Enhanced Yield of Bioactive Compounds and Antioxidant Activities in Four Fermented Beans of Phellinus linteus Strains (Agaricomycetes) by Solid-State Fermentation.

Phellinus linteus is a famous medicinal mushroom which exhibits various biological activities. This study aimed to investigate the effects of solid-state fermentation by Ph. linteus on the yield of bioactive compounds and antioxidant activities of beans. Four bean substrates were prepared and inoculated with inoculum of three strains of Ph. linteus, respectively. During the cultivation, the harvested samples were dried, grounded, extracted, and determined the contents of bioactive compounds and antioxidant activities. The results indicated that the mung bean fermented by Ph. linteus 04 had the highest polysaccharide content (98.8 mg/g). The highest total phenolic and flavonoid contents were in fermented soybeans by Ph. linteus 03 (15.03 mg gallic acid equivalents/g and 63.24 mg rutin equivalents/g, respectively). The 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activities of hot water extracts were higher than those of ethanolic extracts for fermented beans by three Ph. linteus strains. However, the superoxide anion radical scavenging ability of ethanolic extracts was higher than those of hot water extracts in the fermented beans of the three strains. The ferrous ion (Fe2+)-chelating abilities of hot water extracts were higher than those of ethanolic extracts in fermented beans by Ph. linteus 03 and 04. In contrast, ethanolic extracts were higher than hot water extracts in fermented beans by Ph. linteus 06. Overall, these results indicate that the fermentation by Ph. linteus strains increased the bioactive compounds and antioxidant activities of four beans.

Optimal Liquid Inoculum Conditions and Grain Medium Enhanced Hispidin Production by Species of Genus Phellinus (Agaricomycetes) in Solid-State Fermentation.

This study aimed 1) to screen a high hispidin production strain from 12 strains of genus Phellinus and 2) to evaluate the effects of liquid inoculum conditions and grain medium on this strain's hispidin production levels after solid-state fermentation. The results showed that Ph. linteus 04 led to the highest hispidin production; this strain was then selected to elucidate the optimal liquid inoculum conditions and grain medium for hispidin production. Various liquid inoculum conditions were evaluated, and the highest hispidin yield, specific productivity of hispidin, and total content of hispidin were found to be optimal at 1 week of liquid inoculum culture time, cultured with potato dextrose broth, and using a 10% inoculum rate, with each condition resulting in 0.350, 0.325, and 0.328 mg/g dry weight of mycelium, 0.352, 0.251, and 0.249 µg/mg of specific productivity per week, and 57.90, 60.23, and 61.77 mg/kg dry weight of brown rice medium, respectively. These liquid inoculum conditions were then used to determine the appropriate grain medium for hispidin production. The highest hispidin yield and total content of hispidin were observed in pearl barley (1.107 mg/g dry weight of mycelium and 199.76 mg/kg dry weight of pearl barley), which led to results that were 4.73-fold and 5.35-fold higher than those of the control (brown rice medium). Overall, this study shows that Ph. linteus hispidin production can be enhanced by solid-state fermentation using optimal liquid inoculum conditions and the appropriate grain medium.

Influences of carbon and nitrogen source addition, water content, and initial pH of grain medium on hispidin production of Phellinus linteus by solid-state fermentation.

This study aimed to evaluate the effects of certain grain medium conditions, such as carbon and nitrogen source addition, water content, and initial pH, on the hispidin production of Phellinus linteus using solid-state fermentation. The results showed that the highest hispidin yield, specific productivity of hispidin, and total content of hispidin were associated with sucrose and malt extract addition, 55% water content, and an initial pH of 5.5, which resulted in 0.290, 0.233, 0.301, and 0.296 mg/g dry weight of mycelium; 0.261, 0.191, 0.257, and 0.227 µg/mg week-1 specific productivity; and 46.01, 40.67, 58.85, and 55.06 mg/kg dry weight brown rice medium, respectively. Additionally, two combinations of culture conditions with pearl barley and black rice medium were tested for hispidin production. The highest hispidin yield, specific productivity of hispidin, and total hispidin content for pearl barley medium fermented using P. linteus were 3.88-fold, 4.86-fold, and 3.60-fold higher than those for the control (brown rice medium), respectively. Overall, this study shows that P. linteus hispidin production can be enhanced using solid-state fermentation, with optimal medium conditions.

Biological efficiency and nutritional value of the culinary-medicinal mushroom Auricularia cultivated on a sawdust basal substrate supplement with different proportions of grass plants.

Auricularia polytricha was cultivated on a sawdust basal substrate supplemented with different proportions (30%, 45%, and 60%, respectively) of stalks of three grass plants, i.e., Panicum repens (PRS), Pennisetum purpureum (PPS), and Zea mays (ZMS), to determine the most effective substrate. The mycelial growth rate, total colonization time, days to primordial formation, biological efficiency and chemical composition of fruiting bodies were evaluated. The results indicated that 30PPS was the best substrate for mycelial growth of A. polytricha, with a corresponding total colonization period of 32.0 days. With the exception of 30PPS, the total biological efficiency of all of the substrates containing P. repens stalk, P. purpureum stalk and Z. mays stalk was higher (P < 0.05) than that of the control. The most suitable substrate with a high biological efficiency was 60PRS (148.12%), followed by 30ZMS (145.05%), 45ZMS (144.15%) and 30PRS (136.68%). The nutrient values of fruiting bodies were affected by different substrates. The ash contents of A. polytricha cultivated on a substrate containing Z. mays stalk were higher than that of the control; meanwhile, the protein contents of mushroom cultivated on a substrate containing P. repens stalk (except substrate 45PRS) were higher than that of the control. The biological efficiency of the substrates was tested, and according to the results, it is feasible to use the stalks of P. repens and Z. mays on partially replaced sawdust to cultivate A. polytricha.

Characteristics of Exopolysaccharides from the Citrine Oyster Mushroom, Pleurotus citrinopileatus (Agaricomycetes), Depend on the Nitrogen Source in the Medium.

We investigated the effect of peptone and (NH4)2SO4, used as nitrogen sources in a submerged medium, on the molecular weight and carbohydrate compositions of exopolysaccharides (EPSs) produced by the culinary-medicinal mushroom Pleurotus citrinopileatus. When (NH4)2SO4 was used instead of peptone, it yielded more specific EPSs, producing 32.80 mg EPS/g dry mycelial biomass, but small amounts of EPS and mycelial biomass. These nitrogen sources also resulted in the production of different net charged EPSs, according to crude fractionations on a DEAE Sepharose Fast Flow column. Purified EPSs were initially repurified by gel filtration chromatography, and the profiles indicated that a range of EPSs with different molecular weights were collected through the use of this strategy. Analysis of the carbohydrate composition of purified EPSs demonstrated that the main component produced by P. citrinopileatus in a glucose-(NH4)2SO4 medium, with an estimated molecular weight < 10 kDa, was rich in glucose (95.9%), and its structure could consist of glucan. Conversely, this mushroom produced diverse heteropolysaccharides (11.1-42.3% mannose, 6.3-80.3% glucose, 0.3-49.2% N-acetylgalactosamine, and 7.6-26.3% N-acetylglucosamine) in a glucose-peptone submerged medium. By applying various nitrogen sources in submerged medium, these results could be used to improve the possibility of obtaining molecules similar to those found in the fruiting bodies of the same mushroom species grown in conventional culture.

Effect of Different Proportions of Agrowaste on Cultivation Yield and Nutritional Composition of the Culinary-Medicinal Jelly Mushroom Auricularia polytricha (Higher Basidiomycetes).

In this study, Auricularia polytricha was cultivated on a sawdust basal substrate supplemented with different proportions (30%, 45%, and 60%, respectively) of agrowastes-sugarcane bagasse (SB), rice straw (RS), and rice husk (RH)-to evaluate the alternative substrates. The mycelial growth rate, total colonization time, time to first primordia, biological efficiency, and chemical composition of the fruiting bodies were determined. Results indicated that the 60% SB substrate was the best substrate for mycelial growth of A. polytricha, with a corresponding total colonization period of 35.2 days, followed by the control (35.5 days) and 45% SB (36.2 days) substrates. The most suitable substrate with a high biological efficiency was 60% RS substrate (159.14%), followed by the 45% SB (128.45%), and 20% RH (124.47%) substrates. The nutrient values of fruiting bodies showed the largest amounts of ash, protein, fat, carbohydrates, and energy cultivated on 60% SB, 60% SB, 30% SB, 30% RH, and 30% RH/the control substrates, respectively. The results indicated that 60% RS was an appropriate substrate for A. polytricha cultivation.

Phellinus linteus Mycelium Alleviates Myocardial Ischemia-Reperfusion Injury through Autophagic Regulation.

The incidence of myocardial ischemia-reperfusion (IR) injury is rapidly increasing around the world and this disease is a major contributor to global morbidity and mortality. It is known that regulation of programmed cell death including apoptosis and autophagy reduces the impact of myocardial IR injury. In this study, the cardioprotective effects and underlying mechanisms of Phellinus linteus (Berk. and Curt.) Teng, Hymenochaetaceae (PL), a type of medicinal mushroom, were examined in rats subjected to myocardial IR injury. The left main coronary artery of rats was ligated for 1 h and reperfused for 3 h. The arrhythmia levels were monitored during the entire process and the infarct size was evaluated after myocardial IR injury. Furthermore, the expression levels of proteins in apoptotic and autophagic pathways were observed. Pretreatment with PL mycelium (PLM) significantly reduced ventricular arrhythmia and mortality due to myocardial IR injury. PLM also significantly decreased myocardial infarct size and plasma lactate dehydrogenase level after myocardial IR injury. Moreover, PLM administration resulted in decreased caspase 3 and caspase 9 activation and increased Bcl-2/Bax ratio. Phosphorylation level of AMPK was elevated while mTOR level was reduced. Becline-1 and p62 levels decreased. These findings suggest that PLM is effective in protecting the myocardium against IR injury. The mechanism involves mediation through suppressed pro-apoptotic signaling and regulation of autophagic signaling, including stimulation of AMPK-dependent pathway and inhibition of beclin-1-dependent pathway, resulting in enhancement of protective autophagy and inhibition of excessive autophagy.

Effects of Illumination Pattern during Cultivation of Fruiting Body and Bioactive Compound Production by the Caterpillar Medicinal Mushroom, Cordyceps militaris (Ascomycetes).

We investigated the effects of light intensity in the 3 cultivation stages separately-the mycelium colonization stage, the primordial initiation stage, and the fruiting stage (in order)-on fruiting body and bioactive compound production by Cordyceps militaris. In the mycelium colonization stage, rice substrates were incubated in a spawn running room at 23°C. During the primordial initiation stage, C. militaris was grown at 18°C and illuminated 12 hours/day. In the fruiting stage the temperature was 23°C, with illumination provided 12 hours/day. The highest fruiting body yield and biological efficiency were 4.06 g dry weight/bottle and 86.83%, respectively, under 1750 ± 250 lux during the second and third stages. The cordycepin content was highest during the second and third stages under 1250 ± 250 lux. The mannitol and polysaccharide contents were highest under 1250 ± 250 and 1750 ± 250 lux during the primordial initiation stage and the fruiting stage, respectively. Thus, with controlled lighting, C. militaris can be cultivated in rice-water medium to increase fruiting body yield and bioactive compound production.

Various grain substrates for the production of fruiting bodies and bioactive compounds of the medicinal caterpillar mushroom, Cordyceps militaris (Ascomycetes).

In this study, several grains such as brown rice (Br), plumule rice (Pr), wheat (W) and pearl barley (Pb) supplemented with 1% (w/w) peptone (P), yeast extract (Ye), ammonia sulfate (As), and monosodium glutamate (Mg) as a nitrogen source, respectively, were used to produce fruiting bodies and bioactive compounds of two strains of Cordyceps militaris. Among these grain substrates, the substrate most suitable to mycelial growth was Pb+Ye for C. militaris H and L. The mushroom strains colonized this substrate in 12.8 and 12.6 days, respectively. For C. militaris L, the fewest days were required for primordial initiation on Br+Ye and Pr+P substrates. The highest yield and biological efficiency was observed with Pb substrate (25.16 g/bottle and 87.36%) and Br+P substrate (21.84 g/bottle and 75.83%) for C. militaris H and L, respectively. In the fruiting bodies of C. militaris H, the highest cordycepin content was cultivated on W+Mg substrate (25.07 mg/g), the highest mannitol content was cultivated with Pr+Mg (153.21 mg/g) and Pr (151.65 mg/g) substrates, and the highest adenosine content was cultivated with Pr+Ye (0.94 mg/g) and Pb+Ye (0.90 mg/g) substrates. In the fruiting bodies of C. militaris L, the highest cordycepin content was cultivated with W+Mg substrate (22.14 mg/g); the highest mannitol content was cultivated with Pb substrate (189.33 mg/g); and the highest adenosine content was cultivated with Pb+Ye substrate (0.71 mg/g).

Effect of light and atmosphere on the cultivation of the golden oyster culinary-medicinal mushroom, Pleurotus citrinopileatus (higher Basidiomycetes).

With an aim to explore the productivity and quality of the fruiting body of culinary-medicinal golden oyster mushroom Pleurotus citrinopileatus, the carbon dioxide (CO2) concentration of the ambient atmosphere was adjusted and a light-emitting diode panel was used to illuminate the colonized mycelium at different wavelengths. Biological efficiency and yield were higher at CO2 levels of 0.05 and 0.1% than other tested CO2 levels, and the mature fruiting body showed the highest yellow value at a CO2 level of 0.1% (of all tested CO2 levels). The highest biological efficiency and yield was obtained at the 720-nm wavelength. The ergosterol content of the pileus of the fruiting body was higher than that of the stipe in any flush time at a 720-nm wavelength of light and a CO2 concentration of 0.1%. The decreased percentages of cellulose and lignin at the appearance of primordia were larger than those of mycelial growth duration. The fruiting quality of P. citrinopileatus might thus be enhanced by 720-nm illumination and an atmosphere with a CO2 concentration of 0.1 to 0.15%.

Cultivation of the culinary-medicinal Lung Oyster mushroom, Pleurotus pulmonarius (Fr.) Quél. (Agaricomycetideae) on grass plants in Taiwan.

Cultivation of the culinary-medicinal Lung Oyster mushroom, Pleurotus pulmonarius, on the stalks of three grass plants, i.e., Panicum repens, Pennisetum purpureum, and Zea mays were investigated. The effects of various combinatorial substrates on mushroom mycelial growth and yield calculated as biological efficiency (BE) were determined. Among 9 experimental substrates, the most suitable substrate for mycelial growth was 45ZMS:45S, followed by 45PRS:45S; their mycelial growth rates were obviously quicker than that of the control substrate. The BEs of all the experimental substrates respectively containing P. repens stalk, P. purpureum stalk and Z. mays stalk were higher than that of the control (39.55%) during the 2.5 months of cultivation period. The best substrate in terms of BE was 60ZMS:30S (58.33%), followed by 45PRS:45S (57.16%), 45ZMS:45S (49.86%), and 30ZMS:60S (47.20%). Based on the BE of the tested substrates, Z mays stalk appeared to be the best alternative material for the production of P. pulmonarius.