• Title/Summary/Keyword: Solvent-free

Search Result 614, Processing Time 0.024 seconds

Antioxidative and Antimicrobial Activities of Aruncus dioicus var. kamtschaticus Hara Extracts (눈개승마(Aruncus dioicus var. kamtschaticus Hara) 용매 추출물의 항산화 및 항균활성)

  • Kim, Mi-Seon;Kim, Kyoung-Hee;Jo, Ji-Eun;Choi, Jong-Jin;Kim, Young-Jin;Kim, Jong-Hwan;Jang, Soon-Ae;Yook, Hong-Sun
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.40 no.1
    • /
    • pp.47-55
    • /
    • 2011
  • The solvent extracts of Aruncus dioicus var. kamtschaticus Hara, which were extracted by using several solvents with different polarities, were performed to investigate the antioxidant activities, whitening effect and antimicrobial activity. The content of total polyphenol of fractions from Aruncus dioicus var. kamtschaticus Hara extract showed the highest value ($335.88{\pm}2.26$ mg/g GAE) on ethyl acetate fraction. The ethyl acetate and n-butanol fractions were 0.06 mg/mL and 0.25 mg/mL as $IC_{50}$ values on DPPH radical scavenging, and $99.16{\pm}0.09%$ and $89.29{\pm}0.64%$ on ABTS radical scavenging activity, respectively. Also, reducing power and FRAP value were significantly higher on ethyl acetate fraction. The SOD like activity showed $80.76{\pm}0.61%$ on ethyl acetate and $72.34{\pm}0.79%$ on n-butanol. Tyrosinase inhibition activities (at 5 mg/mL) were $59.08{\pm}0.98%$ on ethyl acetate fraction. The chloroform fraction showed the strongest antimicrobial activities against B. cereus (14 mm), B. subtilis (12.5 mm), S. aureus (10.8 mm), E. coli (20.7 mm) at 0.1 mg/disc and the inhibition zone diameter of ethyl acetate fraction was 17.2 mm against E. coli at 0.5 mg/disc. The minimum inhibitory concentrations (MIC) of chloroform fraction against B. cereus and E. coli were 50 and $25{\mu}g$/mL, respectively. From these results, it is suggested that ethyl acetate and n-butanol fractions of Aruncus dioicus var. kamtschaticus Hara could be used as functional material for food additive ingredient and chloroform fraction could be suitable for the development of a food preservative.

Pharmacological Activity of Chaga Mushroom on Extraction Conditions and Immunostimulating Polysaccharide (추출조건에 따른 차가버섯 생리활성 및 면역활성 다당)

  • Baek, Gil-Hun;Jeong, Heon-Sang;Kim, Hoon;Yoon, Taek-Joon;Suh, Hyung-Joo;Yu, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.41 no.10
    • /
    • pp.1378-1387
    • /
    • 2012
  • To investigate the pharmacological activity of chaga mushroom (Inonotus obliquus) on extraction conditions, chaga was extracted using water (reflux at $50^{\circ}C$, decoction over $90^{\circ}C$, pressure at $121^{\circ}C$) or ethanol (reflux at 50, 70, or $90^{\circ}C$). When water extract was further fractionated into crude polysaccharide (IO-CP), yields of IO-CP (4.8~16.8%) were higher than those of ethanolic extracts (IO-E, 1.9~2.7%) at increased temperature. For antioxidant activity, crude polysaccharide (IO-CP-121) obtained by pressurized extraction showed the highest polyphenolic and flavonoid contents (35.10 mg TAE/g and 18.48 mg QE/g, respectively) as well as DPPH and ABTS free radical scavenging activities (26.08 and 27.99 mg AEAC/100 mg, respectively). Meanwhile, IO-CP-D (decoction) and IO-CP-50 (reflux) had more potent mitogenic effects (2.10- and 1.95-fold of saline control at 100 ${\mu}g/mL$) as well as intestinal immune system modulating activities (6.30- and 5.74-fold) compared to IO-CP-121, whereas ethanolic extracts showed no activity. Although no IO-CP showed cytotoxicity against RAW 264.7 cells at 0.1 mg/mL, IO-CP-121 significantly inhibited TNF-${\alpha}$ and NO production as pro-inflammatory factors in LPS-stimulated RAW 264.7 cells (29.2 and 63.5%, respectively). Ethanolic extracts also showed no cytotoxicity at 0.1 mg/mL, whereas inhibition of TNF-${\alpha}$ and NO production was significantly low compared to that of IO-CP-121. In addition, active IO-CP-D was further fractionated into an unadsorbed (IO-CP-I) and seven adsorbed fractions (IO-CP-II~VIII) by DEAE-Sepharose CL-6B column chromatography in order to isolate immunostimulating polysaccharide. IO-CP-II showed the most potent mitogenic effect and macrophage stimulating activity (4.51- and 1.64-fold, respectively). IO-CP-II mainly contained neutral sugars (61.86%) in addition to a small amount of uronic acid (2.96%), and component sugar analysis showed that IO-CP-II consisted mainly of Glc, Gal, and Man (molar ratio of 1.00:0.55:0.31). Therefore, extraction conditions affect the physiological activity of chaga, and immunostimulating polysaccharide fractionated from chaga by decoction is composed mainly of neutral sugars.

In Vitro Assay on Biological Characteristics of Different Extracts from Acorus calamus L. var angustatus (창포(Acorus calamus L. var angustatus) 추출물의 생리활성 검정)

  • Heo, Buk Gu;Park, Yong Seo;Yoo, Yong Kweon;Han, Tae Ho;Park, Yun Jum;Sin, Jang Sik;Cho, Ja Yong
    • FLOWER RESEARCH JOURNAL
    • /
    • v.16 no.3
    • /
    • pp.168-173
    • /
    • 2008
  • This study was conducted to determine biological characteristics of Acorus calamus L. angustatus by using aqueous, ethanol and methanol extracts from the leaves and the rhizomes of Acorus calamus L. angustatus plants. The highest total phenol contents were found in the extracts from the leaves of Acorus calamus L. angustatus ($68.4mg{\cdot}L^{-1}$) followed by rhizome ($49.3mg{\cdot}L^{-1}$). At $1,000mg{\cdot}L^{-1}$ the free radical scavenging activity of 1, 1-Diphenyl-2-picrylhydrazyl showed the highest activity in the extracts of the leaves and the rhizomes by 86.3% and 86.1%, respectively. Total flavonoid contents at $2,000mg{\cdot}L^{-1}$ extracts showed the greatest amount in the aqueous leaf extracts ($61.1mg{\cdot}L^{-1}$), and the ethanol rhizome extracts ($15.4mg{\cdot}L^{-1}$). Nitrite radical scavenging activity at $2,000mg{\cdot}L^{-1}$ was highest in the methanol extracts from leaves and rhizomes by 75.7% and 77.9%, respectively. Mushroom tyrosinase inhibition activity of leaves and rhizome from the Acorus calamus L. angustatus was very low, showing less than 1.3% regardless of the extract solvent, plant part, and concentration. These results indicated that methanol extracts from leaves exhibited higher biological activities than other extracts from rhizomes.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
    • /
    • 2016.05a
    • /
    • pp.19-20
    • /
    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

  • PDF