참당귀(Angelica gigas Nakai) 뿌리의 면역증강활성 성분

안경섭,심웅섭,김환묵,한상배,김익환,Ahn, Kyung-Seop,Sim, Woong-Seop,Kim, Hwan-Mook,Han, Sang-Bae,Kim, Ik-Hwan 한국생약학회 1996 생약학회지 Vol.27 No.3

A potent immuno-stimulating activity was detected from the watersoluble and ethanol-precipitated crude extract (AG-1) of the root of Angelica gigas Nakai. The crude extract was fractionated into two fractions, an acidic AG-2 and a neutral AG-3 fraction by DEAE-cellulose adsorption. The two fractions contained polysaccharides of which M.W. were 10 Kdal and >2,000 Kdal respectively, proteins, and various inorganic components. The immunostimulating activities of two fractions were not reduced by proteinase K, acid or alkali treatment. The polysaccharides obtained from the root of A. gigas were mainly composed of arabinose, galactose, and galacturonic acid. These results indicated that immuno- stimulating components of the root of A. gigas was a kind of pectic polysaccharides or arabinogalactans.

발아 중인 옥수수에서 리보조옴 단백질의 인산화반응에 미치는 GA3 의 효과

안경섭(Kyung Seop Ahn),음진성(Jin Seong Eum),심웅섭(Woong Seop Sim) 한국식물학회 1990 Journal of Plant Biology Vol.33 No.1

In order to study the effect of GA_3 on the phosphorylation of ribosomal proteins during germination in Zea mays, ribosomal proteins were labelled with ^32P, extracted, electrophoresed and autoradiographed. There are five phosphorylated ribosomal proteins. One of these is in 40S subunit and has molecular weight of 33,000 daltons. Others are in 60S subunit and have molecular weights of 37,000, 16,000, 15,200 and 13,500, respectively. Phosphorylation of ribosomal proteins was increased maximum 47.7% in shoots of Zea mays treated with GA_3.

Metabolomic approach of Korean Hypericum species using UPLC–QTOF-MS

Hyuk-Hwan Song(송혁환),Hyung Won Ryu(류형원),Ok Kyoung Kwon(권옥경),Kyung Seop Ahn(안경섭),Kyeong Yel Oh(오경열),Jun Young Kim(김준영),Sei-Ryang Oh(오세량) 한국약용작물학회 2013 한국약용작물학회 학술대회논문집 Vol.2013 No.2

면역증강성 다당 생산을 위한 참당귀 세포배양의 최적조건

안경섭,서원택,심웅섭,김익환 한국산업미생물학회 1998 한국미생물·생명공학회지 Vol.26 No.2

참당귀 세포(Angelica gigas H4)를 현탁배양하여 면역증강성 다당을 생산하기 위한 최적배지 설계 및 생산조건 최적화 연구를 수행하였다. 우선 세포의 성장에 가장 좋은 배지로서 Schenk and Hildebrandt(SH) 배지를 선택하여 배양한 결과 25℃, 암배양조건에서 각각 15.8 g DCW/ℓ와 0.85 g/ℓ의 세포 및 다당을 얻을 수 있었다. 한편 다당 생산용 배지의 개발을 위하여 여러 가지 식물세포배양용 배지중에서 다당생산능이 가장 높은 배지로서 Gamborg B5 배지를 선정하였으며 B5 배지를 변형하여 면역증강성 다당생산을 위한 최적배지(PPM)를 개발하였다. 그리고 최적배지를 이용한 당귀 세포증식과 다당 생산을 위한 최적 pH는 6.0-6.5, 배양온도는 20℃, 그리고 광조건보다는 암조건이 좋은 것으로 밝혀졌다. 상기 최적조건에서의 최대 세포증식과 다당생산을 각각 14.8 g/ℓ, 1.5 g/ℓ이었다. An Immunostimulating polysaccharide was produced from the suspension culture of Angelica gigas H4 plant cells. In order to enhance the polysaccharide production by the A. gigas cell culture, medium composition and physical conditions were optimized. Schenk and Hildebrandt (SH) medium was selected as an optimal basal medium for the growth of A. gigas. The maximum cell and polysaccharide concentration obtained in SH medium were 15.8 g DCW/ℓ and 0.85 g polysaccharide/ℓ, respectively, at 25℃ under dark condition. For the enhanced polysaccharide production, a polysaccharide production medium (PPM) was established by modifying Gamborg B5 medium with optimized carbon sources, growth regulators, organic and inorganic elements. Optimal initial pH and temperature were 6.0-6.6 and 20℃, respectively, and the dark condition was better than the light condition. The maximum polysaccharide concentration of 1.5 g/ℓ could be obtained through the optimization of the medium composition and physical conditions.

효소공정에 의한 트립토판 생산

이인영,안경섭,김익환,이선복 한국산업미생물학회 1992 한국미생물·생명공학회지 Vol.20 No.1

Tryptophanase를 이용하여 트립토판 합성시 인돌의 효소활성 저해를 억제하기 위하여 유가식 조업, 유기용매 이상계의 사용, 그리고 cyclodextrin의 첨가 등에 대하여 연구하였다. 효소 농도가 0.5㎎/㎖일 때 인돌 농도 0.4mM 부근에서 트립토판 생성이 가장 빨랐으며 그 이상에서는 효소활성이 심한 저해를 받았다. 초기 인돌 농도가 20mM일 때는 27시간 반응 후 인돌의 전환율이 20%인데 비하여 반응기내 인돌 농도를 5mM 이하로 유가식 조업을 하였을 때 전환율이 80%로 향상되었다. 트립토판 합성에 적합한 유기용매로 cyclohexane을 선정하였다. Cyclohexane/water 이상계에서 인돌은 주로 유기용매 층에 분배되었으며 계면활성제 Aliquat 336을 첨가하였을 때 트립토판은 pH가 올라감에 따라 cyclohexane 층으로 보다 많이 분배되었다. 그러나 유기용매 이상계에서 트립토판 합성은 원활하지 않았는데, 인돌이 대부분 유기용매 층에 존재함으로 인하여 그 속도가 늦어지는 것으로 생각된다. 한편 수용액에서 cyclodextrin을 첨가하여 트립토판 생성을 관찰한 결과 α-cyclodextrin은 트립토판 합성에 큰 영향을 미치지 않은 반면 β-cyclodextrin을 첨가하였을 때는 트립토판 합성속도가 현저히 증가하였다. Enzymatic synthesis of L-tryptophan(Trp) using E. coli tryptophanase has been investigated. In order to reduce the substrate inhibition by indole and to increase the product yield of L-tryptophan three different approaches have been made in this work. First, indole was intermittently fed to the reaction mixture in order to control the indole concentration at lower level. When 15mM of indole was used as a total amount of substrate, conversion yield of 80% has been obtained with intermittent feeding while only 20% of indole was converted into L-tryptophan by conventional batch operation. The second method employed in this work was the use of cyclohexane-phosphate buffer organic two-phase system. In this system, indole was mainly partitioned into the organic-solvent phase and therefore substrate inhibition was expected to be reduced. L-Tryptophan production in organic two-phase system was, however, unexpectedly lower than that obtained in aqueous buffer solution. As a third method cyclodextrins have been added to the aqueous reaction mixture. It was found that the addition of β-cyclodextrin enhanced the tryptophan synthesis noticeably while α-cyclodextrin showed little effect on tryptophan production.

유기용매 이상계에서의 Thermolysin에 의한 아스파탐 전구체 생산

이인영,안경섭,이선복 한국산업미생물학회 1992 한국미생물·생명공학회지 Vol.20 No.1

유기용매 이상계에서의 thermolysin을 사용하여 아스파탐 전구체 합성시 pH, 온도, 기질농도, 그리고 유기용매상에 대한 수용액상의 부피비(α) 등의 변화에 따른 기질의 분해 반응, 효소의 안정성, 그리고 Z-APM 합성에 미치는 복합적인 영향을 조사함으로써 반응조건의 최적화를 도모하였다. 유기용매 이상계에서의 L-PM·HCl의 자연분해는 수용액에서보다 휠씬 느리게 일어나며, 또한 α가 증가할수록 분해속도가 감소하는 것을 알 수 있었다. 효소의 안정성은 유기용매 이상계를 이용하였을 때 buffer에서 보다 월등 하게 좋았으며 특히 α=1인 경우 10일 후에도 90% 이상 높은 안정성을 나타내었다. α=1인 유기용매 이상계에서 Z-APM의 합성은 pH 5.5∼5.8에서 가장 수율이 높았는데, 이는 이상계에서 기질 및 산물의 분배계수로부터 에측했던 최적 pH 범위와 잘 일치하고 있음을 알 수 있다. 대체로 초기 기질농도가 높을수록 전환율이 증가하였으며 L-PM·HCl과 Z-Asp의 몰비가 2:1인 경우에 28시간 반응 후 90%정도의 수율을 나타냈다. 유기용매 부피비 변화에 따른 유기용매 이상계에서의 Z-APM 합성수율은 이론적으로 예측된 평형상수의 변화결과와 잘 부합함을 알 수 있었다. The synthesis of N-benzyloxycarbonyl-L-aspartyl-L-phenylalanine methyl ester(Z-APM), a precursor of aspartame, from N-benzyloxycarbonyl-L-aspartic acid(Z-Asp) and L-pheny-lalanine methyl ester hydrochloride(L-PM·HCl) was investigated in ethylacetate-MES buffer two-phase system using thermolysin. In organic two-phase system, the degree of spontaneous hydrolysis of L-PM·HCl was significantly reduced with increasing the volume ratio of organic to aqueous phase. Stability of thermolysin in organic two-phase system was found to be higher than that in MES buffer solution. More than 90% of initial enzyme activity was maintained after 10 days of incubation in case that the volume of organic phase was equal to that of buffer phase, while the half life of thermolysin was about 2 days in aqueous buffer solution. The results of partitioning of substrates and product in organic two-phase system showed that the difference in partition coefficients between substrates and product was maximum at pH 5.5. The optimal pH for Z-APM synthesis in organic two-phase system was found to be 5.5∼5.8, which is consistent with the value expected from the partition experiments. As the concentration of substrates was increased the conversion yield of Z-APM was increased with concomitant reduction of L-PM·HCl hydrolysis. In case that the concentration of L-PM·HCl and Z-Asp were 160mM and 80mM respectively, the conversion yield of Z-APM reached 90% after 28 hrs of reaction. The yield obtained at different volume ratio of organic phase compares well with the predicted equilibrium constant in biphasic system.