| Summary: | 博士 === 國立中興大學 === 食品暨應用生物科技學系所 === 102 === The medical fungus Antrodia salmonea (or Taiwanofungus salmoneus), a newly identified species of the genus Taiwanofungus, which grows on the empty rotten trunk of Cunninghamia konishii in Taiwan. The fruiting body of A. salmonea has been used in the food remedy of diarrhea, abdominal pain, hypertension, itchy skin, and liver cancer and is also used as a detoxicant in Taiwanese folk medicine. To date, there were several newly compounds were isolated from the basidiomata of A. salmonea, whose studies displayed anti-oxidative effect and anti-inflammatory activities in activated inflammatory cells. With the rapidly increased standard of living, economic growth, and The improved medical sciences/techniques, people live longer than before. concurrent chronic and age-related diseases including cardiovascular disease, osteoporosis, arthritis, type II diabetes, certain cancers, and functional decline are increasing with the senior people and thereby, geriatrics is becoming a major issue to the public. beneficial to public health. Therefore, the aim of this study is to prepare the The research reported herein was to develop the Furthermore, the new products from medicinal mushrooms using grains as substrate, which are molecular identification method for A. salmonea strain. research was designed to evaluate the optimal conditions for A. salmonea to grow in grains were found including temperature, water supplement, initial pH value, inoculation rate, nitrogen and carbon sources; and fermented products including A. salmonea fermented oat (ASFO), wheat (ASFW), embryo rice (ASFE) and buckwheat (ASFB) were prepared. The proximate composition, taste components including total soluble sugars, free amino acid and 5′-nucleotides were determined. In addition, the research was focused on the antioxidant properties, and anti-inflammatory activity of mycelia and mycelium colonized grain products. Finally, the research was to study histopathology and biochemistry analysis in a 28-day toxicological assessment of A. salmonea fermented oat in BALB/c mice.
In order to know the biological origins of used strain, the research was to study molecular identification method for A. salmonea based on the nucleotide sequences of ITS1-5.8S-ITS2 rDNA. The results showed that total length of the ITS1-5.8S-ITS2 rDNA sequences from used strain was 808bp. In BLAST search, the first 4 sequences that have most identity of the published sequence in gene bank compared with the ITS1-5.8S-ITS2 rDNA of used strain were A. salmonea and they showed 99% maximum identity, except for one of A. camphorata strain. Furthermore, the total length of LSU rDNA D1-D3 In BLAST search, the first 7 sequences that have most sequences was 913bp. identity of the published sequence in gene bank compared with the LSU rDNA D1-D3 of used strain were A. salmonea and they showed 99% maximum identity. Based on the results and morphology of used strain, we could confirm the used strain was A. salmonea.
A solid state fermentation method is provided which effective for both small and large scale fungal cultivation. evaluate the mycelial biomass. The research reported herein was to study the optimal conditions for of A. salmonea grow in the selected grains, by With regard to the optimal conditions were 25°C, 28 days and 10% incubation rate with carbon and nitrogen sources being 1% fructose and 2% soybean meal for buckwheat, oat and wheat, except 1% yeast extract for embryo rice. mg/g.
Furthermore, the non-volatile components and physiological activity in the A. salmonea mycelia and A. salmonea colonized products were studied. A. salmonea mycelia were high in contents of crude fat, ash and protein and A. salmonea coloniezed in grains products were high in contents of carbohydrate.
The total sugar content of A. salmonea mycelium (51.99 mg/g) was higher than ASFB (34.24 mg/g), ASFW (29.4 mg/g), ASFR (28.88 mg/g) and ASFO (25.68 mg/g). respectively. Contents of total free amino acids were 13.13 and 6.97~12.99 Contents of total and flavor 5''-nucleotides from A. salmonea The equivalent umami concentrations [mg mg/g for A. salmonea mycelia and A. salmonea fermented products, mycelia (9.41 and 7.74mg/g) were higher than A.salmonea fermented products (5.82~7.56 and 4.0 ~5.46 mg/g). monosodium glutamate/100 g] were found in the descending order of: mycelia (167.05) > ASCO (80.83) > ASCE (63.99) > ASCW (13.63) > ASCB (5.62). With regard to physiologically active components, contents of ergosterol , ergothioneine and adenosine of A. salmonea mycelia were 3450, 320.9 and 529.8 µg/g, respectively. The fermented products contents of three phytochemicals GABA, lovastatin and ergothioneine were 90.93-170.23, 1.06-6.60 and 81.01-119.77 mg/kg, respectively. Overall, A. salmonea mycelia and fermented products possessed highly intense umami taste.
In addition, the antioxidant properties of hot water and 70% ethanolic extracts from A. salmonea mycelia and the four fermented products were investigated. The contents of potential antioxidant components in these The 70% ethanolic extracts were more extracts were also determined. effective than hot water extracts from A. salmonea mycelia in antioxidant activity and for mycelia of A. salmonea, most EC50 values were less than 6 mg/ml except for the hot water extract in antioxidant ability. For mycelia fermented products of A.salmonea, most EC50 values were less than 8 mg/ml. Overall, both extracts from A. salmonea mycelia and mycelia fermented products were good in antioxidant properties.
In the anti-inflammation activity test, we investigated the effect of 70% ethanolic extracts from A. salmonea fermented products ASFB, ASFO, ASFR and ASFW on LPS-induced NO and cytokines (TNF-α, IL-1β and IL-6) production in RAW 264.7 cells. At 500 μg/mL, the 70% ethanolic extracts With regard to the effect of At 500 from A. salmonea fermented products (41.02~46.44%) had more effective inhibition on NO production in RAW 264.7 cells. samples on LPS-induced TNF-α production in RAW 264.7 cells. μg/mL, the 70% ethanolic extracts from A. salmonea fermented products had (12.55~15.41%) a slight inhibition on TNF-α production in RAW 264.7 cells. In addition, the mycelia fermented products (32.85~40.3%) possessed a significant inhibition on IL-1β production in RAW 264.7 cells and their effective concentrations were 500 μg/mL. With regard to the effect of samples on LPS-induced IL-6 production in RAW 264.7 cells, 70% ethanolic extracts from the products had 32.85~40.3% of inhibition effect at concentration of 500 μg/mL. activity.
Overall, mycelia and filtrate of A. salmonea mycelia and fermented products contained abundant nutritional components, essential amino acids, umami components, physiologically active components, antioxidant properties and anti-inflammatory activity. Accordingly, A. salmonea might be potential antioxidants and anti-inflammatory agents for using in food products and could be developed as a new dietary supplement and functional food. A. salmonea mycelium fermented oats 1%, 2.5%, 5% and 1.66% A. salmonea mycelium mixed into the feed fed for 28 days, the each treatment groups relative organ weight and liver, heart, kidney, spleens H&E sections showed that due to feeding
A. salmonea mycelium fermented oats 1%, 2.5%, 5% and 1.66% A. salmonea mycelium were did not lead to tissue lesions. Also the concentrations of serum chemistry parameters in BALB/c mice fed with different substituted feed of A. salmonea mycelium fermented oat and 1.66% A. salmonea mycelium for 28 days were resulted have not toxicity.
In addition to the 7% A. salmonea mycelium, 7% four kinds of fermented products to replace part of the flour added to the dough, to make A. salmonea fermented buckwheat bread (ASFB bread), A. salmonea fermented oat bread (ASFO bread), A. salmonea fermented embryo rice bread (ASFR bread) and A. salmonea fermented wheat bread (ASFW bread) thus obtained were analyzed for its proximate composition, physicochemical properties and sensory evaluation as compared to the white bread control. are significantly higher those other breads. Crude fiber of A. salmonea bread (1.69%) and ASFO bread (1.90%) in proximate composition With regard to physicochemical properties, all breads specific volume were passing grades. In the analysis of free amino acids and nucleotides, A. salmonea mycelium bread, ASFB bread and ASFO breads taste components components were significantly more than white bread. Analysis of consumer preferences on sensory evaluation, although A. salmonea bread due to have a special color and flavor to reduce preference of consumers, but the A. salmonea fermented products breads were no significant difference in the taste and also have similar acceptance with white bread. In conclusion, addition of A. salmonea mycelium and A. salmonea fermented products to dough to make bread increased nutrition values and provide a new alternative of functional baked products to consumers.
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