| Antrodia Cinnamomea is famous in Taiwan as "niu-chang-chi" or "niu-chang-ku". Many physiological functions of A. cinnamomea cultures, including hepato-protective, anti-inflammatory, antioxidant, and anti-cancer functions were found. Among many physiological functions, triterpenoids was paid much more attention. Furthemore, the triterpenoids content has been as the important indicator of A. cinnamomea strain screening and production or quality control. In this study, the solid-state and submerd culture condition of A.cinnamomea was optimized to increase triterpenoids content, and the physiological functions of solid-state and submerd cultures were compared.The spectrophotometric quantitative method was optimized to measure the content of triterpenoid of Antrodia Cinnamomea by using vanillin-perchloric acid as the chromogenic agent. The reaction conditions were further optimized.1.0000 g fermented product was extracted with 50 mL 95% ethanol at 50℃in water bath for 1.5 h, the filtrate was used for analysis of total triterpenoids by colorimetry. Taked 0.10mL extraction, added vanillin solution 0.20 mL; added 0.50 mL perchloric acid to each tube, mixed and heated at 60℃in water bath for 20 min. After being cooled for 15 min, the sample was added 5.0 mL glacial acetic acid, finally measured absorbance for each sample at 550 nm. The method operated more conveniently, and the result was more repeatable and stabler.For the purpose of increasing the triterpenoids production, the submerged culture condition of A. cinnamomea was optimized by the approach of one variable at a time. The optimized medium was glucose 60 g/L, soybean hydrolyzate 40 mL/L, corn steep liquor 20 mL/L, MgSO4·7H2O 0.50 g/L, Na2HPO4 0.50 g/L. The optimized conditions were showed as following:inoculum 12.5%, temperature 28℃, rotation speed of 120 r/min, pH 5.5.Moreover, the factors that obviously affected the biomass and triterpenoids production of A. cinnamomea were optimized by Uniform Design and RSM. The optimal results:glucose 43.6 g/L, soybean hydrolyzate 75.7 mL/L, corn steep liquor 40 mL/L, MgSO4·7H2O 0.50 g/L, Na2HPO40.50 g/L. The optimized conditions were showed as following:inoculum 12.5%, temperature 28℃, rotation speed of 120 r/min, pH 4.75. The experimental results showed that the biomass and triterpenoid production under the optimal culture condition reached to 12.05 g/L and 45.04 mg/g. It was 16.5 and 2.16 times times than the pre-optimized yield of biomass 0.73 g/L and triterpenoid production 20.9 mg/g.For the purpose of increasing the triterpenoids production, the solid-state culture condition of A. cinnamomea was optimized by the approach of one variable at a time. The optimized conditions were millet 105g, NH4Cl 0.3 g, water 78 mL, inoculum size 45 mL and temperature 28℃.Moreover, the factors that obviously affected the biomass and triterpenoids production of A.cinnamomea were optimized by Uniform Design and RSM. The optimal results:millet 120g, NH4Cl 0.22 g, water 48 mL, inoculum size 55mL and temperature 28℃. The experimental result showed that the triterpenoid production under the optimal culture condition reached to 82.44 mg/g(dry weight). It was increased to 1.56 times than the pre-optimized yield, which was 5 times than the triterpenoid production in submerged culture in reported.Comparison of the major metabolites and content in solid-state cultivation and liquid-state cultivation by A. cinnamomea, the results showed that the total triterpenoids content in the products from SSC were 84.44 mg/g, which was one time higher than that from LSC. Furthermore, the triterpenoids kinds were obviously higher than that from LSC. The polysaccharides content from SSC was more than 5-fold higher than that from LSC. However, theβ-D-glucose content in submerged culture was 0.01 g/g, which was 2-foldβ-D-glucose content in solid-state culture. The content of fatty acids, amino acids and volatile substances, anti-oxidation capability and reducing power were higher than those in submerged culture.
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