• Proceedings of the SCSK Conference
• /
• 2003.09a
• /
• pp.35-49
• /
• 2003

In this study, in an attempt to search for functional cosmetic ingredients from higher fungal, we have produced exopolysaccharides (GF-l, approximately carbohydrate 75％, protein 25％) and polysaccharide (GF-2) of mycelium extract, by submerged culture of Grifolafrondosa. For applications in anti-aging cosmetic field, we investigated the diverse biological activities. Antioxidant activity and inhibition of Matrixmetalloproteinases (MMPs) were investigated enzymatic assays by measuring the superoxide scavenging activity using xanthine-xanthine oxidase system and the proteolytic activity of MMPs using EnzChek Collagenase/Gelatinase kits, respectively. GF-l polysacchairde showed inhibition of superoxide radical by 90％ at a concentration of 0.2％ (w/v) and inhibition of collagenase by 45％ at 0.2％ (w/v). GF-2 polysaccharide of mycelium extract also exhibited good antioxidant activity. However, MMPs inhibition activity was relatively lower level compared to GF-l polysaccharides. The treatment of human dermal fibroblast (HDF) with GF-l and GF-2 polysaccharides increased the proliferation of fibroblast by approximately 23-25％ at a concentration of 0.5％ (w/v), also showed collagen synthesis increase in HDF by about 50％ at 0.5％ (w/v) compared to that of untreated control. We also report the moisturizing effects of polysaccharides in cosmetic products (O/W emulation) and its own ingredient, in vitro and in vivo. The GF-1 polysaccharide showed higher moisturizing ability than sodium hyaluronate, which is the most commonly used moisturizers ingredient. These results suggest the GF-l polysaccharide, protein-bound polysaccharide, may be used as an ingredient for new moisturizing and anti-aging cosmeceuticals.

• KSBB Journal
• /
• v.27 no.3
• /
• pp.177-185
• /
• 2012

Inonotus obliquus mushroom, which is a fungus belonging to Hymenochaetaceae family, is known to grow on birth trees in colder northern climates and to be a fungal parasite that draws nutrients out of living trees rather than from the ground. For the separation of protein-bound polysaccharide (PBP) from the culture broth and mycelium of Inonotus obliquus, three well known extraction methods namely hot water, ultrasound and microwave were used. The best extraction conditions to separate the PBP (64.94 mg/g) from mycelium by microwave were found to be for 1 hour and $150^{\circ}C$. The possibility for concentration of extracted PBP solution by using membrane was also studied. The extracted PBP solution was concentrated effectively by using an ultrafiltration membrane and the molecular weight cut off (MWCO) is 30 KDa. It was observed that a concentration by the ultrafiltration membrane is essential not only for the development of clean separation technology but also for enhanced production of PBP. As a result, we have shown that PBP in the final concentrated solution showed approximately 10 times higher than that in the crude solution by application of the developed separation systems. The separation yield of PBP was about 89.79% by gel filtration of purification steps and the purified product was confirmed to be PBP by using FT-IR.

• Korean Journal of Food Science and Technology
• /
• v.30 no.3
• /
• pp.702-708
• /
• 1998

The effects of mushrooms such as Lentinus edodes and Pleurotus ostreatus on anti-cancer activity through in vivo and in vitro experiments were powders of protein-bound polysaccharides in mushrooms were solubilized in 0, 5, 25 mg/kg saline, respectively and were used in vitro experiments. The in vivo experiments were carried out as followed: i) anti-cancer activities on Leukemia $(L_{1210})$, Hepatlicus cancer $(H_{22})$ and Sarcoma180/(S180), and ii) the effect on immune system through changes in intestine weight and the number of hemolytic plague forming cells. Protein-bound polysaccharides of all showed anti-cancer activity on $L_{1210}$ and fruit body of Lentinus edodes 25 mg/kg treatment group showed the highest inhibition rate (86%). Pleurotus ostreatus mycelial in medium of cultivate 25 mg/kg treatment. Fruid body of Lentinus edodes 25 mg/kg treatment group showed the highest inhibition rate (86% and 71%, respectively) on $H_{22}$ among them. The inhibition rates of fruit body and mycelial of Lentinus edodes 25 mg/kg treatment groups on S180 were 33.9% and 30.9%, respectively. Each samples of 50, 100, 200, $400\;{\mu}g/{\mu}L$ on in vitro cell toxicity test did not show significantly different cell death rates at P<0.05. In immune test, weights of liver and spleen were increased according to increase in conc. but were not significantly different at P<0.05. The weights of thymus were heavy in fruit body and mycelial of Lentinus edodes treatment group but were not significantly different at P<0.05. Hemolytic plague forming cells with antibody formation capability were significantly high in fruit body and mycelial of Lentinus edodes treatment samples.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.37 no.12
• /
• pp.1647-1653
• /
• 2008

This study used response surface methodology (RSM) in an effort to optimize the water extraction conditions of Hypsizigus marmoreus in order to increase cytotoxicity activity of the extract. A central composite design was applied to investigate the effects of independent variables, which included the extraction temperature ($X_1$), extraction time ($X_2$), the ratio of solvent to sample ($X_3$) on dependent variables of the extracts, including extraction yield ($Y_1$) and protein content ($Y_2$). The estimated optimal conditions were as follows: $51.3^{\circ}C$ extraction temperature, 8.2 hrs extraction time, and 46.7 mL/g of solvent per sample. The extract (CE) was extracted at optimal condition and crude polysaccharides (CPS) were obtained from CE by ethanol precipitation, dialysis, and freeze drying. Neutral (NPS) and acidic (APS) fraction of polysaccharides were seperated from CPS by ion chromatography. The growth inhibitory effects of the APS (0.5 mg/mL) on AGS human cancer cells were 73.97%. CPS showed the highest growth inhibitory effects on HepG2 human cancer cell at 0.5 mg/mL. However all fraction polysaccharides from Hypsizigus marmoreus showed lower than 20% growth inhibition on SW480 human cancer cell.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.4
• /
• pp.450-456
• /
• 1999

A high molecular-weight water-soluble fraction(PO) obtained by the ethanol precipitation of 0.1 N NaOH extracts of the mushroom Pleurotus ostreatus showed 82% anti-complementary activity for complement consumption hemolysis. The PO consisted of 42% carbohydrate (w/w), 50% protein (w/w), and 3% uronic acid (w/w). Fifty-eight percent of the anti-complementary activity decreased by periodate oxidation and 22% by protease digestion, suggesting that the sugar and protein moieties are essential for this activity. Two polysaccharide fractions, PO-IIIa-1 and PO-IIIa-2, with anti-complementary activity were isolated from the PO using DEAE-Sepharose FF followed by Sephadex G-75 and Sepharose CL-6B gel permeation chromatographies. The PO-IIIa-2 was found by HPLC to be nearly homogeneous, with the molecular mass of 531 kDa, and showed 96% $ITCH_{50}$ (inhibition against the total complement hemolysis of deionized water as the control) at a concentration of 1 mg/ml. This fraction contained galactose, mannose, fucose, and glucose with molar ratios of 1.75:1:0.65 and 0.59, respectively. The majority of galactose and mannose units in the PO-IIIa-2 were composed of TGalp1 ->, ->6Galp1->, ->2,6Galp1->, and ->Manp1->. The PO-IIIa-1 (molecular mass of 2000 kDa), exhibiting higher activity than the PO-IIIa-2, was further purified into two fractions, unbound proteoglycan (PO-IIIa-1A) and bound glucomannan (PO-IIIa-lB), by affinity chromatography using ConA-Sepharose CL-4B. The anti-complementary activity of each affinity purified fraction decreased as compared to that of the native PO-IIIa-1 fraction, indicating that the formation of complex between both polysaccharide fractions was necessary for full anti-complementary activity.

• Food Science and Preservation
• /
• v.11 no.1
• /
• pp.47-52
• /
• 2004

This study was conducted to investigate the characteristics of ultrafiltration and spray drying process for crude protein bound polysaccharide(CPBP) isolated from Agaricus blasei Murill. In ultrafiltration process, the permeate flux increased with the increase of operating pressure and temperature. The permeate flux declined continuously while the fouling materials were accumulated on the membrane as the operation time increased. In comparing of raw CPBP and filtered CPBP, the viscosity of CPBP treated UF was decreased and $\Delta$E value of treated samples was increased. Thermal efficiencies of spray drying process were increased by increasing inlet temperature, feed rate and feed concentration.

• Herbal Formula Science
• /
• v.8 no.1
• /
• pp.281-289
• /
• 2000

To examine the pharmacological activity of the polysaccharide extracted from phellinus linteus, we obtained the polysaccharide treated by the alkali and hot water, molecular weights of hot water was 10kD and 225 kD. Anticomplementary activity were highly observed in hot water fraction than alkali fraction. The tumor inhibition ratio of the polysaccharide extracted by hot water and alkali of Phellinus linteus against sarcoma 180 were 72.5% and 67.6%, respectively. These results suggest that protein bound polysaccharides extracted by a hot water and alkali of Phellinus linteus were similar to the pharmacological activities and chemical properties.

• KSBB Journal
• /
• v.10 no.5
• /
• pp.589-597
• /
• 1995

Protein-bound polysaccahrides(PBP) were isolated, purified, and characterized from the sawdust media after harvesting the fruit body of Flammulina velutipes. The yield of the crude PBP(Fr.CA) extracted from the sawdust media, was 0.367% relative to the original sawdust media. The total sugar and protein contents of Fr.CA were 19.8% and 23.8% respectively. Using the membrane filtration, the fraction of which the molecular weight is over 300 kDa(Fr.A) was isolated from the Fr.CA and the yield was 44.6% relative to the Fr.CA. This result indicates that high molecular PBP is the dominant components of the Fr.CA. The Fr.A was separated into three fractions (Fr.A-1, Fr.A-2 and Fr.A-3) whose yields are 5.8%, 8.5% and 13.2% respectively. These fractions were further purified using gel filtration, obtaining a single peak in each fraction that considered as pure PBP Among them, the yield of Fr.A-1-${\alpha}$ was 20.9% relative to the Fr.A-1, and the molecular weight was 800 kDa. Monosaccharide components such as glucose, galactose, mannose, xylose and fucose could be detected all fractions by a HPLC analysis. Especially in the Fr.A-1-${\alpha}$ fraction, the content of glucose and galactose appeared to be high.

• Journal of Mushroom
• /
• v.6 no.1
• /
• pp.27-31
• /
• 2008

The optimization of submerged culture conditions for mycelial growth and exopolysaccharide (EPS) production in an edible mushroom Tremella fuciformis were studied in shake flasks and bioreactors. The temperature of $28^{\circ}C$ and pH 8 in the beginning of fermentation in agitated flasks was the most efficient condition to obtain maximum mycelial biomass and EPS. The optimal medium constituents were as follows (g l-1): glucose 20, tryptone 2, $KH_2PO_4$ 0.46, $K_2HPO_4$ 1 and $MgSO_4H_2O$ 0.5. The fungus was cultivated under various agitation and aeration conditions in a 5L stirred-tank bioreactor. The maximum cell mass and EPS production were obtained at a relatively high agitation speed of 200 rpm and at an aeration rate of 2 vvm. The flow behavior of the fermentation broth was Newtonian and the maximum apparent viscosity (35 cP) was observed at a highly aerated condition (2 vvm). The EPS productivity in an airlift reactor was higher than that in the stirred-tank reactor. The EPS was protein-bound polysaccharides consisted of mainly mannose, xylose, and fructose. The molecular weights of EPS were determined to be $1.3{\sim}1.5{\times}10^6$.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.8
• /
• pp.1240-1247
• /
• 2006

The present study investigated the influence of the aeration rate and agitation intensity on the production of the mycelial biomass and antioxidative exopolysaccharide (EPS) in Ganoderma resinaceum. In submerged cultures with varying agitation speeds and aeration rates in a stirred-tank reactor, the maximum mycelial biomass and maximum EPS concentration were achieved at 50 rpm and 300 rpm, respectively. Under varying aeration rates, the highest amount of mycelial biomass (18.1 g/l) was accumulated at the lowest aeration rate (0.5 vvm) and the maximum EPS production (3.0 g/l) obtained at 1.0 vvm. A compositional analysis revealed that the five different EPSs were protein-bound heteropolysaccharides, consisting of 87.17-89.22% carbohydrates and 10.78-12.83% proteins. The culture conditions had a striking affect on the carbohydrate composition of the EPS, resulting in different antioxidative activities. All the EPSs showed strong scavenging activities against superoxide and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals, whereas no clear trend in antioxidative activity was observed against hydroxyl radicals and lipid peroxides. Although the precise reason for this difference is still unclear, the high glucose moiety of EPS is probably linked to its broad spectrum of antioxidative activity.