emission spectroscophy are listed as following: total inorganic elements are detected to be 13 kinds. And the ash-contents of deer horns are 55.6% (w/w). The quantitative analysis for the ash shows that it contains 36.79 of Ca and 1.42% of Mg against ash contents by weight per weight.

녹용의 약효 성분에 관한 연구 ( Ⅳ ) 녹용 벨벳층의 프로스타그란딘의 검출

김영은,이승기,이명희 ( Young Eun Kim,Seung Ki Lee,Myoung Hee Lee ) 생화학분자생물학회 1977 BMB Reports Vol.10 No.1

The fresh antler was cut, frozen and mechanically separated into spongy bone layer and velvet layer. The prostaglandin-like components were extracted from antler velvet layer and qualitatively analyzed by three different methods: thin layer chromatography, ultra-violet spectroscopy after conversion to PGB series by alkaline treatment, and gas liquid chromatography. The prostaglandins detected were PGE₂, 15-epi-PGE₁, PGF_(1α), PGF_(1β). The experiments indicated that the petroleum ether fraction contained mostly PGE series and that the ethylether fraction contained PGF series. One major prostaglandin-like components of antler velvet layer was found to be the main compound in the petroleum ether extract. The structure of this compound is still unknown but the gas liquid chromatography data suggest that it is a hydroxy fatty acid.

동물 경조직 단백 성분의 조성과 생리기능에 관한 연구 - 우각 수우각 서각에서 분리한 겸 단백의 조성에 대하여

이승기,김영은,Lee, Seung-Ki,Kim, Young-Eun 생화학분자생물학회 1974 한국생화학회지 Vol.7 No.2

동물 경조직 단백에 관한 연구의 일환으로 3종의 동물의 각질(우각, 수우각, 서각)을 각각 채취하여 Gillespie 등의 방법에 의하여 0.1 M-thioglycolate로 $400^{\circ}C$에서 keratin을 추출한 후 S-carboxymethyl (SCMK) 유도체를 합성하였고 $\frac{1}{10}$ N-HCl 로 등전점 (pI 4.7~5.05)의 범위에서 침전된 단백을 각각 분리하였다. 추출된 경단백의 수득량은 건조 중량으로 16~30%이었다. 추출된 keratin의 아미노산 조성은 3종 공히 18종이었으며, 그 조성비의 차이는 별로 없었으나 단 우각에서 추출된 단백질의 Lysine 함량은 2.67%로 수우각 5.57%, 서각 4.99%에 비하여 현저한 차이가 있었으며, 우각의 S-carboxymethyl cysteine의 함량은 9.41%로 수우각 6.06%, 서각 5.96%에 비해 크므로 우각 keratin의 disulfide bridge에 의한 cross-linking 정도가 수우각, 서각에 비해 크다는 것 을 나타내고 있다. 추출 단백의 monosaccharide, sialic acid, hexosamine, 및 uronic acid의 조성을 분석한 결과 이들 총 함량을 표시하면 추출 단백질의 건조중량에 대하여 우각은 1.38%, 수우각 0.47%, 서각 1.04%이었다. Sialic acid의 함량은 추출 단백의 건조중량에 대하여 gm 당 우각 $1.26{\mu}mol$, 수우각 $0.29{\mu}mol$, 서각 $0.96{\mu}mol$로 나타났다. Hexosamine 분석에서 3종 공히 galactosamine은 검출 되지 않고 glucosamine만이 검출?瑛만? 또한 uronic acid 분석결과 glucuronic acid는 3종 공히 검출되지 않았고 galacturonic acid만이 검출되었다. 또한 중성당의 조성은 우각 및 수우각에는 glucose, galactose, xylose, fucose가 서각에는 glucose, mannose, ribose가 각각 검출되었다. We have previously reported that the abstinence syndrom of narcotic addicts was relieved by the protein extracts of animal hard tissue (from Manis pentadactyla). The protein extracts of the hard tissue were found to be a sort of keratin. Keratins were extracted from three species of animal horns (bovine horn, water buffalo horn and rhinoceros horn) with 0.1 M-Thioglycolate at $40^{\circ}C$ and S-carboxymethyl derivatives (SCMK) of the keratin were synthesized by the method of Gillespie and his co-workers. The yields of the keratin derivatives were in the range of 16% to 30% of the dry weight of the horns. In an attempt to elucidate and compare the compositions of the keratin derivatives, the analysis for amino acids, monosaccharides. hexosamines, uronic acids, sialic acids among bovine horn, water buffalo horn, and rhinoceros horn, were performed. Eighteen kinds of amino acids were found in the three keratin derivatives and there were no differences in the composition among them, with the exception of lysine contents of bovine horn protein extracts (2.67%), which was distinctively lower than those of water buffalo horn protein extracts (5.57%) and those of rhinoceros horn protein extracts (4.99%). The amounts of carboxymethyl cysteine of bovine horn are twice of those of water buffalo horn and those of rhinoceros horn. In view of these results, it is presumed that the crosslinking rate of bovine horn keratin by cystine disulfide bridge is higher than those of water buffalo horn and Rhinoceros horn. The conjugated components of the keratin derivatives, such as monosaccharides, sialic acids, hexosamines, and uronic acids were totally contained 1.38% in bovine horn keratin, 0.47% in water buffalo horn keratin and 1.04% in rhinoceros horn keratin. There are some similarities among the keratin derivatives of the three species, such as these keratins do not contain galactosamine but glucosamine, and also the existance of glucuronic acid in these keratins is not detected but galacturonic acid. The sugar moieties of these proteins consisted of glucose, galactose, xylose, fucose in bovine horn protein extracts; glucose, galactose, xylose, fucose in water buffalo horn protein extracts; glucose, mannose, ribose in rhinoceros horn protein extracts.

동물 경조직 단백 성분의 조성과 생리기능에 관한 연구 우각 수우각 서각에서 분리한 경단백의 조성에 대하여

이승기,김영은 ( Seung Ki Lee,Young Eun Kim ) 생화학분자생물학회 1974 BMB Reports Vol.7 No.2

We have previously reported that the abstinence syndrom of narcotic addicts was relieved by the protein extracts of animal hard tissue (from Manis pentadactyla). The protein extracts of the hard tissue were found to be a sort of keratin. Keratins were extracted from three species of animal horns (bovine horn, water buffalo horn and rhinoceros horn) with 0.1 M-Thioglycolate at 40℃ and S-carboxymethyl derivatives (SCMK) of the keratin were synthesized by the method of Gillespie and his co-workers. The yields of the keratin derivatives were in the range of 16% to 30% of the dryweight of the horns. In an attempt to elucidate and compare the compositions of the keratin derivatives, the analysis for amino acids, monosaccharides, hexosamines, uronic acids, sialic acids among bovine horn, water buffalo horn, and rhinoceros horn, were performed. Eighteen kinds of amino acids were found in the three keratin derivatives and there were no differences in the composition among them, with the exception of lysine contents of bovine horn protein extracts(2.67%), which was distinctively lower than those of water buffalo horn protein extracts(5.57%) and those of rhinoceros horn protein extracts (4.99%). The amounts of carboxymethyl cysteine of bovine horn are twice of those of water buffalo horn and those of rhinoceros horn. In view of these results, it is presumed that the crosslinking rate of bovine horn keratin by cystine disulfide bridge is higher than those of water buffalo horn and Rhinoceros horn. The conjugated components of the keratin derivatives, such as monosaccharides, sialic acids, hexosamines, and uronic acids were totally contained 1.38% in bovine horn keratin, 0.47% in water buffalo horn keratin and 1.04 in rhinoceros horn keratin. There are some similarities among the keratin derivatives of the three species, such as these keratins do not contain galactosamine but glucosamine, and also the existance of glucuronic acid in these keratins is not detected but galacturonic acid. The sugar moieties of these proteins consisted of glucose, galactose, xylose, fucose in bovine horn protein extracts; glucose, galactose, xylose, fucose in water buffalo horn protein extracts; glucose, galactose, mannose, ribose in rhinoceros horn protein extracts.

xEV차량 구동모터 기반 차량 긴급 핸들링 보조 로직(E-EHA) 개발

박재일(Jae Il Park),김승기(Seung Ki Kim),황성욱(Sung Wook Hwang),이상호(Sang Ho Lee),김상준(Sang Joon Kim),유성훈(Sung Hoon Yu),김영은(Young-eun Kim),김우균(Woo Kyun Kim) 한국자동차공학회 2021 한국자동차공학회 부문종합 학술대회 Vol.2021 No.6