Flask culture of White Grifola frondosa for the optimization condition of production polysaccharides and its anti-inflammatory effect by high pressure extraction
Date Issued
2015
Date
2015
Author(s)
Chen, Ying-Wei
Abstract
The edible mushroom White Grifola frondosa is a Basidiomycete fungus belongs to the family Polyporaceae. For recent years, most studies related to Grifola frondosa are focusing on pharmacological and biological activities of polysaccharide. Polysaccharide extracted from the fruiting body and mycelium have been reported to have medical effects such as antitumor, hepato-protective activities and also can strengthen the immune system. In this study, White Grifola frondosa was cultivated in shaking flask under different media compositions and conditions in order to get the higher production of mycelial biomass and polysaccharide. Moreover, compared with the impact of mycelial polysaccharide by hot water and high hydrostatic pressure extraction and evaluated its anti-inflammatory activity in RAW 264.7 cell line. Among four kinds of carbon sources-Glucose, Fructose, Sucrose and Corn Starch, the result showed that Corn Starch was the most suitable carbon source for both mycelial biomass production. While Fructose can got more production of intracellular polysaccharide which was 20 grams per liter of medium. The use of Yeast Extract was the optimal nitrogen source than Malt Extract for both mycelial biomass and polysaccharide production. In addition, among four tested initial pH in cultural media, pH 6.0 was the optimal condition for the production of mycelial biomass and polysaccharide, followed by pH5. In terms of extraction, the crude extract of vacuum freeze drying mycelial polysaccharide obtained the higher extraction yield by hot water extraction than high hydrostatic pressure extraction, followed by 400 MPa but no significant difference between 50-200 MPa. Besides, the extraction of hot air drying mycelial by 500 MPa improved 20% than vacuum freeze drying by 400 MPa. Although there is no obvious enhancement on the crude extract of mycelial polysaccharide by High Hydrostatics Pressure extraction, it can improve the extraction. On the other hand, the crude extracts of mycelial polysaccharide (CEMP) by hot water and high hydrostatic pressure on the production of pro-inflammatory factors (NO, ROS) in the three lipopolysaccharide-stimulated inflammation models (preventive inflammation model, repairing inflammation model and co-treatment model) with RAW 264.7 cell line were evaluated. Effect of CEMP on the production of NO were measured by three models (preventive inflammation model, repairing inflammation model and co-treatment inflammation model). Cell viability results showed that crude extract of mycelial polysaccharide had no significant cytotoxicity in RAW 264.7 cell line. All of CEMP significantly inhibited or decreased the production of NO with dose-dependent manner on preventive and repairing inflammation models. Especially, the preventive inflammation model had the best effect on inhibition of NO production. However, all of CEMO on co-treatment inflammation model had no effect on inhibition of NO production. In conclusion, all of the above results demonstrated that different nutritions and environmental factors influenced mycelial growth and polysaccharide production. The finding may provide a useful information in the future for production of White Grifola frondosa on a large scale. Moreover, all of CEMP by hot water and high hydrostatic pressure extraction had better anti-inflammatory activity in preventive inflammation models… Therefore, CEMP can be applied to the development of functional food to increase the industrial utilities and to promote the additional values of White Grifola frondosa.
Subjects
White Grifola frondosa
polysaccharide
high hydrostatic pressure extraction
RAW 264.7 cell line
anti-inflammation activity
Type
thesis
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