• 약학회지
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• 제53권1호
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• pp.34-40
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• 2009

The glycosylation of therapeutic glycoproteins can affect their efficacy, stability, solubility, and half-life. Analyzing the composition of monosaccharides, such as that of neutral and amino sugars, is the first step for elucidating the structure of glycan attached to glycoproteins. In the present study, neutral and amino sugars of lectin obtained from Maackia fauriei were analyzed using an enzyme-linked lectinsorbent assay (ELLA) and high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Peroxidase-labeled lectins such as concanavalin A, Ricinus communis agglutinin, and soybean agglutinin were used for ELLA, since they specifically bind to the monosaccharide residue most frequently encountered in a glycan. The hydrosylate of lectin was prepared by treatment with trifluoroacetic acid, which resulted in the lectin mainly possessing the N-glycan consisting of 98.1 pmol Fuc, 342.1 pmol GlcN, 51.9 pmol Gal, 678.9 pmol Man, and 330.7 pmol Xyl. The present results demonstrate that ELLA and HPAEC-PAD are very effective methods for rapidly estimating the types and relative amounts of monosaccharides in intact glycoproteins.

• Journal of Applied Biological Chemistry
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• 제42권4호
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• pp.166-168
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• 1999

Inulo-oligosaccharides (IOS, $F_n$, n=2-6) were purified from enzymatic hydrolysates of water-soluble extract of Jerusalem artichoke tubers. Quantification of inulo-oligosaccharides was done using high pH anion exchange chromatography with pulsed amperometric detector (HPAEC-PAD) at the concentration range of 10-100 mg/L, which was compared with that of fructo-oligosaccharides (FOS, $GF_n$, n=1-7). Peak areas per mg IOS were higher than FOS at the same degree of polymerization (DP). Specific peak areas of IOS increased proportionally as DP increased up to six, in contrast to FOS showing no linearity.

• 대한본초학회지
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• 제22권4호
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• pp.261-270
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• 2007

Objective : The 5-HMF was not index material suitable to do the quality control of Rehmanniae Radix Preparata. In this study, We estimated the changes of oligosaccharide contents in Rehmanniae Radix Preparata using high-performance anion-exchange chromatography with pulsed amperometric detection(HPAEC-PAD). Methods : The analysis of oligosaccharide was conducted by HPAEC-PAD with Carbopac PA1, $250{\times}4mm$, 5um, and Carbopac PA1 guard column. Column temperature was kept at $30^{\circ}C$. Elution was carried out at 1000 ${\mu}l/min$ with 70mM NaOH and the injection volume was $10{\mu}l$. Each component was detected by PAD. Results : Nine constituents were found from merchandising Rehmanniae Radix Preparata(MR), while seven constituents were found in various processed Rehmanniae Radix Preparata. Not all constituents were defined but stachyose and raffinose were found in all cases. And The most common constituents of Rehmanniae radix was stachyose. In the course of processing, most of stachyose and raffinose were decreased. Stachyose was decreased slowly in the course of processing with rice wine(RR), amomi and rice wine(AR), and crataegi and rice wine(CR). However stachyose was decreased rapidly in the course of processing with fresh rehmannia juice(FR). The method with crataegi and rice wine(CR) showed the smallest decrease of stachyose. And processing method with crataegi and rice wine(CR) showed the most abundant amount for stachyose after the nineth processing. Conclusion : The changes of oligosaccharides in the course of processing were a very important direct barometers to do the quality control and set up a standard of Rehmanniae Radix Preparata.

• 한국미생물·생명공학회지
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• 제33권2호
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• pp.136-141
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• 2005

고형 양식사료에 포함된 질소와 인의 미생물학적 제거 공정을 알아보기 위하여 벤치규모의 실험을 수행하였다. CK-10과 CK-13 균주가 새우양식장의 물시료로부터 분리되었다. CK-10과 CK-13의 혼합배양에서 N/P의 동시제거 실험을 실시하였다. 그 결과, $400\;{\mu}M\;NH^{+}_4$ 와 $NO^{-}_2$는 12시간 이내에 제거되었고, $NO^{-}_3$는 36시간 이내에 각각 제거되었으며, $500\;{\mu}M\;PO^{3-}_4$ 36시간 이내에 제거되었다. CK-10과 CK-13 배양을 새우양식사료에서 용출된 N와 P의 제거에 적용하였다. HPAEC-PAD 시스템을 이용하여 양식사료의 당을 분석하였으며, glucose, galactose, galatosamine, mammonse, fucose 등의 여러 가지 당이 분석되었다. 세균에 의한 질소와 인 제거를 수행하기 위하여 인공 해수에서 $0.2\%$(w/v)의 양식사료를 용출시켰으며, 72 시간동안 용출된 질소의 양은 대략 $33.3\;{\mu}M\;NH^{+}_4,\;12.9\;{\mu}M\;NO^{-}_2.\;81.5\;{\mu}M\;NO^{-}_3,\;248\;{\mu}M\;PO^{-3}_4$였다. 혼합배양은 $0.2\%$ 사료에 포함된 질소와 인을 84시간 이내에 완전히 제거하였으나, 단일배양은 주어진 배양기간동안 질소와 인을 완전제거 하지 못하였다. 이 연구에서 CK-10과 CK-13 배양은 새우양식사료에서 유래하는 질소와 인을 효과적으로 제거하는 것이 입증되었다.

• Bulletin of the Korean Chemical Society
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• 제34권9호
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• pp.2663-2670
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• 2013

The purpose of stability testing is to provide evidence about how the quality of a drug varies with time under the influence of a variety of environmental factors. In this study, erythropoietin (EPO) was analyzed under different pH (pH 3 and pH 9) and temperature ($25^{\circ}C$ and $40^{\circ}C$) conditions according to current Good Manufacturing Practice (cGMP) and International Conference on Harmonisation (ICH) guidelines. The molecular weight difference between intact EPO and deglycosylated EPO was determined by SDS-PAGE, and aggregated forms of EPO under thermal stress and high-pH conditions were investigated by size exclusion chromatography. High pH and high temperature induced increases in dimer and high molecular weight aggregate forms of EPO. UPLC-ESI-TOF-MS was applied to analyze the changed modification sites on EPO. Further, normal-phase high-performance liquid chromatography was performed to identify proposed glycan structures and high pH anion exchange chromatography was carried out to investigate any change in carbohydrate composition. The results demonstrated that there were no changes in modification sites or the glycan structure under severe conditions; however, the number of dimers and aggregates increased at $40^{\circ}C$ and pH 9, respectively.

• 신재생에너지
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• 제3권1호
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• pp.38-45
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• 2007

To characterize thermo-chemical feature of sugar conversion of woody biomass, poplar wood ($Populus\;alba{\times}glandulosa$) powder was treated with supercritical water system. Supercritical water treatment (SCWT) was performed for 60 seconds at different temperatures (subcritical zone 350; supercritical zone $300,\;400,\;425^{\circ}C$) under two pressures $230{\pm}10atm$ as well as $330{\pm}10atm$, respectively, using flow type system. After separation of solid residues from SCWT products, the monomeric sugars in aqueous part converted from poplar wood powder were quantitatively determined by high performance anionic exchange chromatography [HPAEC] equipped with PAD detector and Carbo Pac PA10 column. As the temperature treated increased, the degradation of poplar wood powder was enhanced and ca 83% of woody biomass was dissolved into the water at $425^{\circ}C$. However, the pressure didn't help the degradation of biomass components. At subcritical temperature range, xylose was first formed by degradation of xylan, which is main hemicellulose component in hardwood species, while cellulose degradation started at the transition zone between sub and supercritical conditions and was remarkably accelerated at the supercritical temperature. In the supercritical water system the maximum yield of monomeric sugars amounts to ca. 7.3% based on oven dried wood weight at $425^{\circ}C$.

• ALGAE
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• 제22권3호
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• pp.247-252
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• 2007

Crude fucoidan was extracted from the sporophyll of Korean Undaria pinnatifida collected at a coastal area ofWando, Korea, mainly by dilute acid extraction, ethanol precipitation, CaCU Precipitation, with an yield of approxi-mately 3.9% in mass. It was further purified by DEAE-cellulose column chromatography and its chemical composi-don and in vitro anticoagulant activity was determined. The average molecular mass of the purified fucoidan wasestimated about 2.1 x 103 kDa by size-fractionation HPLC and it consisted of neutral sugar (52.34% in mass), uronicacid (26.2%), and sulfate esters (7.4%). From the HPAEC-PAD analysis, the monosaccharide composition of thepurified fucoidan was shown to be fucose, galactose, xylose, and mannose, with a molar ratio of 1, 0.2, 0.02, 0.15,respectively, demonstrating that major monosacd-iande was fucose (72.3% in mol percentage) and other sugars,xylose (1.5%), galactose (14.6%), and mannose (10.9%) were present as minor component. The results suggested thatthis fucoidan is a sulfated, U-type fucoidan. The activated partial thrombloplastin time (APTT) assay of the purifiedfucoidan showed that the purified fucoidan elicited anticoagulant activity in a dose-dependent manner. Five jUg ofsporophyll fucoidan delayed the blood clotting time up to 5 times than untreated control and also up to 1.5 timesthan the same amount of the commercial fucoidan, respectively. Although it is preliminary, these results suggestthat the fucoidan of Korean Undaria vinnatifida sporophyll would be promising candidates for the development ofan anticoaeulant.

• 한국식품영양과학회지
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• 제42권4호
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• pp.556-562
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• 2013

본 연구에서는 기존의 유기용매를 이용한 추출을 지양하고 환경 친화적이고 간단한 추출방법을 고안하여 톳의 다당류 성분(WSP-A)을 추출해내었고, WSP-A가 지질의 흡수에 미치는 효과를 알아보기 위하여 in vitro 상으로 lipase inhibition assay와 in vivo로 고지방 식이로 비만을 유도한 rat을 이용한 동물실험을 수행하였다. 그 결과 톳 유래 다당류는 alginate와 유사한 성분이지만 성분당 분석을 통하여 fucose가 주 당성분인 polysaccharide로 밝혀졌고, 이는 alginate보다 뛰어나고 안정한 lipase inhibitor로써 작용하였다. 또한 동물실험에서는 WSP-A를 포함하는 음용수로 제공한 것과 제공하지 않은 대조군은 무게 변화량에 유의적인 차이를 보일 뿐 아니라 비만 예방군과 비만 치료군 사이에서도 유의적인 차이를 나타내었다. 식이섬유를 이용한 연구와 비교해보았을 때, 톳 유래 다당류는 식이섬유 이상의 효과뿐만 아니라 뛰어난 lipase inhibitor로 체중감량을 나타내었고 이는 인간췌장유래 리파아제(human pancreatic lipase)의 저해활성과 더불어 인체 내에서의 작용 가능성을 짐작하고 있다. 따라서 톳 유래 다당류 성분은 식이성 섬유의 공급원 뿐만 아니라 추후에 효소활성 저해 효과에 대한 보다 명확한 작용기전을 밝혀낸다면 단순한 건강 기능성 식품 소재보다 lipase inhibitor로써 항비만 약물로 고부가가치를 창출해낼 수 있을 것으로 사료된다.

• Ocean and Polar Research
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• 제38권2호
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• pp.129-137
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• 2016

This study aimed to isolate and characterize sulfated polysaccharides (SPs) from Iridaea cordata and evaluate their anticancer activity. SPs of the Antarctic red seaweed were obtained by $CaCl_2$ (SP1) and ethanol precipitations (SP2) following diluted acid extraction at room temperature. Yields of SP1 and SP2 were approximately 14% and 23%, respectively, of the dry weight of red seaweed. The average molecular mass of the SP1 and SP2 was estimated about $1.84{\times}10^3$ and $1.42{\times}10^3kDa$, respectively, by size-fractionation High-Performance Liquid Chromatography (HPLC). From the High-Performance Anion-Exchange Chromatography-Pulsed Amperometric Detection (HPAEC-PAD) analysis, the main monosaccharide was galactose with glucose and fucose as minor components. The sulfate content of SP2 (40.4%) was slightly higher than that of SP1 (33.8%). The FT-IR spectra also showed characteristic band of carrageenan-like sulfated polysaccharides. Taken together the SPs are thought to be carrageenan-like sulfated galactan. The polysaccharides (SPs) from I. cordata exhibited weak antitumor activity against PC-3 (prostate cancer), HeLa (cervical cancer), and HT-29 (human colon adenocarcinoma). To our knowledge, this is the first data on biological activity of the Antarctic red seaweed I. cordata.

• Journal of the Korean Wood Science and Technology
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• 제42권6호
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• pp.740-746
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• 2014

꾸지뽕나무 미숙과를 50% 발효주정으로 추출하고, 농축한 추출용액을 n-hexane, chloroform, ethyl acetate, butanol로 순차적으로 분액하여 수용성 조추출물을 얻었다. 수용성 조추출물을 분취용 겔 투과 크로마토그래피(Gel Permeation Chromatography, GPC)를 실시하여 다당류(F1)를 분리하였다. F1을 가수분해하여 단당 성분 분석을 실시한 결과 glucose, galactose, arabinose, xylose 등 4가지로 구성되어 있었으며, 그 조성비는 6.1 : 1.8 : 3.1 : 1.0이었다. F1에 acetyl 기의 치환여부와 uronic acid의 존재 유무를 확인하기 위해 FT-IR로 측정하고 염산 가수분해 한 다음 HPLC 분석을 실시한 결과, 두 가지 모두 없는 것으로 분석되었다. 결합 위치 확인을 위해 permethylation, 산 가수분해, 환원, silylation을 순차적으로 실시하여 얻은 partially methylated alditol silylate (PMAS)들을 GC-MS 분석을 실시한 결과, 1,5-di-O-trimethylsilyl-2,3,4-tri-O-methyl xylose, 1,5-di-O-trimethylsily-2,3,4-tri-O-methyl arabinose, 1,4,5-tri-O-trimethylsily-2,3-di-O-methy arabinose, 1,3,5-tri-O-trimethylsily-2,4,6-tri-O-methyl glucose, 1,4,5-tri-O-trimethylsily-2,3,6-tri-O-methyl galactose, 1,4,5-tri-O-trimethylsily-2,3,6-tri-O-methyl glucose, 1,3,5,6-tetra-O-trimethylsily-2,4-di-O-methyl galactose, 1,3,5,6-tetra-O-trimethylsily-2,4-di-O-methyl glucose로 구성되어 있었고 그 조성비는 1.1 : 1.0 : 4.9 : 7.5 : 3.0 : 3.1 : 1.4 : 1.5였다.