Maca roots (Lepidium meyenii) are an important medicinal herb that have long been used by the Andes-indigenous peoples and South Americans. In Korea, recently, it has attracted attention as a health food material because of nutritional values and physiological activities. The purpose of this study was to investigate the industrial applicability of maca (red and golden varieties; R&G) as immunostimulating materials. In the macrophage stimulating assay using RAW 264.7 cells at 125~500 μg/mL of non-cytotoxicity doses, G-HW showed the most potent production of TNF-α, IL-6 and nitric oxide compared to red maca or the other extracts. The general component analysis results showed that all extracts comprised more than 90% neutral sugars with small amounts of uronic acid and protein. Meanwhile, component sugar analysis showed the difference in the content of uronic acids of cold- and hot-water extract. Additionally, the further fractionation of G-HW into crude polysaccharide (G-CP) greatly enhanced the macrophage stimulating activity, and G-CP contained macromolecules over 144 kDa, comprised mainly of glucose and galacturonic acid (51.0 and 34.9%). In conclusion, crude polysaccharide from maca showed industrial applicability as immunostimulating material, and especially golden maca showed higher macrophage stimulating activity than red maca.

After ethanol (BM-E and RW-E) and hot-water (BM-HW and RW-HW) extracts were fractionated from two herbal mixtures (BM and RW), their physiological activities were investigated. All extracts consisted of more than 50% of neutral sugar, with their total polyphenol levels higher than flavonoid levels. Radical scavenging activities of EtOH extracts remained significantly higher compared to that of hot-water extracts, and in particular, RW-E showed consistently higher antioxidant activity than BM-E. When anti-inflammatory activities of the extracts were evaluated by LPS-stimulated RAW 264.7 cells at 10~500 μg/mL non-cytotoxicity doses, BM-E showed significantly higher levels of TNF-α, IL-1β, IL-6, and nitric oxide inhibitory activity than those of hot-water extracts and RW-E. Murine peritoneal macrophage cells were shown to be enhanced in crude polysaccharides (BM-CP and RW-CP fractionated from BM-HW and RW-HW) compared to hot-water extracts and polysaccharide K (PSK, positive control). Especially, RW-CP exhibited higher activity than BM-CP, and component sugar analysis showed that BM-CP mainly contained galacturonic acid, glucose, arabinose, galactose, and xylose (34.5%, 33.9%, 16.1%, 7.1%, and 6.3%, respectively), whereas RW-CP showed different measurements (29.5%, 59.2%, 5.0%, 4.5%, and 0.2%). In conclusion, two herbal mixtures could contain varying sets of physiological activities dependent on different extraction and fractionation methods.

To obtain the immunomodulating polysaccharide from chaga mushroom (Inonotus obliquus sclerotia, IO), crude polysaccharide fractions (IO-M-CP and IO-CP, respectively) prepared from hot-water extract (IO-W) of I. obliquus by EtOH precipitation after MeOH reflux or not. After IO-W was re-dissolved in water followed by EtOH addition in the case without MeOH reflux, EtOH mixture was fractionated into EtOH-soluble (IO-E) and crude polysaccharide (IO-CP). In the meanwhile, MeOH-soluble fraction (IO-M) was separated from IO-W after MeOH reflux. The residue was dissolved in water and was added by EtOH, and then EtOH mixture was also fractionation into EtOH-soluble (IO-M-E) and crude polysaccharide (IO-M-CP). As a result of the macrophage stimulating activity of these fractions, IO-CP and IO-M-CP showed significantly increased cell proliferation and cytokines production than IO-W. Particularly, IO-M-CP promotes the production of IL-12 more than IO-CP. In the splenocytes proliferating activity and intestinal immune system modulating activity through Peyer’s patch, both of 2 crude polysaccharide fractions were significantly promoted in cell proliferation and cytokines production than IO-W, and IO-M-CP was more potent than IO-CP in IL-2 production from splenocytes and GM-CSF production (10 μg/mL) in Peyer’s patch cells. In addition, immunomodulating polysaccharide fractions (IO-M-CP and IO-CP) prepared from IO-W by EtOH precipitation with or without EtOH reflux showed no significant difference in the chemical composition and component sugar. These results suggested that MeOH reflux might exclude low-molecular weight materials from IO-W and consequently increase the immunomodulating activity of IO-M-CP. Therefore, it was confirmed that immunomodulation of polysaccharide prepared from hot-water extract of chaga mushroom was enhanced by fractionation including MeOH reflux and EtOH precipitation.

In this study, we examined immuno-modulatory activities of crude polysaccharides from wild ginseng adventitious roots (WGAR). The crude polysaccharide (WGAR-CP) was isolated from WGAR by hot water extraction, ethanol precipitation, and dialysis. The major constituents in WGAR-CP were neutral sugar (64.77%), and uronic acid (34.32%). WGAR-CP demonstrated anti-complementary activity dose-dependently. The immuno-modulatory effects of WGAR-CP were also analyzed by measuring nitric oxide and cytokines in the supernatants of mouse peritoneal macrophages. Mouse peritoneal macrophages stimulated with WGAR-CP produced nitric oxide and various cytokines such as interleukin (IL)-6 and IL-12 in a dosedependent manner. In conclusion, WGAR-CP may have immuno-modulatory activities by activating a complementary system and macrophages, which produces cytokines.

흑목이 버섯의 자실체로부터 열수추출에 의해 얻은 조다당 분획물은 흑갈색 색소를 함유하였으므로 활성탄에 의한 탈색공정이 검토된 바 있다. 하지만 활성탄에 흡착된 색소 성분의 회수 및 활성탄의 재사용이 필요하였으므로 용매추출법을 이용한 용출 실험을 실시하였다. 증류수, 중성용매(3종) 및 알칼리 용매(4종) 등 8종 용매에 의한 1단 침출 실험 결과, 색소성분의 용해성은 1 M KOH 용액이 가장 우수하였다. 이러한 최적 용매인 1M KOH 용액을 이용하여 용액의 부피 및 시간에 따른 용출효과를 조사한 결과, 용액의 부피는 활성탄 1 g당 45 mL에서 최적의 용출량을 보였으며, 10분 이내에 빠르게 용출평형에 도달하였고, 용출공정의 용출속도는 2차 속도식으로 나타낼 수 있었다. 또, 25oC에서 1 M KOH 용액에 의한 다단침출의 실험 결과, 7단계 침출에서 최종 용출효율은 88.9%이었다. 그러나 온도가 증가할수록 용출효율이 급격히 높아져 95oC에서는 단일용출만으로 82.6%의 용출효율을 보였다. 한편, 용출 후의 재사용된 활성탄의 흡착효율(75.3%)은 신선한 활성탄(78.5%)과 거의 비슷하였고, 다당 정제율도 거의 비슷하여(1.21-1.25배) 재생 가능함을 보였다. 따라서, 흑목이 버섯의 흑갈색 성분의 탈색 후 흡착된 활성탄으로부터 용매추출을 이용한 효율적인 색소성분의 회수 및 활성탄 재생 공정의 가능성이 확인되었다.