Submerged culture conditions for mycelial yield and polysaccharides production by Lyophyllum decastes

The effects of various carbon and nitrogen sources, their concentrations, initial pH and fermentation duration on the production of mycelia in terms of dry weight, exo-polysaccharide (EPS) and inner polysaccharide (IPS) by Lyophyllum decastes, a culinary-medicinal mushroom, were investigated in shake-flask cultures. Lactose, glucose and fructose were the top three best carbon sources for mycelial growth with corresponding yields of 6.73 g/l, 6.36 g/l and 6.10 g/l, respectively. Glucose was the best for production of EPS and IPS with 1.65 g/l and 317 mg/g dry mycelia, respectively. Maltose also performed well for EPS production. Yeast extract was the best nitrogen source for the production of mycelia (7.03 g/l) and IPS (325 mg/g dry mycelia), whereas EPS was improved further by increasing the yeast extract concentration (2.46 g/l at 2%). Similarly, initial pH 7 and 8 were best for polysaccharides production (EPS 1.73 g/l and IPS 320 mg/g) and mycelial growth (7.10 g/l), respectively. Maximum mycelial growth peaked at 15 days of cultivation whereas polysaccharides peaked at 10 days, and then tapered off. A concentration of glucose 3% and yeast extract 1% (mycelial yield and IPS) were found to be a suitable condition for submerged culture.     

Short communication: Cultivation of Lentinus edodes mycelia using whey permeate as an alternative growth substrate

The major objective of this research was to use whey permeate as an alternative growth medium for the cultivation of mycelia of the edible mushroom Lentinus edodes and to find an optimum condition for solid-state cultivation. Response surface analysis was applied to determine the combination of substrate concentration (40 to 60 g of lactose/L), temperature (20 to 30°C), and pH (4 to 6) resulting in a maximal mycelial growth rate. The radial extension rates, estimated by measuring the diameters of growing colonies on the Petri dishes, were used as the growth rate of the mycelia at different conditions. The conditions predicted to maximize the mycelial growth of 6.41 ± 0.47 mm/d were determined to be 40 g of lactose/L, temperature 23.6°C, and pH 5.0. It was concluded that a partial cubic equation could accurately model the response surface of, and predict optimal growth conditions for, L. edodes mycelia using whey permeate because the model prediction agreed with the experimental growth rate, 6.39 ± 0.22 mm/d. The results suggest that whey permeate could be utilized as a growth substrate for the cultivation of mycelia from the edible mushroom L. edodes, enhancing the use of this by-product by the cheese manufacturing industry.     

Optimization of Selenium-Enriched Mycelium of Lentinula edodes (Berk.) Pegler as a Food Supplement

Our goal was to optimize the growth conditions of submerged mycelial cultures of Lentinula edodes (Shiitake mushroom) in order to obtain a new dietary supplement enriched in selenium We designed a process technology in which mycelial cultures were cultivated in media composed of beet molasses, 10%; liquid stillage, 5%; corn steep liquor, 0.15%; and KH₂PO₄, 0.3%, enriched with selenium in concentrations ranging from 0 to 100 μg/mL by the addition of sodium selenite of selenic acid. Se concentrations in mycelial dry mass rose from 0.001 mg/g (mycelia cultivated in media containing no selenium) to 50 mg/g (mycelia cultivated in medium containing 100 μg Se/mL). The highest mycelial specific growth rate (0.46/day) was recorded when the concentration of selenium in the medium was lower than 20 μg/mL. In vitro, estimated Se bioavailabilities from selenized mycelium strongly depended on its preparation in a proper manner and were 60% and 82% for dried mycelium and mycelial lyophilizate, respectively. Our results suggest that these optimized culture conditions could be applied to obtain a new Se-enriched dietary supplement.     

Use of whey permeate for cultivating Ganoderma lucidum mycelia

A novel approach to utilizing whey permeate, the cultivation of mycelia of the edible mushroom Ganoderma lucidum, is introduced. The major objective of this research was to use whey permeate as an alternative growth medium for the cultivation of mycelia of edible mushroom G. lucidum and to find an optimum condition for solid-state cultivation. Response surface analysis was applied to determine the combination of substrate concentration (25 to 45 g of lactose/L), pH (3.5 to 5.5), and temperature (25 to 35°C) resulting in a maximal mycelial growth. The radial extension rates, estimated by measuring the diameters of growing colonies on the Petri dishes, were used as the growth of the mycelia at different conditions. In the model, pH and temperature significantly affected mycelial growth, but lactose concentration did not. The condition predicted to maximize the radial extension rate of 17.6 ± 0.4 mm/d was determined to be pH 4.4 and temperature 29.4°C. Therefore, the results suggest that whey permeate could be utilized as a growth substrate for the cultivation of mycelia from the edible mushroom G. lucidum, enhancing the use of this by-product by the cheese manufacturing industry.     

Nonvolatile taste components of Grifola frondosa, Morchella esculenta and Termitomyces albuminosus mycelia

Three species of mushroom mycelia are commercially available in Taiwan, namely Grifola frondosa (maitake), Morchella esculenta (morel), and Termitomyces albuminosus (termite mushroom), and their nonvolatile taste components were studied. All mycelia were high in contents of carbohydrate, crude fat and protein but low in contents of crude ash and fiber. Arabitol, glucose, mannitol and trehalose were detected in these three mycelia and total amounts of sugars and polyols were 63.69-83.89 mg/g. Total free amino acid contents in three mycelia ranged from 35.67 to 50.37 mg/g. The monosodium glutamate-like components of G. frondosa mycelia (6.51 mg/g) was approximately twofold higher than M. esculenta and T. albuminosus mycelia (3.58 and 3.11 mg/g, respectively). As compared with other components, sweet and bitter components were much higher in these three mycelia. Contents of total 5′-nucleotides in these three mycelia ranged from 13.32 to 26.19 mg/g and were in the descending order of T. albuminosus>M. esculenta>G. frondosa. Equivalent umami concentration of T. albuminosus mycelia was higher than those of G. frondosa and M. esculenta mycelia. Overall, three mycelia possessed highly intense umami taste.     

Selenium contents of Portuguese commercial and wild edible mushrooms

The selenium contents of 66 mushroom samples, wild growing or commercially available in Portugal, fresh, canned or dried, were determined. The samples consisted of 18 mushroom species, mainly selected in accordance with their availability and consumer preference. The selenium contents varied considerably between different mushroom species. The highest selenium contents were found in the wild species Boletus aestivalis (48.5 mg/kg dry weight, DW), Boletus pinophilus (19.9 mg/kg DW), Boletus edulis (14.9 mg/kg DW), Boletus aereus (12.3 mg/kg DW), Boletus fragans and Boletus spretus (∼2 mg/kg DW). Other mushrooms having considerable selenium contents included Marasmius oreades (1.5 mg/kg DW), Agaricus bisporus “Portobello” (1.25 mg/kg DW), A. bisporus (1.0 mg/kg DW) and Russula cyanoxantha (0.72 mg/kg DW). The selenium contents of these mushrooms are sufficient to provide nutritionally significant amounts in relation to the total daily intake of selenium. Other edible mushrooms, such as Lentinula edodes, Pleurotus ostreatus, Cantharellus cibarius,Craterellus cornucopioides and Lepista nuda, contained only small amounts of selenium. The importance of these mushrooms as a source of selenium is therefore marginal.     

Antioxidant activity and phenolic content of 16 raisin grape (Vitis vinifera L.) cultivars and selections

Six raisin grape cultivars and 10 new raisin grape selections were analyzed for antioxidant activity (ABTS assay) and for total and individual phenolic compounds. Samples were freeze–dried and values are reported on a dry weight basis. Antioxidant activity across the 16 samples ranged from 7.7 to 60.9 μmol Trolox/g DW, with A95-27 exhibiting the greatest activity. Total phenolic content, determined in gallic acid equivalents using the Folin–Ciocalteau assay, ranged from 316.3 to 1141.3 mg gallic acid/100 g DW and was strongly correlated (r = 0.990) with antioxidant results. Concentrations of individual phenolics were determined by HPLC. trans-Caftaric acid was the predominant compound in all samples. A95-15 contained the lowest concentration (153.5 μg/g DW) of caftaric acid, while Fiesta contained the highest concentration (598.7 μg/g DW). Selections A56-66, A95-15, and A95-27 had much higher levels of catechin (86.5–209.1 μg/g DW) and epicatechin (126.5–365.7 μg/g DW) than the other samples.     

Chemical,biochemical and electrochemical assays to evaluate phytochemicals and antioxidant activity of wild plants

Plants are a source of compounds that may be used as pharmacologically active products. Cytisus multiflorus, Filipendula ulmaria and Sambucus nigra have been used as important medicinal plants in the Iberian Peninsula for many years, and are claimed to have various health benefits. Herein, the phytochemical composition and antioxidant activity of the mentioned wild medicinal plants were evaluated in vitro, based on chemical, biochemical and electrochemical methods. F. ulmaria was found to be richest in antioxidant phytochemicals, such as phenolics (228 mg GAE/g DW), flavonoids (62 mg CE/g DW), ascorbic acid (2700 μg/g DW) and tocopherols (497 μg/g DW). The antioxidant activity was found to vary in the order: F. ulmaria > S. nigra > C. multiflorus, irrespective of the analysis method. Electrochemical methods have proven to be rapid and inexpensive techniques to characterise the antioxidant activity of plant extracts.     

Composition and non-volatile taste components of Hypsizigus marmoreus

Two strains of Hypsizigus marmoreus (Peck.) Bigelow (Tricholomataceae) are successfully cultivated and commercially available in Taiwan, and their composition and non-volatile taste components of fruit bodies and mycelia were studied. Both fruit bodies were higher than mycelia in contents of carbohydrate, ash and fiber but lower in contents of fat and protein. Total sugar and polyol contents were 45.47-91.50 mg/g and total free amino acid contents were in the descending order of white strain fruit bodies (122.97) > normal strain fruit bodies (95.94) > white strain mycelia (53.20) > normal strain mycelia (46.87 mg/g). Monosodium glutamate-like components of both fruit bodies were 3-4-fold higher than those of both mycelia. Total 5′-nucleotides contents were 6.43-11.02 mg/g with white strain fruit bodies being the highest. Equivalent umami concentrations of both fruit bodies were higher than those of mycelia. Overall, H. marmoreus fruit bodies possessed highly intense umami taste.     

Chemical characterisation and speciation of organic selenium in cultivated selenium-enriched Agaricus bisporus

The selenium concentration in Agaricus bisporus cultivated in growth compost irrigated with sodium selenite solution increased by 28- and 43-fold compared to the control mushroom irrigated solely with water. Selenium contents of mushroom proteins increased from 13.8 to 60.1 and 14.1 to 137 μg Se/g in caps and stalks from control and selenised mushrooms, respectively. Selenocystine (SeCys; detected as [SeCys]₂ dimer), selenomethionine (SeMet), and methyl-selenocysteine (MeSeCys) were separated, identified and quantified by liquid chromatography–electrospray ionisation-mass spectrometry from water solubilised and acetone precipitated proteins, and significant increases were observed for the selenised mushrooms. The maximum selenoamino acids concentration in caps and stalks of control/selenised mushrooms was 4.16/9.65 μg/g dried weight (DW) for SeCys, 0.08/0.58 μg/g DW for SeMet, and 0.031/0.10 μg/g DW for MeSeCys, respectively. The most notable result was the much higher levels of SeCys accumulated by A. bisporus compared to SeMet and MeSeCys, for both control and selenised A. bisporus.     

Nonvolatile taste components of fruit bodies and mycelia of Cordyceps militaris

Cordyceps militaris (L.) Link (Clavicipitaceae), northern Cordyceps, is currently available in Taiwan for use in the formulation of nutraceuticals and functional foods. The nonvolatile components in the form of fruit bodies and mycelia were studied. Both fruit bodies and mycelia were high in contents of carbohydrate, crude fiber and crude protein. Content of total sugars and polyols were 260.64 and 189.82 mg/g for fruit bodies and mycelia, respectively. Mannitol content was the highest in both fruit bodies and mycelia (117.66 and 112.55 mg/g, respectively). Contents of total free amino acids in fruit bodies and mycelia were 48.15 and 67.63 mg/g, respectively. The content of monosodium glutamate (MSG)-like components in fruit bodies (10.60 mg/g) was higher than that in mycelia (2.70 mg/g). The contents of total and flavor 5′-nucleotides were high in mycelia (26.27 and 9.34 mg/g, respectively). Equivalent umami concentrations of fruit bodies and mycelia were similar, and equivalent to the umami intensity given by 6.08 and 6.22 g of MSG, respectively. Overall, both fruit bodies and mycelia of northern Cordyceps possessed highly intense umami taste.     

Comparison of the antioxidant activity of two Spanish onion varieties

Phenol content and antioxidant activity of two Spanish onion varieties, namely white onion and Calçot de Valls, have been studied. White onions contained higher phenol content than Calçot onions, with values which ranged from 2.57 ± 0.51 to 6.53 ± 0.16 mg gallic acid equivalents/g dry weight (GAE/g DW) and 0.51 ± 0.22 to 2.58 ± 0.16 mg GAE/g DW, respectively, depending on the solvent used. Higher phenol content was associated with higher antioxidant capacity. White onion extracts had the highest antioxidant activity at 86.6 ± 2.97 and 29.9 ± 2.49 μmol Trolox/g DW for TEAC and FRAP assays, respectively, while the values for the Calçot variety were 17.5 ± 0.46 and 16.1 ± 0.10 μmol Trolox/g DW.     

Isolation and quantitative determination of ergosterol peroxide in various edible mushroom species

Ergosterol peroxide, the steroidal derivative with cytotoxic activity, has been isolated for the first time from the mycelium of edible and medicinal mushroom Hericiumerinaceum (lion’s mane mushroom) together with erinacine A. The new densitometric method was applied for the quantitative determination of ergosterol peroxide in n-hexane extracts of H. erinaceum, Laetiporus sulfureus (chicken mushroom), and Morchella esculenta (common morel) mycelia, as well as in Boletus edulis (king bolete), Suillus bovinus (Jersey cow mushroom), and B. badius (bay bolete) fruiting bodies. The ergosterol peroxide content reached 15.98 ± 0.78, 10.07 ± 0.75, 13.37 ± 0.56, 29.32 ± 1.43, 17.27 ± 0.84, and 12.60 ± 0.59 mg per 100 g, respectively. What is significant was that ergosterol peroxide was identified for the first time, to the best of our knowledge, in edible mushrooms mentioned above.     

Effect of nutritional and environmental conditions on the production of exo-polysaccharide of Agaricus brasiliensis by submerged fermentation and its antitumor activity

Mushrooms have become attractive as a functional food and as a source for the development of drugs and nutraceuticals. A. brasiliensis is considered as the best among them. There are several published works on the fruiting bodies of this mushroom, showing high antitumor activity. The mycelium polysaccharide and exo-polysaccharide (EPS) of this mushroom also demonstrated a strong antitumor action. However, there is little information available in the literature about the optimization of physical and chemical conditions for production of EPS by submerged fermentation (SmF). After initial screening from the five strains available in the LPB Culture Collection, A. brasiliensis LPB 03 was selected for EPS production in SmF. Studies on supplementation of a carbon source to the medium revealed that sucrose was most effective for EPS production. Yeast extract was the best for EPS among the inorganic and organic nitrogen sources tested. The factorial experiment demonstrated that a temperature of 30 °C and a pH of 6.1 were the best for the EPS production. Glucose 10 g/l, yeast extract 3 g/l, K₂HPO₄ 0.6 g/l and MgSO₄ 0.3 g/l were most suitable for the EPS production. Maximum EPS production was obtained in the static condition with manual agitation two times per day after a 1-week culture (382 mg/l), increasing by 78.62% more than that before optimization. In the bioreactor with aeration and agitation, the maximum EPS was produced at 321.2 mg/l. The EPS of A. brasiliensis produced in SmF showed strong inhibition against Sarcoma 180 in mice, reaching 72.19% inhibition compared with the control group. Furthermore, 50% of mice in the test group demonstrated total tumor regression.     

Antioxidant properties of methanolic extracts from Agrocybe cylindracea

Agrocybe cylindracea (DC: Fr.) Mre. (Bolbitiaceae) was available in the form of fruit bodies, mycelia and fermentation filtrate. From these three forms, methanolic extracts were prepared and their antioxidant properties studied. The methanolic extract from fruit bodies showed high inhibitory ability on lipid oxidation (90.0-97.3%) at 5-20 mg/ml and had an EC₅₀ value of 0.94 mg extract/ml. EC₅₀ values in reducing power were 2.36, 6.89 and 5.00 mg extract/ml for fruit bodies, mycelia and filtrate, respectively. The scavenging ability of the methanolic extract from fruit bodies on 1,1-diphenyl-2-picrylhydrazyl radicals was 89.0% at 1 mg/ml whereas those from mycelia and filtrate was 91.4% and 94.9% at 10 mg/ml, respectively. The chelating abilities of methanolic extracts from fruit bodies, mycelia and filtrate on ferrous ions were 90.6%, 84.6% and 96.3% at 5 mg/ml, respectively. Total phenols were the major detected naturally occurring antioxidant components found in methanolic extracts from A. cylindracea and in the range of 15.55-23.47 mg/g. Based on EC₅₀ values, A. cylindracea was good in antioxidant properties, except for the scavenging ability on hydroxyl radicals.     

Identification and quantification of major phenolic compounds from mango (Mangifera indica, cv. Ataulfo) fruit by HPLC–DAD–MS/MS-ESI and their individual contribution to the antioxidant activity during ripening

Mango (Mangifera indica L.) is an economically important fruit throughout the world. ‘Ataulfo’ mango, a leading cultivar in Mexico, has the highest content of phenolic compounds among several commercial varieties of mango. However, the individual identification and antioxidant contribution of these phenols during ripening of mango fruit is unknown. Qualitative and quantitative analysis of the major phenolic compounds found in ‘Ataulfo’ mango fruit pulp was conducted in four stages of ripeness, using high-performance liquid chromatography coupled to mass spectrometry. The antioxidant contribution of each of the major phenolic compounds was calculated. The major compounds identified were chlorogenic acid (28–301 mg/100 g DW), gallic acid (94.6–98.7 mg/100 g DW), vanillic acid (16.9–24.4 mg/100 g DW), and protocatechuic acid (0.48–1.1 mg/100 g DW). The antioxidant contribution of the four phenolic acids increased during ripening. Gallic acid accounted for the highest contribution (39% maximum value), followed by chlorogenic acid (21% maximum value). This could indicate that these phenolic compounds may have an important role in the antioxidant metabolism in ‘Ataulfo’ mango fruit during ripening, and promoting health benefits to consumers.     

Chemical composition of fruits in some rose (Rosa spp.) species

Fruits of Rosa canina, Rosa dumalis subsp. boissieri, Rosa dumalis subsp. antalyensis, Rosa villosa, Rosa pulverulenta and Rosa pisiformis were assayed for total phenolics, ascorbic acid, total soluble solids, total dry weight, total fat, fatty acids, pH, acidity, moisture, fruit colour and macro- and micro-elements. The highest total phenolic content was observed in Rosa canina (96 mg GAE/g DW). Rosa dumalis subsp. boissieri had the highest total fat content (1.85%), followed by Rosa pulverulenta (1.81%) and Rosa canina (1.78%), respectively. Nine major fatty acids were determined in rose species and α-linolenic acid was found to be dominant for all species. Total soluble solids, total dry weight, moisture and ascorbic acid contents of rose species varied from 29.42% (Rosa villosa)–37.33% (Rosa dumalis subsp. boissieri), 33.85% (Rosa villosa)–40.35% (Rosa dumalis subsp. boissieri), 59.65% (Rosa dumalis subsp. boissieri)–66.15% (Rosa villosa) and 727 mg/100 g FW (Rosa villosa) and 943 mg/100 g FW (Rosa dumalis subsp. boissieri), respectively. Nitrogen and mineral compositions of the rose species, e.g., N, P, K, Ca and Mg, were (averagely): 1.26%, 513 mg/100 g DW, 639 mg/100 g DW, 196 mg/100 g DW and 114 mg/100 g DW, respectively. The present study shows that the native rose genotypes are extremely rich sources of phenolics, carbohydrates and ascorbic acid, demonstrating their potential use as a food or food additive.     

Effects of binary solvent extraction system,extraction time and extraction temperature on phenolic antioxidants and antioxidant capacity from mengkudu (Morinda citrifolia)

An investigation into the effects of ethanol concentration (0–100%, v/v), extraction time (20–120 min) and extraction temperature (25–65 °C) on the extraction of phenolic antioxidants from mengkudu (Morinda citrifolia) was performed using a single-factor experiment. Total phenolic content (TPC) and total flavonoid content (TFC) assays were used for determination of phenolic compounds. Antioxidant capacity was evaluated by measuring the scavenging effect on 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radicals. Experimental results showed that extraction conditions had significant effect on extraction of phenolic compounds and antioxidant capacities. The optimised conditions were 40% ethanol for 80 min at 65 °C, with values of 919.95 mg GAE/100 g DW for TPC, 472.73 mg CE/100 g DW for TFC, 791.71 μmol TEAC/100 g DW for ABTS and 1928.5 μmol TEAC/100 g DW for DPPH. TPC was significantly correlated with DPPH under the effects of ethanol concentration (r = 0.932) and extraction time (r = −0.938).     

UPLC and HPLC of caffeoyl esters in wild and cultivated Arctium lappa L.

Analytical methods including ultra-performance liquid chromatography (UPLC) and high-performance liquid chromatography (HPLC) with photodiode array (PDA) detector were developed for the analysis of caffeoylquinic acid derivatives in seeds, leaves and roots of Arctium lappa L. Separation was performed on C₁₈ column utilising 5% (v/v) acetic acid in water and acetonitrile at 330 nm. Both methodologies were validated in terms of linearity, precision, and recovery. The results showed that the major advantages of UPLC, over HPLC were the fast analysis, narrow peaks, high sensitivity, and reduction of solvent consumption. Subsequently the methods were applied for the identification and quantification of chlorogenic acid (5-CQA) and 1,5-dicaffeoylquinic acid (1,5-DCQA) as main compounds in samples. The total phenolic content of samples ranged from 3.93 to 14.13 g of 5-CQA equivalent/100 g dry weight (DW). There was a significant variability from 89 to 571 mg/100 g for 5-CQA and 48 to 486 mg/100 g for 1,5-DCQA in dry material.     

Nutrient composition of hazelnut (Corylus avellana L.) varieties cultivated in Turkey

In this study, chemical composition of the 17 different hazelnut varieties grown in the Black Sea Region of Turkey was investigated. The main fatty acids in hazelnut varieties were oleic (79.4%), linoleic (13.0%) and palmitic acid (5.4%). The ratios of polyunsaturated/saturated and unsaturated/saturated fatty acids of hazelnuts varieties were found to be between 1.23 and 2.87, and 11.1 and 16.4, respectively. The average niacin, vitamin B₁, vitamin B₂, vitamin B₆, ascorbic acid, folic acid, retinol and total tocopherol contents of hazelnut kernels were 1.45 mg/100 g, 0.28 mg/100 g, 0.05 mg/100 g, 0.5 mg/100 g, 2.45 mg/100 g, 0.043 mg/100 g, 3.25 mg/100 g and 26.9 mg/100 g, respectively. The amount of the essential amino acids, mostly as arginine (2003 mg/100 g) and leucine (1150 mg/100 g), and the non-essential amino acids, mostly as glutamic acid (2714 mg/100 g) and aspartic acid (1493 mg/100 g) were also determined in the hazelnut varieties. Mineral compositions of the hazelnut varieties, e.g., K, Mn, Mg, Ca, Fe, Zn, Na and Cu were (averagely) measured as 863 mg/100 g, 186 mg/100 g, 173 mg/100 g, 5.6 mg/100 g, 4.2 mg/100 g, 2.9 mg/100 g, 2.6 mg/100 g and 2.3 mg/100 g, respectively.