Nutrient composition and functional activity of different stages in the fruiting body development of Philippine paddy straw mushroom, volvariella volvacea (Bull.:Fr.) sing.
Edible mushrooms have been used as source of therapeutic agents because of their distinctive chemical diversity and promising healthful benefits since time immemorial. Bioactive compounds derived from mushrooms include polysaccharides, proteins, protein-polysaccharide complex, lipids and other metabolites such as heterocyclics, polyketides, sterols, terpenes, peptides, terphenyls [1, 2, 3, 4, 5, 6]. There are wide varieties of chemicals in mushrooms like beta-glucan, a polysaccharide that exhibits numerous functional activities. Beta glucans are considered to be the most potent bioactive fungal compounds [7, 8] which show unique character in individual basidiomycetes. Previously, schizophyllan from Schizophyllum commune, grifolan from Grifola frondosa, and lentinan from Lentinula edodes [9, 10, 11] have been comprehensively studied for their therapeutic properties.
Another important functional fungal component is the bioactive proteins. Lectins, ribosome inactivating proteins, antimicrobial proteins, immunomodulatory proteins, ribonucleases, laccases, chitinases, defensins, glucanases, lipid transfer proteins, protease inhibitors, peroxidases and other proteins have been reported to demonstrate medicinal activities [12, 13]. Apart from these diverse proteins, amino acids of edible mushrooms were also extensively studied by many researchers [14, 15, 16, 17]. In fact, our research team has established the amino acid profile and functional activities of some Philippine native and exotic species of edible mushrooms namely: Schizophyllum commune, Lentinus tigrinus, Lentinus sajor caju, Ganoderma lucidum, Pleurotus florida, Coprinus comatus and Collybia reinakeana [18,19,20]. Recently, it was reported the bioactive metabolites from mushrooms that exhibit anti-diabetic, anti-malarial, anti-microbial, anti-oxidant, anti-Alzheimer, antitumor, anti-viral and hypocholesterolemic activities . Some of these novel bioactive metabolites include styrylpyrones from Inonotus and Phellinus species, cyathusals and pulvinatal from Cyathus stercoreus, triterpenoids from Ganoderma lucidum, cordycepin from Cordyceps species and among others [22, 23, 24, 25].
Volvariella volvacea, the paddy straw mushroom, is the most popular edible mushroom for Filipinos because this is the first mushroom species introduced in the Philippines . It is vernacularly termed as kabuteng dayami or kabuteng saging when found growing on decomposing piles of rice straw and decaying banana leaves, respectively. This mushroom has grayish to brown pileus and white to brownish silky stipe encased in brownish gray sack-like volva, egg-shaped when young and expanding broadly conic to nearly flat pileus when mature. In the past, fruiting bodies of this tropical mushroom were usually collected by the farmers from the wild during the onset of rainy seasons. However, due to the successful development of production technologies, V. volvacea can be grown throughout the year. The fruiting bodies can be harvested 14 days after spawning of the substrate with 65% moisture content, incubated at 30-35.C with very minimal ventilation and profise lighted conditions . Interestingly, there are reports referring to V. volvacea as an ideal source of antimicrobial, anti-cancer, antioxidants, anti-tumor substances [28, 29, 30] and nutritive values such as crude protein, crude fibre, carbohydrates, ascorbic acid and minerals like potassium, phosphorous, magnesium, calcium, zinc, iron, manganese, and copper .
This work investigated the nutraceutical attributes and in vitro biological activities of different stages growth of fruiting body of Volvariella volvacea to establish its important role in nutraceutical and pharmaceutical industiy. These benchmark information obtained in the present study would contribute to the recent advances in the field of medicine and developments novel therapies to fight coagulation, inflammation and hypertension and their related complications.
MATERIALS AND METHODS
Cultivation of Mushroom Samples:
Paddy straw mushrooms were cultured as follows. 20 kgs rice straw containing 2% wheat bran, 1% ammonium sulfate, 1% lime and 0.5% urea was fermented for 14 days, turned, and adjusted to pH 6.0 with 65% moisture. The formulated substrate was steam- sterilized at 121[degrees]C, 15 psi for 40 minutes. Upon cooling, the formulated substrate was inoculated with spawn of V volvacea and subsequently incubated at 30-35.C for 10-14 days to allow mycelial ramification and subsequently the emergence of fruiting bodies. Fruiting bodies (Figure 1) were harvested in each of the four growth stages (I, egg-shaped; II, ruptured of the volva; III, emergence of the pileus; IV, fully expanded pileus).
Preparation of Mushroom Samples for Analysis:
Immediately after harvest, fruiting bodies were freeze-dried to be used for component analysis and functionality evaluation tests.
Nutrient Component Analysis:
Amino acids, polysaccharides, organic acids, and minerals were analyzed following the standard protocol for chemical analysis . The amino acid profiles  were elucidated using L-8900 amino acid analyzer (Hitachi Co. Ltd. Japan).
The following assays were performed: inhibition of platelet aggregation, inhibition of chemokine gene expression, and angiotensin converting enzyme (ACE) inhibition activity.
The determination of anti-platelet aggregation induced by platelet activating factor (PAF) and arachidonic acid Na (AA) was conducted by collecting human peripheral blood from the median cubital vein of a medication- free healthy adult for at least 2 weeks. The collected blood was centrifuged at 1100 rpm for 20 min under ambient room temperature. After centrifirgation, the platelet rich plasma (PRP) located at the upper layer was collected. The platelet poor plasma (PPP) located on the lower layer on the other hand was centrifuged at 3000 rpm for 5 min at ambient room temperature. Both plasma (PRP and PPP at 223 [micro]L each were preheated at 37[degrees]C. The methanol extract of each sample of the mushroom extract was dissolved separately in 2 [micro]l of a 2% dimethylsulfoxide (DMSO) solution, and added to PRP and PPP. The set--ups were incubated for 3 min at 37 [degrees]C. Subsequently, in order to induce platelet aggregation, 25 [micro]L of an aqueous solution of 500 nM arachidonic acid (PAF) was added and induced aggregation was measured using an aggregometer (MCM Hema Tracer 313M, MC Medical Co., Ltd. Tokyo, Japan). Ion exchanged water served as control. The inhibitory effect of the mushroom extract was evaluated by comparing the maximum aggregation rates (i.e. maximum value required from aggregation curve of the test sample extract in normalizing the value of the PPP sample to 100) with the control. Following the protocol of Morimitsu  the inhibition rates were normalized to control for calculation of the efficacy of the test sample extracts.
The suppression of chemokine gene expression was determined as follows: human skin fibroblasts were cultivated in Dulbecco's modification Eagle's medium (DMEM) with 10% fetal bovine serum until the confluent growth was attained at 6 cm in diameter. The methanol extract of the sample was placed in the dish at a final dry mass concentration of 0.01%. Hydrocortisone at [10.sup.-7] M served as positive control. Tumor necrosis factor (TOT-alpha) which promotes chemokine gene expression at 1 ng m[L.sup.-1] was added and subsequently incubated for 6 h at 37[degrees]C. Similarly, a TNF--alpha free sample was also incubated in the same conditions for the sample free medium as a control. Based on manufacturer's instruction, total RNA was isolated using ISOGEN reagent (Nippon Gene Co., Ltd., Tokyo, Japan). Total RNA (1 [micro]g) was reverse transcribed to cDNA with M-MEV reverse transcriptase (Life Technologies Co. Ltd., Rockllle, USA) following the product manual. Quantitative real time RT-PCR method was used in measuring IL-8 gene expression. Using TaqMan reverse transcription reagents and the ABI Prism 7700 sequence detection system (Applied Biosystems, Foster City, CA), cDNA was prepared and the samples were quantified using TaqMan universal PCR master mix. cDNA was synthesized by TaqMan RT (AP Bio). The nucleotide sequences for PCR primers and probes were as follows: IL-8 forward primer, 5'- TCAGAGACAGCAGAGCACACA-3'; reverse primer, 5' CTTGGCAGCCTTCCTGATT-3'; probe, 5'- AACATGACTTCCAAGCTGGCCA-3'; GAPDH forward primer, 5'-GAAGGTGAAGGTCGGAGTC-3'; reverse primer, 5'-GAAGATGGTGATGGGATTTC-3'; probe, 5'-AGGCTGAGAACGGGAAGCTTG-3.
The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal standard gene. For the calculation of the efficacy of the test samples and following the works of Leutenegger et al. , the inhibition rates were normalized to that of TNF-alpha. Data were expressed as mean and standard deviation of five replicates each treatment.
The assay on the inhibition of angiotensin converting enzyme (ACE) was adopted from the methods of Reyes et al  with minor modification as follows: 5% concentration of the test sample (i.e. 5 mg of mushroom hot water extract was mixed in 1 ml 100 mM Borate buffer, pH 8.3) was prepared and subsequently filtered through a 0.45 um nylon syringe filter (Whatman, Inc. USA). Ten [micro]l of the sample was mixed in 20 [micro]l 60 mU/ml ACE (from rabbit lung) and 30 [micro]l 1M NaCl and pre incubated at 37[degrees]C for 5 minutes in a water bath. After pre incubation, 60 [micro]l 6mM Hippuric acid-Histidine-Leucine (Hip-His-Leu) which served as the substrate of the enzyme was added to the mixture and incubated in water bath at 37[degrees]C for 60 minutes. The reaction was terminated by adding 60 [micro]l IN HCl. Using a 0.45 um nylon syringe (Millipore Millex[R], LH, Japan), the supernatant was filtered. The filtrate was subjected to reversed phase HPLC with the following conditions: injected dose, 20 [micro]l 1V; analytical column, C18 (GL Sciences, Inc. Japan); Mobile phase, methanol:10mM KH2PO4 at 1:1 and adjusted to pH 3.0 using phosphoric acid; flow rate, 0.5 ml/min. HA and hippuiyl-L-histidyl-L-leucine (HHL) were detected at 228 nm. For positive control, the reaction of the vial containing ACE + NaCl in buffer was immediately terminated by 1 N HCl prior to incubation. On the other hand, the vial containing ACE + NaCl in buffer was also incubated and served as negative control.
RESULTS AND DISCUSSION
Amino acid composition:
Amino acids are very important components of nutritious and functional foods and are indispensable in the human body. Aside from other common foods, mushrooms are also considered as vital sources of amino acids because of their remarkable protein content. The standard and non-standard amino acids composition varied in the different growth stages of fruiting body of V. volvacea (Table 1). All the essential and non-essential standard amino acids were present while 13 out of 21 non-standard amino acids detected in stage II, stage III and stage IV. It is interesting to note that urea was not detected in the early stage of fruiting body developmement (stage I). Generally, the concentration of amino acids increased as the fruiting bodies matured. Hence, the highest amounts of amino acids were observed at stage III and IV, which strongly suggests that the matured fruiting bodies should be consumed to obtain maximum amount of amino acids. The change in the amino acid content observed in the present study is similar to the findings of Harada et al.  who reported that free amino acids and soluble carbohydrates contents vary during fruiting body development of Hypsizygus marmoreus wherein aspartic acid, asparagine and glutamic acid peaked at stage A, while the highest amount of ornithine was found at stage B.
Standard amino acids are classified into four groups based on their characteristic taste [37, 38]. These include monosodium glutamate-like (MSG-like) amino acids (aspartic and glutamic acids), sweet taste amino acids (alanine, glycine, serine and threonine), bitter amino acids (arginine, histidine, isoleucine, leucine, methionine, phenylalanine, and valine), and tasteless amino acids (lysine and tyrosine). V. volvacea contains glutamic acid, glutamine, alanine, arginine, serine, valine, aspartic acid, threonine, lysine, tyrosine, isoleucine, cystine, asparagine, leucine, proline, glycine, histidine, phenylalanine, tryptophan and methionine, in decreasing order of abundance. Among the different standard amino acids, glutamic acid (1458.9 mg / 100 g) was the most abundant in different growth stages of fruiting body, which peaked at the final stage of development. The high glutamic acid content in the present study is in conformity with the observed amino acid content of S. commune, L. tigrinus, L. sajor-caju, C. reinakeana, C. comatus, Termitomyces globules, and Termitomyces eurrhizus, Agrocybe chaxingu, Pleurotus ostreatus, Flammulina velutipes [18, 19, 20, 25, 39]. Therefore, high glutamic acid and aspartic acid content of this mushroom dictate its pleasant and delightful taste. Accordingly, V. volvacea could be used as a natural alternative to sodium aspartate monohydrate and sodium glutamate monohydrate in enhancing umami flavour of foods.
In addition to glutamic acid, valine was also found in high concentration (335.5 mg/100 g) particularly at stage III of fruiting body development. However, this amino acid was found in small amount or even not detected in other species of mushrooms like Termitomyces sp. and Polyporus tenuiculus . The high concentration of valine in V. volvacea indicates that it can used as a substitute to animal-based products such as meat, poultry, fish, milk and daily products like cheese as which are rich sources of valine. The value obtained in the present study is higher than 11.9 to 36.6 mg / 100 g obtained by Reyes et al  in five species of edible mushrooms. Valine was also found as the most abundant essential amino acid in C. comatus, Cantharellus cibarius and Calvatia gigantean [18, 40]. In contrast, leucine was the principal component in P. ostreatus, Agaricus sp., Boletus pruinatus, Lactarius sp. [41, 42] while threonine in A. chaxingu  and phenylalanine in C. reinakeana .
Branched-chain amino acids (BCAAs) such as valine, leucine, and isoleucine, play crucial roles in the interaction of the transmembrane domains of membranous proteins with phospholipid bilayers and in shaping the structures of globular proteins . In rat, valine administration to elevate plasma valine concentrations showed biochemical and functional activities by blocking the effects of neurotransmitters of tryptophan and tyrosine which respectively stimulated lowering of blood pressure and episodic secretion of growth hormones . Moreover, BCAAs have been given in human to improve mental and physical performance and prevent the progression of central nervous system functional symptoms of neurological diseases such as hepatic encephalopathy, phenylketonuria, and bipolar disorder .
Comparing the maximum total amount of standard amino acids of V. volvacea (6070.3 mg / 100 g) with other Philippine edible and medicinal mushrooms, this paddy straw mushroom recorded the highest amount so far, followed by C. reinakeana, P. florida, and C. comatus. Evidently, these results indicate the importance of V. volvacea in addressing problem on malnutrition due to essential amino acid deficiency among children.
With regards to the non-standard amino acids, a total of 13 out of 21 non-standard amino acids were present in different stages of fruiting bodies development (Table 1). Unlike the standard amino acids, non-standard amino acids peaked at the different stages, some components increased while others declined as the fruiting body matured. However, there was an increasing pattern in the total amounts of non-standard amino acids, where stage IV registered the highest total amount (813.8 mg / 100 g). Regardless of stages, fruiting bodies of V. volvacea contained high amounts of ammonia, ornithine, urea, phosphoserine, [gamma]-aminobutyric acid (GABA). Among the different non-standard amino acids, ammonia, a naturally occurring substance present in all life forms, registered the highest concentration. This finding is in conformity with the result of Reyes et al.  who disclosed that ammonia is the predominant non standard amino acid in Philippine native and exotic species namely'. Schiophyllum commune, Lentinus tigrinus, lucidum and Pleurotus florida. In the present study, ornithine was found next to ammonia in terms of quantity. Ornithine is used as food supplement and medicine in lowering concentrations of blood ammonia and eliminating hepatic encephalopathy symptoms associated with liver cirrhosis . Nevertheless, GABA was also an important amino acid component of V. volvacea. Oral administration of GABA (0.5 mg/kg) significantly lowered the systolic blood pressure in spontaneously hypertensive rats  and consumption of food products like fermented milk with GABA showed hypotensive effects to patients with mild or moderate hypertension . Indeed, these reports may indicate promising benefits of ornithine and GABA in human well-being.
Other nutrient components:
Nutritionally, the fruiting bodies of mushrooms generally contain carbohydrate (57%), protein (25%), ash (12.5%), and fats (5.7%), on dry weight basis . In this work, the proximate nutritional components of the different growth stages of V. volvacea fruiting bodies are shown in Table 2. Obviously, stage of fruiting body was considered as an important factor on the nutrient composition as indicated by the changing amounts in every stage of development. Carbohydrate and sugars were the most abundant followed by protein. In increasing level of maturity, the concentration of carbohydrate and sugar decreased while protein increased. The maximum amount of carbohydrate at stage I (49.3 g/100 g) was within the carbohydrate contents of some wild edible mushrooms of Northern Thailand (41-65 g/100 g) studied by Sanmee et al.  and 10 popular Croatian wild edible mushroom species (42.62-66.77 g/100 g) analyzed by Beluhan and Ranogajec . In terms of soluble sugar, stage I contained the maximum amount of 43.2 g/100 g, which was higher than the total sugar contents of Calocybe gambosa, Calocybe cornucopioides, Flammulina velutipes, Macroleptiota procera, Pleurotus ostreatus, and Entoloma clypeatum . Glucose was found in higher concentration (65.51% maximum at stage III), followed by galactose,
mannose and fucose. The fair sweet taste of this mushroom could be due to the appreciable amount of sugar present in the fruiting bodies.
Protein was another notable component of this mushroom. The highest protein content (37.9 g/100 g) was noted in stage IV, while the lowest was observed in stage I (32.9 g/100 g) of fruiting body development. This increasing protein content during fruiting body development remains to be vague . However, Colak et al.  enumerated several factors that affect the protein content of mushrooms and these include part and stage of fruit body, species type, level of nitrogen in the substrate, and harvest location. Similar with protein, lipid or fat content increased in an increasing level of maturity of fruiting body. At first, the amount of fat was 3.9 g/100 g and then peaked to 4.9 g/100 g at stage IV. Values were within the range of fat contents (1.34-6.45 g/100 g) of Croatian mushrooms . This fat content could be credited to a wide variety of lipid compounds, which could be necessarily considered in future studies.
Crude fiber and ash were both peaked at stage I and stage IV of fruiting body. This maximum crude fiber content (6.1 g/100 g) was within the range of crude fiber contents (4.54-6.54 g/100 g) of edible mushrooms from South Eastern part of Nigeria . High crude fiber in food is an indicative of favourable effects on the digestion processes. The ash content (3.6 g /100 g) referred to the mineral fractions of the mushroom. Potassium (3345.21 mg/100 g at stage IV) was the most abundant mineral, followed by calcium (398 mg/100 g at stage I). Likewise, potassium (19.83-197.24 mg/100 g fresh weight) was the highest mineral component of fruiting body of selected edible mushrooms from Bangladesh  while iron (1230 mg/kg dry weight) was the most abundant in four edible mushrooms from South Western Nigeria . However, in the present study, sodium, iron, and phosphorous were found in small amounts. These findings strongly dictate that minerals of mushrooms could vary depending on species type and stage of fruiting body development.
Mushrooms are also good sources of vitamins such as riboflavin, niacin, and folates . In this work, only vitamin B complexes were detected and quantified, and among these complexes, vitamin B3 (niacin) registered the highest amount (14.56 mg/100 g peaked at stage IV) compared with vitamin B1 (thiamin) and B2 (riboflavin). Niacin plays important roles in DNA damage responses and signalling events for stress responses such as apoptosis, leading an impact on cancer risk reduction . In addition, niacin has also been used as treatment for atherosclerotic cardiovascular disease . The thiamine contents (0.37-0.44 mg/100 g) obtained in the present study were found lower than the thiamine contents (0.6-0.9 mg/100 g dry weight) of the four mushrooms assayed by Matilla et al. . On the other hand, both vitamin A and C were not found present in any stages of fruiting body of V. volvacea.
Medicinal mushrooms have been used as natural remedies for various diseases due to their bioactive metabolites. Given the wealthy bio-constituents of V. volvacea, we also investigated its functional activities such as anti-coagulant, anti-inflammatory, and anti-hypertensive activities in vitro (Figure 2) in order to establish its promising medicinal properties.
Platelet hyperaggregation is associated with atherosclerotic diseases like stroke and myocardial infarction, which is physiologically induced by several aggregating factors. The inhibitory effect of the extracts of the different stages of fruiting bodies of V. volvacea against ADP-, PAF-, and AA-Na-induced platelet aggregation was studied. Apparently, there were increasing inhibitory effects in an increasing level of maturity of fruiting body except in AA-Na, in which stage II extract showed the lowest inhibition. Extract of stage III recorded the highest inhibition of 77.3% in ADP-, 68.7% in PAF-, and 66.3% in AA-Na- induced aggregation. These platelet aggregation inhibitory values were higher than those of platelet aggregation inhibitors used in the present work as standard references and water extracts of fruiting bodies and mycelia of other mushrooms like Pholiota adipose 24004 (51.1%), Grifola frondosa 9014 (37.2%), Agaricus blazei 1174 (14.7%), Inonotus obliquus 74013 (42.2%), Fomitella fraxinea 17004 (8.3%), Sparassis crispa 150010 (26.1%), and Dictyophora indusiata with no inhibitory effect . Accordingly, V. volvacea could be a very useful candidate for medicinal or functional foods against aberrant platelet aggregation. Some novel inhibitory compounds from mushrooms have been successfully isolated and identified including 5'-deoxy-5'-methylsulphinyladenosine from Ganoderma lucidum , and a tripeptide (Trp-Gly-Cys, molecular mass of 365 Da) from I. obliquus ASI 74006 mycelia .
Inflammation is a biologic response of pro-inflammatory vascular cells to eliminate damaged cells and pathogens by releasing molecular mediators such as interleukin-1 beta (IL-1[beta]), tumor necrosis factor-alpha (TNF-[alpha]), interleukin-6 (IL-6), and interleukin-8 (IL-8) . IL-8 plays an important role on neutrophil activation as host cell defense mechanism, however, excessive IL-8 may results to several tissue damages . Moreover, IL-8 expression in cancer may contribute to tumor progression and metastasis, cancer cell growth and survival, tumor cell motion, leukocyte infiltration and modification of immune responses . So, any natural source that could restrain or inhibit IL-8 expression could be used as anti-inflammatory and anticancer agent. In the present study, expression of IL-8 decreased in the maturing fruiting body of V. volvacea extracts. Extracts of stage II and III fruiting bodies showed lower rates of IL-8 expression than the standard reference, which clearly indicates a notable effect of V. volvacea as anti-inflammatory and potential anticancer resource. Likewise, anti-inflammatory activities of some mushrooms through inhibition of other pro-inflammatory mediators have been also demonstrated. The water-soluble lyophilized oyster mushroom (Pleurotus ostreatus) concentrate (OMC) exhibited anti-inflammatory properties through the inhibition transcription factors NF-[kappa]B and AP-1  while water soluble (1->3)-, (1-4)-[beta]-D-glucans from Collybia dryophila, Lentinus edodes, and Marasmius oreades demonstrated similar activity through the inhibition of nitric oxide (NO) production in activated macrophages .
Overall, these significant fmctional activities could be attributed to the bioactive metabolites of V. volvacea, which were also highlighted in this present work. In some previous studies, essential amino acids isoleucine, leucine, and phenylalanine and vitamin B2 were linked to anti-inflammatoty activity , a nonstandard amino acid GABA to antihypertensive activity , and a semi-essential amino acid, arginine to platelet aggregation inhibition . A number of scientific investigations revealed that mushrooms have enormous potential against a wide range of human diseases due to their bioactive attributes that can be obtain through extraction . The chemical nahrre of the bioactive compounds present in mushrooms includes: polysaccharides, lipopolysaccharides, proteins, peptides, glycoproteins, nucleosides, triterpenoids, lectins, lipids and their derivatives . In addition, they are rich sources of health promoting molecules like polyphenols, flavonoids and radical scavenging properties . Because of these important components, mushrooms could act as hepatoprotective, chemopreventive, antimicrobial, antiviral, cardioprotective and immunomodulatory .
In conclusion, fruiting bodies of V. volvacea contain rich bioactive metabolites and nutritional qualities that contribute not only to its unique and delightful umami taste and aroma but most importantly to its notable functional activities. This mushroom could be considered as a novel source of potent anti-coagulant, antiinflammatory, and anti-hypertensive substances which are imperative in nutraceutical, pharmaceutical and pharmacological industry. However, further studies on the isolation and characterization of compounds responsible to these biological activities are necessary in order to establish the novel functional supremacy of this Philippine edible paddy straw mushroom.
Received 5 August 2015
Accepted 20 September 2015
Available online 30 September 2015
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(1) Fumio Eguchi, (2) Sofronio P. Kalaw, (2) Rich Milton R. Dulay, (3) Noriko Miyasawa, (4) Hiroaki Yoshimoto, (1) Tomoko Seyama and (2) Renato G. Reyes
(1) Laboratory of Forest Products Chemistry, Department of Forest Science, Tokyo University of Agriculture, Sakuragaoka, Setagayaku, Tokyo, Japan, (2) Center for Tropical Mushroom Research and Development, Department of Biological Sciences, College of Arts and Sciences, Central Luzon State University, Science City of Munoz, Nueva Ecija, Philippines, (3) Takasaki University of Health and Welfare, Japan, (4) Mush-tec Co.,Ltd.., Japan
Corresponding Author: Fumio Eguchi, Laboratory of Forest Products Chemistry, Department of Forest Science, Tokyo University of Agriculture, Sakuragaoka, Setagayaku, Tokyo, Japan. E-mail: firstname.lastname@example.org
Table 1: Comparative standard and non-standard amino acid profile of the different stages of fruiting body of V. volvacea. Standard amino acids Volvariella volvacea fruiting body (mg / 100 g) Stage 1 Stage 2 Stage 3 Alanine 549.4 466.0 85% 497.8 91% Arginine 157.2 159.7 102% 298.2 190% Asparagine 182.6 181.2 99% 213.5 117% Aspartic acid 226.8 261.0 115% 305.3 135% Cystine 163.4 152.6 93% 179.4 110% Glycine 120.0 119.7 100% 137.0 114% Glutamine 466.6 486.8 104% 676.1 145% Glutamic acid 918.9 958.3 104% 1289.1 140% Proline 143.8 146.6 102% 147.3 102% Serine 302.2 308.0 102% 340.3 113% Tyrosine 97.4 183.9 189% 228.5 235% Histidine 99.3 97.5 98% 123.7 125% Isoleucine 213.3 206.0 97% 253.7 119% Leucine 177.8 173.7 98% 165.6 93% Lysine 175.0 193.6 111% 266.7 152% Methionine 17.9 16.8 94% 16.0 89% Phenylalanine 39.5 67.7 171% 72.4 183% Threonine 240.7 233.4 97% 281.7 117% Tryptophan 46.3 59.7 129% 72.0 156% Valine 267.4 259.6 97% 335.5 125% Total 4605.5 4731.8 103% 5899.8 128% Non-standard amino acids (mg / 100 g) Phosphoserine 112.6 118.1 105% 104.3 93% Taurine N.D. N.D. N.D. Phosphoethanolamine N.D. N.D. N.D. Urea N.D. 50.2 88.3 Sarcosine N.D. N.D. N.D. [alpha]-aminoadipic acid 21.3 19.7 92% 19.3 91% Citrulline N.D. N.D. N.D. [alpha]-aminobutyric acid 3.7 3.0 81% 4.7 127% Cystathionine 24.0 27.5 115% 32.7 136% [beta]-alanine 11.4 13.0 114% 15.4 135% [beta]-aminobutyric acid N.D. N.D. N.D. [gamma]-aminobutyric acid 103.1 107.3 104% 105.1 102% (GABA) Monoethanolamine 19.8 24.9 126% 24.2 122% Ammonia 131.3 143.8 110% 188.8 144% Hydroxylysine N.D. N.D. N.D. Ornithine 50.7 67.4 133% 128.9 254% 1-methylhistidine 3.4 2.6 76% 2.9 85% 3-methylhistidine 5.6 7.2 129% 12.9 230% Anthelin N.D. N.D. N.D. Carnosine N.D. N.D. N.D. Hydroxyproline N.D. N.D. N.D. Total 486.9 584.7 120% 7275 149% Standard amino acids Volvariella volvacea fruiting body (mg / 100 g) Stage 4 Alanine 479.4 87% Arginine 356.1 227% Asparagine 196.9 108% Aspartic acid 247.5 109% Cystine 216.2 132% Glycine 137.2 114% Glutamine 626.7 134% Glutamic acid 1458.9 159% Proline 137.3 95% Serine 329.8 109% Tyrosine 266.6 274% Histidine 117.8 119% Isoleucine 241.6 113% Leucine 192.5 108% Lysine 254.3 145% Methionine 20.4 114% Phenylalanine 120.5 305% Threonine 259.4 108% Tryptophan 89.5 193% Valine 321.7 120% Total 6070.3 132% Non-standard amino acids (mg / 100 g) Phosphoserine 107.9 96% Taurine N.D. Phosphoethanolamine N.D. Urea 156.8 Sarcosine N.D. [alpha]-aminoadipic acid 20.8 98% Citrulline N.D. [alpha]-aminobutyric acid 75 203% Cystathionine 28.7 120% [beta]-alanine 13.9 122% [beta]-aminobutyric acid N.D. [gamma]-aminobutyric acid 115.4 112% (GABA) Monoethanolamine 24.7 125% Ammonia 164.7 125% Hydroxylysine N.D. Ornithine 158.7 313% 1-methylhistidine 3.3 97% 3-methylhistidine 11.4 204% Anthelin N.D. Carnosine N.D. Hydroxyproline N.D. Total 813.8 167% ND = not detected Table 2: Proximate nutritional composition of the different stages of fruiting body of V. volvacea. Nutrient content Unit Volvariella volvacea fruiting body Stage 1 Stage 2 Moisture content % 90.1 90.5 100% (fresh) Moisture content % 10.3 10.4 101% (dry) Protein g 32.9 33.1 101% Carbohydrate g 49.3 49.1 100% Sugar g 43.2 43.2 100% Crude fiber g 6.1 5.9 97% Lipid g 3.9 4.2 108% Ash g 3.6 3.2 89% Vitamin A IU ND N.D. Vitamin B1 mg 0.37 0.38 103% Vitamin B2 mg 1.33 1.29 97% Vitamin B3, niacin mg 8.96 9.64 108% Vitamin C mg N.D. N.D. Calcium mg 398.00 291.30 73% Phosphorous mg 3.42 3.31 97% Iron mg 3.61 3.33 92% Sodium mg 12.90 13.60 105% Potassium mg 2898.33 3204.34 111% Protein % 52.36 53.01 101% Neutral sugar % 46.29 45.75 99% Uronic acids % 1.35 1.24 92% Glucose % 64.69 63.61 98% Galactose % 24.33 25.67 106% Mannose % 5.35 5.47 102% Fucose % 3.98 3.58 90% Others % 1.65 1.67 101% Formic acid 3.0 3.1 103% Acetic acid 8.3 7.9 95% Lactic acid 18.1 19.3 107% Oxalic acid 124.8 132.6 106% Succinic acid 523.6 568.9 109% Fumaric acid 52.3 85.1 163% Malic acid 986.3 1253.2 127% [alpha]-ketoglutaric 58.6 77.5 132% acid Pyroglutamic acid 89.1 172.3 193% Citric acid 158.6 177.8 112% Total 2,022.7 2,497.7 123% Nutrient content Volvariella volvacea fruiting body Stage 3 Stage 4 Moisture content 90.2 100% 91.1 101% (fresh) Moisture content 10.5 102% 10.3 100% (dry) Protein 36.3 110% 38.9 118% Carbohydrate 45.2 92% 42.3 86% Sugar 39.3 91% 36.2 84% Crude fiber 5.9 97% 6.1 100% Lipid 4.6 118% 4.9 126% Ash 3.4 94% 3.6 100% Vitamin A N.D. N.D. Vitamin B1 0.44 119% 0.43 116% Vitamin B2 1.37 103% 1.36 102% Vitamin B3, niacin 12.38 138% 14.56 163% Vitamin C N.D. N.D. Calcium 324.60 82% 355.60 89% Phosphorous 3.21 94% 3.17 93% Iron 3.48 96% 3.47 96% Sodium 12.40 96% 13.10 102% Potassium 3151.70 109% 3345.21 115% Protein 56.34 108% 57.89 111% Neutral sugar 42.40 92% 40.87 88% Uronic acids 1.26 93% 1.24 92% Glucose 65.51 101% 64.83 100% Galactose 23.54 97% 24.32 100% Mannose 5.52 103% 5.38 101% Fucose 3.85 97% 3.77 95% Others 1.58 96% 1.70 103% Formic acid 3.1 103% 2.9 97% Acetic acid 7.2 87% 8.1 98% Lactic acid 20.3 112% 23.6 130% Oxalic acid 142.7 114% 154.3 124% Succinic acid 645.3 123% 724.3 138% Fumaric acid 75.3 144% 103.1 197% Malic acid 1277.8 130% 1352.4 137% [alpha]-ketoglutaric 151.3 258% 178.3 304% acid Pyroglutamic acid 184.3 207% 248.3 279% Citric acid 198.4 125% 245.7 155% Total 2,705.7 134% 3,041.0 150% ND = not detected
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|Author:||Eguchi, Fumio; Kalaw, Sofronio P.; Dulay, Rich Milton R.; Miyasawa, Noriko; Yoshimoto, Hiroaki; Seya|
|Publication:||Advances in Environmental Biology|
|Date:||Sep 1, 2015|
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