천년초에서 분리한 점질다당의 면역자극 활성

서이슬(Yi Seul Seo),신광순(Kwang-Soon Shin) 한국식품영양과학회 2012 한국식품영양과학회지 Vol.41 No.1

천년초(Opuntia humifusa)에 존재하는 점질다당의 새로운 이용방안을 모색하기 위해 천년초로부터 다당을 분리하여 면역자극활성을 측정하였다. 천년초의 주요 다당 CNC-E는 상업용 효소처리, 물 추출 및 ethanol 침전을 거쳐 조제하고 이들의 특성을 검토하였다. CNC-E는 분자량 약 700 kDa의 다당체로, 구성당 조성을 확인한 결과, arabinose, galactose 및 xylose를 높은 비율로 함유하고 있었으며 rhamnose와 fucose를 미량 함유하고 있었다. 한편 CNC-E는 비장세포 유래 림프구에 대해서는 비교적 높은 증식활성을 농도 의존적으로 나타냈다. CNC-E로 자극한 macrophage는 IL-6, IL-12, IL-10 및 TNF-α와 같은 cytokine의 생산을 농도 의존적으로 증진시켰으며 CNC-E를 mouse에 정맥 주사한 후 얻어진 NK세포는 Yac-1 종양 세포주에 대한 높은 치사작용을 보였다. 특히 CNC-E를 mouse 당 100μg 처리한 후 얻어진 NK세포는 미처리 대조군에 비해 3배 높은 치사활성을 나타냈다. CNC-E는 인체 초기 면역반응에 있어 중요한 역할을 담당하는 보체계에 대하여 농도 의존적인 활성을 나타냈으며, anti-human C3를 이용한 면역전 기영동실험과 Ca2+이온이 제거된 상태에서의 항보체 활성실험을 통해 천년초 다당 CNC-E는 고전경로와 부경로를 모두 경유하여 활성을 나타내고 있음을 확인하였다. 이상의 결과로부터 한국산 선인장 천년초에는 인체에 유익한 면역 활성 다당체를 다량 함유함을 알 수 있었다. To examine the new practical utilization of mucilages in Opuntia humifusa, the Korean pear cactus, the polysaccharides were isolated from O. humifusa, and immuno-stimulating activities were assayed. The main polysaccharide, CNC-E, was prepared by a commercial enzyme treatment, water extraction, and ethanol precipitation. The molecular mass of CNC-E was estimated to be about 700 kDa, and it consisted mainly of arabinose, galactose and xylose in addition to two minor sugars such as rhamnose and fucose. On the other hand, CNC-E showed considerably high splenocyte proliferation activity in a dose-dependent manner. Peritoneal macrophages stimulated with CNC-E produced cytokines such as IL-6, IL-12, IL-10, and TNF-α. The intravenous administration of CNC-E significantly augmented the cytotoxicity of natural killer (NK) cells against Yac-1 tumor cells. Especially, NK cells obtained from the mice treated with 100 μg of CNC-E showed threefold higher cytolytic activity than those of untreated mice. CNC-E also showed potent anti-complementary activity in a dose-dependent fashion. Identification of C3 activation products by the crossed immunoelectrophoresis using anti-human C3 and the anti-complementary activity of CNC-E in a Ca2+-free condition suggested complement activations by CNC-E that occur via both alternative and classical pathways. These results indicate that Korean pear cactus contains selected polysaccharides that provide immuno-stimulating activities beneficial to human health.

Isolation and Characterization of Immuno-stimulating Polysaccharide in Korean Black Raspberry Wine

( Yong Chul Hwang ),( Kwng Soon Shin ) 한국응용생명화학회 2011 Applied Biological Chemistry (Appl Biol Chem) Vol.54 No.4

To characterize polysaccharides existing as soluble form in Korean fruit wines, polysaccharides were isolated from Korean black raspberry wine, and their immuno-stimulating activities were examined. A complex pectic polysaccharide, RB-2 was purified from the crude polysaccharide (RB-0) in raspberry wine by subsequent size-exclusion chromatography. RB-2 had relatively potent macrophage activity and splenocyte mitogenic activity in vitro. The intravenous administration of RB-2 significantly augmented the cytotoxicity of natural killer cells against Yac-1 tumor cells. RB-2 consisted of 15 monosaccharides including rarely observed sugars in general polysaccharides, such as 2-O-methyl-fucose, 2-O-methyl-xylose, apiose, aceric acid, 3-deoxy-Dmanno-2-octulosonic acid, and 3-deoxy-D-lyxo-2-heptulosaric acid. Methylation analysis indicated that RB-2 comprised at least 20 different glycosyl linkages such as 3,4-linked fucose, 2,3,4-linked rhamnose, 3`-linked apiose, and 2,3,3`-linked apiose, characteristic of rhamnogalacturonan II (RGII). RB-2 was eluted as two peaks having a high (9,000 Da) and a low molecular mass (5,000 Da) on high performance size exclusion chromatography, and the peak of high molecular mass was down shifted to the low molecular mass under the acidic pH conditions less than 2.0. Results indicated higher molecular weight polysaccharide in RB-2 was mainly presented as an RG-II dimer, and Korean black raspberry wine contains selected polysaccharides, which provide immunostimulating activities beneficial to human health.

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.

복분자주에서 분리한 Mannan 다당의 면역증진 활성

이호(Ho Lee) 한국식품과학회 2012 한국식품과학회지 Vol.44 No.3

한국 전통발효음료 중에 존재하는 특이 다당류의 화학적 특성 및 생물활성을 규명할 목적으로, 농가에서 직접 발효한 복분자주로부터 다당류를 분리하고 이들의 각종 면역증강활성에 대해 검토하였다. 복분자주에 80% ethanol 침전을 행하여 얻어진 조다당획분 RB-0를 이용, 연속적인 3회의 column chromatography를 행하여, 복분자주의 주요 다당인 RB-1b-I으로 정제할 수 있었다. RB-1b-I획분은 HPLC상에서 대칭을 유지하는 단일 peak로 검출되었으며, 분자량은 약 180 kDa으로 평가되었다. 정제다당인 RB-1b-I의 구성당 조성을 확인한 결과, mannose가 76.8%로 주를 이루고 있었으며, 그 외에 galactose(15.8%), arabinose(3.8%), glucose(2.6%) 및 rhamnose(1.2%)의 순으로 함유되어 있었다. 이러한 결과는 복분자 중에 존재하는 주 다당이 발효 효모의 세포벽에서 기원한 mannan임을 추정하게 하였다. 한편 복분자주에서 정제한 다당 RB-1b-I은 in vitro상에서 macrophage의 활성화를 높은 비율로 유도하였으며, 비장세포에 대해 높은 증식능을 보였다. 또한 RB-1b-I을 정맥 투여한 경우, NK cell을 활성화하여 YAC-1 종양세포에 대한 세포독성을 증가시킴이 관찰되었다. 한편 고분자 정제 다당인 RV-1b-I은 비특이적 면역계에 있어 중요 역할을 담당하고 있는 보체계에 대하여 농도 의존적인 활성화 경향을 보였다. 이들은 Ca++ 이온이 제거된 상태에서의 항보체 활성과 antihuman C3를 이용한 2차원 면역전기영동에 의하여 C3 산물을 동정한 결과로부터 보체계의 classical pathway와 alternative pathway양 경로를 모두 경유하여 활성을 나타냄을 확인할 수 있었다. 이상의 결과로부터 복분자주에는 인체 건강에 유익한 면역증진 활성을 제공하는 다당류가 함유되어 있음을 알 수 있었다. Polysaccharides isolated from Korean black raspberry wine were examined for their chemical properties and immuno-modulating activities. The molecular mass of RB-1b-I, the main polysaccharide in black raspberry wine, was estimated as 180 kDa and it contained a significant proportion of mannose (76.8%) and 4 different minor component sugars such as galactose (15.8%), arabinose (3.8%), glucose (2.6%) and rhamnose (1.2%). This indicated that RB-1b-I was mainly present as a mannan, which had originated from the cell walls of fermenting yeasts. On the other hand, RB-1b-I induced high levels of macrophage activation as well as mitogenicity regarding murine splenocytes in vitro. The intravenous administration of RB-1b-I significantly augmented NK cytotoxicity against YAC-1 tumor cells. RB-1b-I also showed potent anti-complementary activity in a dose-dependent manner via both alternative and classical pathways. Results indicated that Korean black raspberry wine contains peculiar polysaccharides which provide beneficial immuno-stimulating activities for human health.

Isolation and Characterization of Immuno-stimulating Polysaccharide in Korean Black Raspberry Wine

Yong Chul Hwang,Kwang-Soon Shin 한국응용생명화학회 2011 Applied Biological Chemistry (Appl Biol Chem) Vol.54 No.4

To characterize polysaccharides existing as soluble form in Korean fruit wines, polysaccharides were isolated from Korean black raspberry wine, and their immuno-stimulating activities were examined. A complex pectic polysaccharide, RB-2 was purified from the crude polysaccharide (RB-0) in raspberry wine by subsequent size-exclusion chromatography. RB-2 had relatively potent macrophage activity and splenocyte mitogenic activity in vitro. The intravenous administration of RB-2 significantly augmented the cytotoxicity of natural killer cells against Yac-1 tumor cells. RB-2 consisted of 15 monosaccharides including rarely observed sugars in general polysaccharides, such as 2-O-methyl-fucose, 2-O-methyl-xylose, apiose, aceric acid, 3-deoxy-Dmanno-2-octulosonic acid, and 3-deoxy-D-lyxo-2-heptulosaric acid. Methylation analysis indicated that RB-2 comprised at least 20 different glycosyl linkages such as 3,4-linked fucose, 2,3,4-linked rhamnose, 3'-linked apiose, and 2,3,3'-linked apiose, characteristic of rhamnogalacturonan II (RGII). RB-2 was eluted as two peaks having a high (9,000 Da) and a low molecular mass (5,000 Da) on high performance size exclusion chromatography, and the peak of high molecular mass was down shifted to the low molecular mass under the acidic pH conditions less than 2.0. Results indicated higher molecular weight polysaccharide in RB-2 was mainly presented as an RG-II dimer, and Korean black raspberry wine contains selected polysaccharides, which provide immunostimulating activities beneficial to human health.

감식초에서 분리한 면역활성다당의 특성

황용철(Yong Chul Hwang),신광순(Kwang-Soon Shin) 한국식품과학회 2008 한국식품과학회지 Vol.40 No.2

한국 전통발효음료 중에 존재하는 특이다당류의 화학적 특성 및 생물활성을 규명할 목적으로, 농가에서 직접 발효한 감식초로 부터 다당류를 분리하고 이들의 각종 면역증강활성에 대해 검토하였다. 감식초에 80% ethanol 침전을 행하여 얻어진 조다당 획분 PV-0을 이용, 연속적인 3회의 column chromatography를 행하여, 감식초의 주요 다당인 PV-1b-I으로 정제할 수 있었다. PV-1b-I획분은 HPLC상에서 대칭을 유지하는 단일 peak로 검출되었으며, 분자량은 약 110 kDa으로 평가되었다. 정제다당인 PV-1b-I의 구성당 조성을 확인한 결과, mannose(46.8%), galactose(28.5%) 및 arabinose(19.1%)를 높은 비율로 함유하고 있었다. 또한 본 정제 다당은 β-glucosyl Yariv reagent와 강하게 반응하는 특성을 보여주었으며 이러한 사실로부터 PV-1b-I이 arabino-3,6-galactan 부위를 소유함을 확인할 수 있었다. 한편 감식초에서 정제한 다당 PV-1b-I은 in vitro상에서 macrophage의 활성화를 높은 비율로 유도하였으며, 비장세포에 대해 높은 증식능을 보였다. 또한 PV-1b-I을 정맥 투여한 경우, NK cell을 활성화하여 YAC-1 종양세포에 대한 세포독성을 증가시킴이 관찰되었다. 한편 고분자 정제 다당인 PV-1b-I는 비특이적 면역계에 있어 중요 역할을 담당하고 있는 보체계에 대하여 농도 의존적인 활성화 경향을 보였다. 이들은 Ca²? 이온이 제거된 상태에서의 항보체 활성과 anti-human C3 를 이용한 2차원 면역전기영동에 의하여 C3 산물을 동정한 결과로부터 보체계의 classical pathway와 alternative pathway 양 경로를 모두 경유하여 활성을 나타냄을 확인할 수 있었다. In this study, polysaccharides were isolated from Korean persimmon vinegar to characterize the polysaccharides existing as soluble forms within traditional Korean fermented beverages, and their immuno-stimulating activities were examined. Three successive chromatographies were used to purify the main polysaccharide in the persimmon vinegar, PV-1b-I, to homogeneity from the crude polysaccharide (PV-0). The molecular mass of PV-1b-I was estimated as 110 kDa and it contained significant proportions of mannose (46.8%), galactose (28.5%) and arabinose (19.1%). PV-1b-I strongly reacted with β-glucosyl Yariv reagent, suggesting the presence of an arabino-3,6-galactan moiety. PV-1b-I also induced high levels of macrophage activation and mitogenicity on murine splenocytes in vitro. The intravenous administration of PV-1b-I significantly augmented NK cytotoxicity against YAC-1 tumor cells. PV-1b-I also showed potent anticomplementary activity in a dose-dependent manner. Finally, C3 activation products were identified by crossed immunoelectrophoresis using anti-human C3 and the anti-complementary activity of PV-1b-I under Ca²?-free conditions, suggesting that this PV-1b-I causes complementary activations via both alternative and classical pathways. From these results, one can conclude that Korean persimmon vinegar contains select polysaccharides in addition to healthy components, and these polysaccharides appear to provide immuno-stimulating activities beneficial to human health.

Isolation and Characterization of Immuno-stimulating Polysaccharide in Korean Black Raspberry Wine

Hwang, Yong-Chul,Shin, Kwang-Soon The Korean Society for Applied Biological Chemistr 2011 Applied Biological Chemistry (Appl Biol Chem) Vol.54 No.4

To characterize polysaccharides existing as soluble form in Korean fruit wines, polysaccharides were isolated from Korean black raspberry wine, and their immuno-stimulating activities were examined. A complex pectic polysaccharide, RB-2 was purified from the crude polysaccharide (RB-0) in raspberry wine by subsequent size-exclusion chromatography. RB-2 had relatively potent macrophage activity and splenocyte mitogenic activity in vitro. The intravenous administration of RB-2 significantly augmented the cytotoxicity of natural killer cells against Yac-1 tumor cells. RB-2 consisted of 15 monosaccharides including rarely observed sugars in general polysaccharides, such as 2-O-methyl-fucose, 2-O-methyl-xylose, apiose, aceric acid, 3-deoxy-Dmanno-2-octulosonic acid, and 3-deoxy-D-lyxo-2-heptulosaric acid. Methylation analysis indicated that RB-2 comprised at least 20 different glycosyl linkages such as 3,4-linked fucose, 2,3,4-linked rhamnose, 3'-linked apiose, and 2,3,3'-linked apiose, characteristic of rhamnogalacturonan II (RG-II). RB-2 was eluted as two peaks having a high (9,000 Da) and a low molecular mass (5,000 Da) on high performance size exclusion chromatography, and the peak of high molecular mass was down shifted to the low molecular mass under the acidic pH conditions less than 2.0. Results indicated higher molecular weight polysaccharide in RB-2 was mainly presented as an RG-II dimer, and Korean black raspberry wine contains selected polysaccharides, which provide immunostimulating activities beneficial to human health.

유자 에탄올 추출물의 면역력 증진 효과

양지원(Jiwon Yang),전혜린(Hyelin Jeon),유양희(Yang Hee You),김진영(Jin Young Kim),최효경(Hyo-Kyoung Choi),최경철(Kyung-Chul Choi),전우진(Woo Jin Jun),윤호근(Ho-Geun Yoon) 한국식품영양과학회 2015 한국식품영양과학회지 Vol.44 No.9

본 연구에서는 유자가 선천성 면역력에 미치는 효과를 알아보기 위하여 유자 30% 주정추출물(CJE)을 사용하였으며 선천성 면역에 중요한 역할을 하는 대식세포를 이용해 실험하였다. CJE는 마우스 대식세포인 RAW264.7에서 1,000 μg/mL의 최고 농도까지 세포독성을 보이지 않았고, 전사인자 NF-κB와 염증성 매개물질인 COX-2, PGE2의 활성 및 발현 증강에 영향을 미치며, 특히 300 μg/mL의 농도에서부터 유의적 차이를 보이는 것으로 확인되었다. 산화질소 생성능과 대식세포에서 분비되는 사이토카인인 TNF-α, IL-1β의 발현을 대조군에 비해 농도 의존적으로 증가시킨다는 결과를 얻었으나, IL-6에서는 통계적으로 약간의 유의성이 있는 증가를 보였고 IL-10은 정상대조군에 비해 거의 유의적인 차이를 보이지 않았다. CJE는 또한 NK 세포의 활성을 농도의존적으로 증가시키고 비장세포의 증식능도 농도 의존적으로 증가시킨다는 것을 확인하였다. 이러한 결과로 미루어보아 CJE는 인체의 대식세포 활성의 증가를 통해 선천성 면역력을 증가시킬 것으로 판단된다. Citrus junos Sieb. ex Tanaka has been traditionally called Yuza in Korea and is used as a cuisine material or tea as well as medicinal herb. In this study, we evaluated the immune-enhancing effect of Citrus junos ethanol extract (CJE) on RAW264.7 mouse macrophage and primary immunocytes. CJE treatment showed increased macrophage activity in a dose-dependant manner. CJE also enhanced natural killer (NK) cell activity. We measured lactate dehydrogenase (LDH) level as a measurement of NK cell cytotoxicity against YAC-1 lymphoma cells. CJE treatment showed an increased LDH level in a dose-dependent manner. Finally, we evaluated the effect of CJE on mouse primary splenocyte proliferation. CJE treatment slightly increased splenocyte proliferation compared to the control. The results of this study suggest that CJE can help immune function via macrophage cytokine production, increased NK cell activity, and splenocyte proliferation.

유전자조작, 균주분리 청국장으로부터 면역증강활성이 우수한 Bacillus pumilus JB-1의 분리 및 분리균의 청국장 발효특성

권하영 ( Ha Young Kwon ),김영숙 ( Young Sook Kim ),권기석 ( Gi Seok Kwon ),권정숙 ( Chong Suk Kwon ),손호용 ( Ho Yong Sohn ) 한국미생물생명공학회 2004 한국미생물·생명공학회지 Vol.32 No.4

Characterization and Immunostimulating Activity of a Water-soluble Polysaccharide Isolated from Haematococcus lacustris

박제권,김지훈,이철균,Andriy Synytsya,조항수,김성욱,박주웅,박용일 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.6

A water-soluble polysaccharide was isolated and purified from the culture filtrate of the photosynthetic green microalgae Haematococcus lacustris by 75% ethanol precipitation and Sepharose CL-6B column chromatography. The molecular mass of the purified polysaccharide (named HCP) was estimated to be approximately 135 kDa by sizeexclusion HPLC and its monosaccharide composition was galactose, glucose and mannose at a relative molar ratio of 2.0, 1.0, and 4.1, respectively, suggesting that HCP is a galactomannan. Fourier-transform infrared and elemental analysis revealed that the purified HCP contains sulfate esters by 1.08% (in mass) and no detectable level of protein. The HCP significantly stimulated murine macrophage RAW264.7 cells to secrete the pro-inflammatory cytokine,TNF-α, in a dose-dependent manner and also enhanced the expression of COX-2 and iNOS genes at a concentration of lower than 10 μg/mL HCP. These results indicated that the sulfated HCP of H. lacustris has potent early innate immune stimulating activities.