10.6092/issn.2531-7342/16457
The total amino acid compositions, total phenolic compounds, total flavonoid content, β-carotene content and hydroxyl radical scavenging activity in four wild edible mushrooms − Hericium erinaceus, Laetiporus sulphureus, Polyporus umbellatus, Sparassis nemecii have been studied in this work. Maximal indexes of total phenolic compounds in 41.28-43.82 mg of gallic acid equivalents per gram of dry weight were revealed for ethanol extracts of L. sulphureus and H. erinaceus. Maximal index of total flavonoid content in 7.91 mg of quercetin per gram of dw was revealed for ethanol extract of H. erinaceus. The content of β-carotene from ethanol extracts of studied mushrooms ranged from 2.1 to 3.1 µg β-carotene per gram of dw. The most intensive OH scavenging activity is characteristic for ethanol extract of H. erinaceus. The fraction of essential amino acids in mushrooms was 44%, 41.25%, 43.49% and 47.30% in S. nemecii, L. sulphureus, P. umbellatus and H. erinaceus, respectively. The fraction of hydrophobic amino acids ranged from 38.19% in L. sulphureus to 49.5% in H. erinaceus. Therefore, the ethanol extracts of H. erinaceus can be used as a source of compounds with high antioxidant potential. The basidiomata of S. nemecii and P. umbellatus can be recommended for use as a source of amino acids.
application/pdf
2531-7342
16457
en
Mariia Pasailiuk
https://creativecommons.org/licenses/by/4.0
Italian Journal of Mycology
wild basidiomata
Comparison of total amino acid compositions, total phenolic compounds, total flavonoid content, β-carotene content and hydroxyl radical scavenging activity in four wild edible mushrooms
Text.Serial.Journal
https://italianmycology.unibo.it/
52
Articles
2023-05-02
2023-02-14
2023-02-08
2023-05-02
112-125
Ital J Mycol
Mariia Pasailiuk
2023/05/02
10.6092/issn.2531-7342/16457
112
Hutsulshchyna National Nature Park, 84 Druzhba St., UA-78600, Kosiv, Ukraine
2531-7342
wild basidiomata
en
125
Zhuang YL, Sun LP, Zhao X, Wang JF, Hou H, Li BF (2009) Antioxidant and melanogenesis-inhibitory activities of collagen peptide from Jellyfish (Rhopilema esculentum). Journal of the Science of Food and Agriculture 89:1722–1727. https://doi.org/10.1002/jsfa.3645
Comparison of total amino acid compositions, total phenolic compounds, total flavonoid content, β-carotene content and hydroxyl radical scavenging activity in four wild edible mushrooms
https://italianmycology.unibo.it/article/download/16457/15845
52
                  IJM - Italian Journal of Mycology
                  ISSN 2531-7342 - Vol. 52 (2023): 112-125
                  Journal homepage: https://italianmycology.unibo.it/

Research article

        Comparison of total amino acid compositions, total phenolic
        compounds, total flavonoid content, β-carotene content and
          hydroxyl radical scavenging activity in four wild edible
                               mushrooms

              Halyna P. Kopylchuk1, Oksana M. Voloshchuk1, Mariia V. Pasailiuk2
1
    Yuriy Fedkovich Chernivtsi National University, 2 Kotsiubynsky St., UA-58012, Chernivtsi, Ukraine
2
    Hutsulshchyna National Nature Park, 84 Druzhba St., UA-78600, Kosiv, Ukraine

Corresponding author e-mail: mariia.pasailiuk@gmail.com


Article info
Received 14/2/2023; accepted 7/4/2023
https://doi.org/10.6092/issn.2531-7342/16457

Abstract
The total amino acid compositions, total phenolic compounds, total flavonoid content, β-carotene content
and hydroxyl radical scavenging activity in four wild edible mushrooms − Hericium erinaceus, Laetiporus
sulphureus, Polyporus umbellatus, Sparassis nemecii have been studied in this work. Maximal indexes of
total phenolic compounds in 41.28-43.82 mg of gallic acid equivalents per gram of dry weight were revealed
for ethanol extracts of L. sulphureus and H. erinaceus. Maximal index of total flavonoid content in 7.91 mg
of quercetin per gram of dw was revealed for ethanol extract of H. erinaceus. The content of β-carotene
from ethanol extracts of studied mushrooms ranged from 2.1 to 3.1 µg β-carotene per gram of dw. The most
intensive ·OH scavenging activity is characteristic for ethanol extract of H. erinaceus. The fraction of essential
amino acids in mushrooms was 44%, 41.25%, 43.49% and 47.30% in S. nemecii, L. sulphureus, P. umbellatus
and H. erinaceus, respectively. The fraction of hydrophobic amino acids ranged from 38.19% in L. sulphureus
to 49.5% in H. erinaceus. Therefore, the ethanol extracts of H. erinaceus can be used as a source of compounds
with high antioxidant potential. The basidiomata of S. nemecii and P. umbellatus can be recommended for use
as a source of amino acids.

Keywords
Hericium erinaceus, Laetiporus sulphureus, Polyporus umbellatus, Sparassis nemecii, wild basidiomata


Introduction

This study is directed at discovering natural sources of antioxidants and for the correction of various
pathological states which is typically accompanied by the intensification of free radical processes.
Fungi ‒ organisms that are the second most species-rich organism group (Dai et al., 2015) occupy
an important place in these kinds of studies. Therefore, the search for mushroom species whose
basidiomata/extracts could become a reliable source of antioxidants is an important practical
aspect of mycological science (Kozarski et al., 2015; Вoonsong et al., 2016). Mushrooms can be
also an inexhaustible source of essential amino acids, which is important in the development of
special nutritional supplements for people with various metabolic disorders and/or specific dietary
Department of Agricultural and Food Science in partnership with the Italian Mycological Society (UMI).
The journal is hosted and maintained by ABIS-AlmaDL. © 2023 The Authors. This work is licensed under CC BY 4.0
Kopylchuk et al.                                               Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



nutrition (Yuwa-Amornpitak et al., 2020). Therefore, the study of the amino acid composition of
these organisms is of great interest. Although most macromycetes have antioxidant properties and
are a source of amino acids, we give preference to species whose basidiomata are edible, (i.e. non-
poisonous for humans), currently cultivated or on the way to the development of artificial cultivation
methods, and have recorded medical and biological properties. For the reasons highlighted above, we
chose the species Laetiporus sulphureus (Bull.) Murrill, Polyporus umbellatus (Pers.) Fr., Sparassis
nemecii Pilát & Veselý (Polyporales) and Hericium erinaceus (Bull.) Pers. (Russulales).
   Laetiporus sulphureus is a fungus with positive medicinal and biological properties; it is widely
distributed across North America and Europe and occasionally a weak parasite. This species is
edible when young; its basidiomata grow striking golden-yellow shelf-like structures on tree trunks
and branches. This mushroom produces the L. sulphureus lectin, which exhibits haemolytic and
haemagglutination activities (Wang et al., 2018). Commercial cultivation of L. sulphureus occurs but
at a much smaller scale than the oyster mushroom and Agaricus bisporus (J.E. Lange). The resulting
mature basidiomata can reach a weight of 200‒300 g (Pleszczyńska et al., 2012).
   The chemical composition of the fungus was studied partly. Kovács and Vetter (2015) reported
macrocomponents (crude protein, -fibre, -fat, -ash contents, calculated carbohydrate concentration,
organic ingredients and the energy level), digestible protein, non-protein nitrogen and free amino
acid contents (not amino acid list) of basidiomata of L. sulphureus. Mineral components, different
protein fractions, soluble oligo- and polysaccharides of L. sulphureus were studied too. Sułkowska-
Ziaja et al. (2018) reported proteins and fats content, total carbohydrates, malic-, citric- and ascor-bic
acid of the basidiomata. The selected compounds isolated from L. sulphureus revealed anticoagulant,
hypoglycemics, hypolipemic, anticancer, cytotoxic, anti-inflammatory, acetylcholinesterase
inhibiting, antioxidants, antifungal and reverse transcriptase of HIV-virus, inhibitors activities.
Therefore, the study of amino acid content and antioxidant activity of ethanol extracts of basidiomata
of L. sulphureus will complement data about the biological value of the fungus.
   Polyporus umbellatus is an edible and medicinal mushroom. The young basidiomata growing on
the ground are edible, while its underground part has medicinal properties (Jong and Birmingham,
1990; Zhao and Zhang, 1992). This fungus is widely distributed in Europe, North America, and
East Asia (Bandara et al., 2015). This mushroom is considering promising for cultivation due to its
medicinal, and nutritional value. It is popular in China’s fungal market and is called “Zhu Ling”.
Numerous publications are devoted to the development of methods of cultivation of P. umbellatus,
in particular sclerotia and/or basidiomata of the fungus (Huang and Liu, 2007; Xing et al., 2013;
Pasailiuk, 2020).
   The medical and biological properties of P. umbellatus are well studied. Food and dietary supplements
have been developed based on the mycelium and sclerotia of this fungus. Medical attributes of this
fungus include enhanced liver function (Complexe Hepato Bio), relief of inflammatory conditions,
supporting immune function (Polyporus MRL preparation), supporting white blood cell production
and providing antioxidant protection (Mushroom 6 Immune Support Complex), strengthing the
immune system (Mycophyto ® Complex), enhancing the body’s immune response to an antigen
(P. umbellatus Extract); it is also suitable for diabetics and people who suffer from celiac disease
(P. umbellatus HdT), those who are diuretic, and enhances anticancer activity (Zhu Ling Hot-water
extract).
   The basidiomata of P. umbellatus are a source for biological active supplements such as “P.
umbellatus 180 capsules” which have antitumor effects, immune system enhancement, hair
growth, antioxidant and free-radical scavenging activity, diuretic effects, and antiviral effects
(Bandara et al., 2015). Basidiomata of fungus are rich in important bioactive substances such as
fatty acids, 2-hydroxytetracosanoic acid (Yosioka and Yamamoto, 1964), (1→3)-β-D-glucan (Lee

                                                   113
Kopylchuk et al.                                              Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



and Park, 2001), polysaccharides (Dai et al., 2012). Lu et al. (1985) isolated four components, viz.
ergosta-5,7,22-trien-3-ol (ergosterol), ergosta-7,22-dien-3-one, ergosta-7,22-dien-3-ol, and 5α,8α-
epidioxyergosta-6,22-dien-3-ol, from the basidioma of P. umbellatus. Ohsawa et al. (1992) identified
seven polyporusterones from the basidiomata of P. umbellatus and named them as A, B, C, D, E, F
and G. However, the amino acid content of P. umbellatus basidiomata has not been studied. Since
protein content has a direct effect on free radical scavenging activity, we decided to study amino acid
composition and antioxidant activity of wild P. umbellatus basidiomata.
   Sparassis nemecii is an edible mushroom. It was included to the Red Lists of some European
and Asian countries under an Endangered status. Examples of the studied fungus were found on the
territory Hutsulshchyna National Nature Park (Hutsulshchyna NNP). It is the only place of fungus
growth in Hutsulshchyna NNP and in Ukraine. Fungi were identified by Heluta et al. (2016). Pure
cultures of this fungus are stored in the IBK Mushroom Culture Collection (Bisko et al., 2016, https://
ccinfo.wdcm.org/details?regnum=1152) and in the mycological laboratory of Hutsulshchyna NNP.
Cultivation of the fungus was done by Pasailiuk (2019). Suitable substrates for growing mycelium
and small size basidiomata of S. nemecii were established. The mechanical properties of the substrate
were also important factors for the growth of S. nemecii mycelium.
   Sparassis spp., contribute a significant economic and industrial role as a source of health supplement
therapeutics, pharmaceutics, and biotechnological products. There is a close relationship between
Sparassis species and L. sulphureus, which also produces brown rot, and was suggested by sequence
data from rDNA (including the ITS region) and the partial gene coding RNA polymerase sub-unit II
(RPBII) (Wang et al., 2004). However, the chemical composition of S. nemecii, its medicinal value and
commercial value were not studied. Consequently, it is important to study amino acid compositions
and antioxidant activity of ethanol extracts of wild basidiomata of fungus, which we do in this article.
   Hericium erinaceus is an edible and medicinal mushroom that belongs to the Russulalles order. This
mushroom is very popular in Eastern medicine and the methods of its cultivation have been studied
(Figlas et al., 2007; Hassan, 2007; Julian et al., 2018). For this reason, some of its properties and
bioactive substances have already been studied. From the basidiomata of the mushroom, H. erinaceus,
biological active components were isolated as meroterpenoid, named hericenone K, astradoric acid
C, 3,4-dihydro-5-methoxy-2-methyl-2-(4’-methyl-2’-oxo-3’-pentenyl)-9(7H)-oxo-2H-furo[3,4-h]
benzopyran, ergosterol peroxide, 3β,5α,9α-trihydroxy-ergosta-7,22-dien-6-one, cerevisterol,
inoterpene A, ursolic acid, betulin, hemisceramide, and oleanolic acid (Zhang et al., 2015). The ability
of H. erinaceus mycelia to inhibit bacterial growth was reported in Julian et al. (2018). This fungus
(both the mycelia and basidiomata) has antidiabetic, antihyperlipodemic, hepatoprotective, anti-
inflammatory, anticarcinogenic, antioxidative, antihypertensive, cardioprotective, antisenescence,
antifatigue, antibiotic, immunostimulating, nephroprotective, and neuroprotective properties. It
can improve anxiety, cognitive function, and lower depression levels (Friedman, 2015). In Eastern
Asia, H. erinaceus is used traditionally to improve memory (Zhang et al., 2017). Considering the
high medical and edible value of the mushroom, information about its amino acid composition and
antioxidant activity of alcohol extracts of the wild H. erinaceus basidiomata of is an important aspect
for understanding the nutritional value of the mushroom.
   The purpose of the work is to study the amino acid composition and antioxidant activity the ethanol
extracts of wild basidiomata of S. nemecii, L. sulphureus, P. umbellatus and H. erinaceus mushrooms.




                                                  114
Kopylchuk et al.                                             Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



Materials and Methods

Source of the basidiomata
The fungi studied, S. nemecii, L. sulphureus, P. umbellatus and H. erinaceus, were collected on the
territory of Hutsulshchyna NNP. Fruiting body samples of the fungi were included in the collection
of dry and wet prepaprations at Hutsulshchyna NNP. Sparassis nemecii was introduced to a pure
culture and transferred to the IBK mushroom culture collection at the M.G. Kholodny Institute of
Botany, National Academy of Sciences (NAS) of Ukraine, Kyiv, Ukraine (Bisko et al., 2016; https://
ccinfo.wdcm.org/details?regnum=1152). Mushroom samples were dried, minced, and stored in glass
containers prior to the extraction process.

Preparation of mushroom extracts
Extraction was performed according to the method outlined in Boonsong et al. (2016) with some
modifications. For 70% (v/v) ethanol extraction, each powdered sample (5 g) was mixed with 50 mL
of 70% (v/v) ethanol and shaken at 150 rpm at room temperature for 24 h; it was then centrifuged
at 12,000 rpm for 15 min. The supernatant was filtered with Whatman filter paper and the filtrate
was collected. The residue was re-extracted under the same conditions. The obtained extract was
concentrated under vacuum at 40 ºC using the rotary evaporator. The obtained sample was stored at
4 ºC.

Determination of total phenolic compounds in mushroom extracts
Phenolic compounds in the sample extracts were estimated by using Folin-Ciocalteu assay, based on
procedures described by Gan et al. (2013). One milliliter of sample from ethanol extract was mixed
with 1 mL of Folin-Ciocalteu’s phenol reagent (1:9; Folin-Ciocalteu’s reagent: distilled water). After
5 min, 1 mL of 13% sodium carbonate solution was added to the mixture and adjusted to 10 mL with
distilled water. The reaction was kept in the dark for 90 min and its absorbance was read at 725 nm. A
calibration curve was constructed with different concentrations of gallic acid as standard. The results
were expressed as mg of gallic acid equivalents per gram of dry weight (mg GAE g-1 dw).

Determination of total flavonoid content
The content of total flavonoid of each ethanolic mushroom extract was determined using an aluminum
chloride colorimetric assay, as described previously (Kumari et al., 2011). One mL of extract was
diluted using 70% aqueous ethanol. 100 µL 10% aluminum nitrate and 100 µL, 1 M aqueous potassium
acetate were mixed into the solution and incubated at room temperature for 40 min. The absorbance
was recorded at 510 nm using a spectrophotometer (Agilent Technologies, USA). Quercetin was used
as a standard to calculate the concentration of total flavonoids. Data were calculated as milligrams of
quercetin equivalents per gram of dry weight (mg QE g-1 dw).

Determination of β-carotene
β-Carotene was determined from the dried ethanol extract according to Kumari et al. (2011). 100
mg of extract was mixed with 10 mL of acetone-hexane mixture (4:6) for 1 min and filtered. The
absorbance was recorded at three different wavelengths (453, 505 and 663 nm). The β-carotene
content was calculated by:
   β-carotene (µg g-1 dw) = 0.216 × A66 3 − 0.304 × A505 + 0.452 × A453

Hydroxyl radical scavenging activity
Hydroxyl radical scavenging activity of the extractives was determined by the method of Halliwell and

                                                 115
Kopylchuk et al.                                            Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



Gutteridge (1989) with minor modifications (Rahman et al., 2015). Hydroxyl radical was generated
by the Fe3+-ascorbate-EDTA-H2O2 system (Fenton reaction). The assay is based on the quantification
of the 2-deoxy-D-ribose degradation product, which forms a pink chromogen upon heating with TBA
at low pH.
    The reaction mixture contained deoxyribose (2.8 mM), KH2PO4-NaOH buffer, pH 7.4 (0.05 M),
FeCl3 (0.1 mM), EDTA (0.1 mM), H2O2 and mushroom extracts in a final volume of 2 mL. The
mixture was incubated at 37°C for 30 min followed by the addition of 2 mL of trichloroacetic acid
(2.8% w/v) and thiobarbituric acid. Thereafter, it was kept for 15 min in a boiling water bath, and
cooled. The absorbance of solution was measured at 532 nm with a spectrophotometer. The hydroxyl
radical scavenging capacity was evaluated with the inhibition of percentage of 2-deoxy-D-ribose
oxidation on hydroxyl radicals.
    The percentage of hydroxyl radical scavenging activity was calculated according to the following
formula:

   % hydroxyl radical scavenging activity = [A0 − (A1 − A2)] × 100 / A0

   Where:
   A0 is the absorbance of the control without a sample,
   A1 is the absorbance after adding the sample and 2-deoxy-D-ribose,
   A2 is the absorbance of the sample without 2-deoxy-D-ribose.

AAs analysis
For the analysis of hydrolyzed amino acids (AAs), the samples were hydrolyzed in 6 N hydrochloric
acid in vacuum-sealed tubes at 106 ºC for 24 h. The AAs content of the various hydrolysates was
determined using an AAs automatic analyzer of amino acids T-339 (Microtechnology, Czech
Republic, on the basis of Palladin Institute of Biochemistry of the NAS of Ukraine) with postcolumn
derivatization using ninhydrin. The concentrations of the amino acids (in g/100 g proteins) were
calculated from external standards for the different amino acids.

Data analysis
The results were processed using Statistica 8.0 (StatSoft Inc., Tulsa, Oklahoma, USA). All experiments
were conducted using three biological replicates. All values are mean ± SD (n = 3).

Results

Determination of total phenolic compounds
The obtained results showed significant differences in the content of total phenolic compounds in the
ethanol extracts of the studied mushrooms. The maximum indicators of total phenolic compounds
range from 41.28 to 43.82 mg GAE g-1 dw and were registered for L. sulphureus and H. erinaceus
samples. A significantly lower content of total phenolic compounds was noted in samples of P.
umbellatus and S. nemecii. The minimum values of total phenolic compounds recorded in S. nemecii
were 6.67 mg GAE g-1 dw, which is about 6.2-6.5 times less compared to the established maximum
values in the samples of H. erinaceus and L. sulphureus. The content of total phenolic compounds in
mushroom extracts is shown in Figure 1.




                                                 116
Kopylchuk et al.                                                                 Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457




                                                             A
                                  50
                                                                                         A
                                  40
                mg GAE/g dw




                                  30



                                  20            C                           B

                                  10



                                  0
                                         S. nemecii    L. sulphureus P. umbellaus    H. erinaceus
          Fig. 1 ‒ Total phenolic compounds in ethanol extracts of Sparassis nemecii, Laetiporus sulphureus,
          Polyporus umbellatus and Hericium erinaceus basidiomata. All values are Mean ± SD (n = 3). Values
          bearing different letters in the same column are significant at P<0.05.

Determination of total flavonoid content
Total flavonoid content in the studied mushrooms is shown in Figure 2. Ethanol extracts of P.
umbellatus and S. nemecii revealed that samples of these fungus had a lower level of total flavonoid
content (4.3-4.6 mg QE g-1 dw.) than other studied mushrooms. The maximal total flavonoid content
was registered in ethanol extract of H. erinaceus– 7.91 mg QE g-1 dw.


                       10
                                                                                             A

                                                             B
   mg QE/g dw




                                            C                               C
                              5




                              0
                                       S. nemecii     L. sulphureus   P. umbellaus    H. erinaceus
 Fig. 2. − Total flavonoid content in ethanol extracts of Sparassis nemecii, Laetiporus sulphureus, Polyporus umbellatus
 and Hericium erinaceus basidiomata.


                                                                      117
Kopylchuk et al.                                                            Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



Determination of β-carotene
The content of β-carotene in extracts of the basidiomata of the studied mushrooms did not differ
significantly and ranged from 2.1 to 3.1 µg g-1 dw (Fig. 3).

                         4
                                                                                     A
                                                       A
                                     B
                                                                       B
        µg/g dw




                         2




                         0
                                  S. nemecii       L. sulphureus P. umbellaus     H. erinaceus
     Fig. 3. − Content of β-carotene in ethanol extracts of Sparassis nemecii, Laetiporus sulphureus, Polyporus
     umbellatus and Hericium erinaceus basidiomata.

Hydroxyl radical scavenging activity
Investigation of the hydroxyl radicals scavenging activity of ethanol extracts of mushroom basidiomata
showed that the most intense absorption of ·OH radicals is caused by ethanol extracts of H. erinaceus
which inhibits ·OH radicals by 67.7%. Indicators of hydroxyl radical scavenging activity of mushroom
extracts are shown in Figure 4.

                         100

                                                                                         A A
                             75
                                         B                             B
                                               B                           B
                                                         C
          % inhibition




                             50



                             25



                             0
                                    S. nemecii       L. sulphureus P. umbellaus    H. erinaceus
         Fig. 4. − Hydroxyl radical scavenging activity of ethanol extracts of Sparassis nemecii, Laetiporus
         sulphureus, Polyporus umbellatus and Hericium erinaceus basidiomata.


                                                                 118
Kopylchuk et al.                                                       Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



AAs analysis
Amino acid content was analyzed in the studied mushrooms. Table 1 shows the amino acid profile of
the four mushroom species in g/100g of protein. For S. nemecii and P. umbellatus, we recorded the
highest content of five amino acids, in particular lysine, aspartic acid, glutamic acids, leucine, and
isoleucine. Our research shows that H. erinaceus has the lowest amount of all amino acids except for
three (aspartic acid, glutamic acids, leucine). In L. sulphureus, the content of amino acids is lower
compared to S. nemecii and P. umbellatus, but significantly higher than in H. erinaceus.

Table 1 - Amino acid composition of Sparassis nemecii, Laetiporus sulphureus, Polyporus umbellatus and Hericium
erinaceus mushrooms.

AAs                                      Mushroom
                                         S. nemecii            L. sulphureus       P. umbellatus      H. erinaceus
Lysine*                                  1.74±0.065a           1.10±0.028b         1.47±0.107a        0.22±0.011c
Histidine*                               0.50±0.046a           0.27±0.033b         0.42±0.021a        0.06±0.002c
Arginine                                 1.29±0.061a           0.75±0.054b         1.23±0.037a        0.14±0.019c
Aspartic acid                            2.37±0.058a           1.53±0.102b         2.01±0.116a        0.37±0.044c
Threonine*                               1.18±0.041a           0.67±0.037b         0.97±0.124a        0.14±0.023c
Serine                                   1.07±0.085a           0.68±0.058c         0.87±0.112b        0.15±0.037d
Glutamic acid                            3.15±0.073a           2.02±0.016b         2.98±0.212a        0.53±0.071c
Proline                                  0.99±0.066a           0.54±0.048b         0.84±0.131a        0.27±0.012c
Glycine                                  1.12±0.078a           0.71±0.069c         0.97±0.086b        0.18±0.017d
Alanine                                  1.54±0.083a           1.09±0.099b         1.34±0.089a        0.24±0.031c
Cystine                                  0.47±0.049a           0.29±0.032b         0.23±0.041c        0.07±0.009d
Valine*                                  1.15±0.120a           0.81±0.062b         1.06±0.105a        0.17±0.041c
Methionine*                              0.22±0.014a           0.15±0.011b         0.10±0.009c        0.09±0.005c
Isoleucine*                              2.12±0.113a           1.08±0.071b         1.93±0.141a        0.77±0.066c
Leucine*                                 1.90±0.077a           1.15±0.027c         1.64±0.096b        0.31±0.014d
Tyrosine                                 0.51±0.047a           0.14±0.011c         0.35±0.073b        0.09±0.011d
Phenylalanine*                           1.06±0.056a           0.55±0.048b         0.90±0.094a        0.16±0.012c
Gamma-Aminobutyric Acid (GABA)           0.04±0.006d           0.49±0.052a         0.23±0.019b        0.09±0.011c
TAA, total amino acid                    22.41                 14.01               19.52              4.06
ЕАА/ТАА                                  44.00 %               41.25 %             43,49 %            47.30 %
НАА/ТАА                                  40.00 %               38.19 %             40.00 %            49.50 %

Notes – All values are means ± SD of three determinations (in g/100 g protein). Means with the same letter across a row
are not significantly different at 0.05 probability level. *Essential amino acids (ЕАА); ТАА, total amino acid; HAA,
hydrophobic amino acids.

    Based on the results of this study, the four mushroom species can be ranked in order of decreasing
amino acid content: S. nemecii > P. umbellatus > L. sulphureus > H. erinaceus. In addition, studies
have shown that content of amino acids such as aspartic acid and glutamic acids is in maximum
quantities, while the lowest content is methionine, tyrosine, histidine and cystine. The glutamic acid
content varied from 3.15 g/100 g in S. nemecii to 0.53 g/100 g in H. erinaceus, while the methionine
content varied from 0.22 g/100 g in S. nemecii to 0.09 g/100 g in H. erinaceus. Laetiporus sulphureus
is enriched in GABA (0.49 g/100 g) compared to other studied mushroom species.


                                                         119
Kopylchuk et al.                                              Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



   The studied species of mushrooms contain all essential amino acids. The fraction of essential
amino acids in mushrooms was 44.00%, 41.25%, 43.49%, and 47.30% in S. nemecii, L. sulphureus,
P. umbellatus, and H. erinaceus, respectively. At the same time, leucine and isoleucine predominate
among essential amino acids in all studied mushrooms. The proportion of hydrophobic amino acids
(Ala, Pro, Val, Met, Phe, Ile, and Leu) ranged from 38.19% in L. sulphureus to 49.50% in H. erinaceus.

Discussion

Currently, the issue of finding natural sources of antioxidants that can be used to correct various
pathological conditions accompanied by the intensification of free radical processes remains
relevant. Therefore, the demand for discovering novel compounds with good antioxidant activity is
understandable (Zhou et al., 2010). In the context of this problem, mushrooms are increasingly being
considered as potential producers of antioxidants. Mushrooms are also important as a nutraceutical and
dietary support (Turfan et al., 2020). In this study, we evaluated/analyzed the amino acid composition,
total phenolic compounds, total flavonoid content, β-carotene and antioxidant activity of ethanol
extracts of four mushroom species S. nemecii, L. sulphureus, P. umbellatus and H. erinaceus. Ethanol
as an extractant was chosen for the reason established by Jiang et al. (2016); anhydrous ethanol
appears to be the most efficient extraction solvent that can be used to extract the effective antioxidant
compounds in the pharmaceutical and food industries.
    A number of studies have demonstrated the close association between the prevention of reactive
oxygen species-associated diseases (ROS-associated diseases) and intake of food rich in antioxidants,
including mushrooms (Abdullah et al., 2012). A vast body of evidence indicates that wild edible
mushrooms contain many biologically active compounds disclosing antioxidant and other properties
(Sun et al., 2017). Among the components of mushrooms that exhibit high antioxidant activity, it is
worth noting carotenoids, in particular β-carotenes, polyphenols, including flavonoids (Podkowa et
al., 2021).
    Carotenoids protect fungi from oxidative stress and non-ionizing irradiation such as UV light
(Lin and Xu, 2020). Therefore, they act as photoprotectors against the lethal combination of light
and as antioxidants (Ribeiro et al., 2011). Carotenoids are known as singlet oxygen quenchers and
lipid peroxidation chain breakers (Robaszkiewicz et al., 2010). Additionally, carotenoids are among
the bioactive products with significant medical value (Mata-Gómez et al., 2014). In 1976, Valadon
reported the carotenoids are additional taxonomic characters in fungi. In a number of cases, they are
very good taxonomic markers. According to Zhang et al. (2018), the content of the carotenoids is one
of the indicators of the quality of commercial basidiomata, for example Cordyceps militaris (L.) Fr.
Information on carotenoids in mushrooms is fragmentary. Robaszkiewicz et al. (2010) reported the
content of β-carotene differed considerably between the thirteen analyzed edible mushroom species
from 0.233 (Tuber mesentericum Vittad.) to 18.649 μg g-1 (Tricholoma equestre (L.) P. Kumm.) of
dried body. For dried vegetables, for examples carrot and pumpkin β-carotene reveals 26.57 and
773.66 µg g-1 dw (Piyarach et al., 2020). The content of β-carotene in extracts of the fungi in our study
showed that basidiomata of the mushrooms ranged from 2.1 to 3.1 µg g-1 dw. These indexes are higher
than, for example, indexes of β-carotene methanolic extract of Boletus edulis Bull. estimated by the
same method (Robaszkiewicz et al., 2010).
    The high content of phenolic compounds in ethanol extracts of L. sulphureus and H. erinaceus,
along with the high content of flavonoids and β-carotene, can be considered as an indicator of their
potential antioxidant activity. This probably explains the high hydroxyl inhibitory activity of H.
erinaceus mushroom ethanol extracts. The hydroxyl radical is the most reactive among ROS due to
its high oxidizing activity. Therefore, the neutralization of hydroxyl radicals is considered as the most

                                                  120
Kopylchuk et al.                                              Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



effective way to protect against oxidative damage to cell biomolecules. Antioxidants of fungi can
exert their effect in two ways: the first consists in the direct absorption of hydroxyl radicals, while
the second is in the suppression of their generation (Zhang et al., 2011). The antioxidant activity of
mushrooms is directly proportional to the amount of bioactive compounds (Elbatrawy et al., 2015).
Antioxidant activity is mainly associated with the content of phenol, but the carotenoids, vitamins,
and polysaccharides present in the extracts enhance the antioxidant effects (Palacios et al., 2011). The
antioxidant activity of phenolic compounds depends on the number of functional groups and location.
It is believed that an important factor affecting the antioxidant activity of phenolic compounds is the
number of hydroxyl groups, because their ability to chelate metal ions depends on this (Hussain et
al., 2016). The main mechanism of antioxidant action of phenolic compounds consists either in direct
inhibition of free radicals or in strengthening of endogenous antioxidant activity. In addition, some
phenolic compounds stimulate the synthesis of endogenous antioxidants through the activation of the
Nrf/ARE pathway (Scalbert et al., 2005). The reaction of cells to these compounds is implemented
through direct interaction with enzymes or receptors involved in signal transmission, which leads to
modification of the redox state of the cell and can cause a number of redox-dependent reactions. As
antioxidants, polyphenols increase the probability of cell survival, but under the conditions of their
excessive intake, they can show pro-oxidant properties (Kozarski et al., 2011).
    The mechanisms of antioxidant action of flavonoids include direct scavenging of free radicals,
chelation of metal ions involved in free radical formation, inhibition of enzymes such as xanthooxidases
and lipoxygenases involved in free radical production, and regeneration of membrane-bound
antioxidants. such as α-tocopherol (Kozarski et al., 2011). It is asserted that the main mechanism of
action of flavonoids on the absorption of radicals is the donation of a hydrogen atom (Amic et al.,
2007). The ability of flavonoids to inhibit ROS depends on the location and number of hydroxyl groups
in the molecule. The double bond and carbonyl function in the heterocycle or polymerization of the
nuclear structure increases activity by affording a more stable flavonoid radical through conjugation
and electron delocalization (Heim et al., 2002). Pan et al. (2016) show that flavonoids are powerful
scavengers of hydroxyl and superoxide radicals and actively chelate transition metals. However,
despite the high content of phenolic compounds, flavonoids and β-carotenes in L. sulphureus, we
found low hydroxyl inhibitory activity for alcohol extracts of this fungus. Probably, the antioxidant
compounds of this mushroom are involved in the process of neutralization of other ROS.
    The high antioxidant capacity can be confirmed by the results of our amino acid analysis. Some
studies have shown that hydrophobic amino acids (HAA) play an important role in the manifestation
of antioxidant activity (Sun et al., 2017). A high content of HAA could enhance their antioxidant
activity (Zhuang et al., 2009). Thus, it is possible to expect a high antioxidant activity of the studied
mushrooms, especially mushrooms of the H. erinaceus species, which have the highest content of
hydrophobic amino acids. The amino acid composition of mushrooms is a reliable indicator of the
nutritional value of food (Sun et al., 2017). At the same time, the results of the amino acid analysis
obtained by us do not provide grounds for asserting the high nutritional value of the H. erinaceus
mushroom, since the samples of the basidiomata of this mushroom are characterized by a low total
content of amino acids compared to other studied mushroom samples. With regard to the species
S. nemecii and P. umbellatus, which are characterized by a low content of phenolic compounds,
flavonoids and β-carotenes and a lower hydroxyl inhibitory activity compared to H. erinaceus, but
we can state their high nutritional value based on the amino acid analysis based on our study. These
mushrooms are characterized by a high total content of amino acids with a significant proportion of
all essential amino acids.




                                                  121
Kopylchuk et al.                                              Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



Conclusion

Therefore, H. erinaceus, which ethanol extracts showed the highest content of antioxidant compounds
and the highest hydroxyl inhibitory activity, can be used as a source of compounds with high
antioxidant potential. Its antioxidant compounds may be prospective protective agents that will help
prevent and treat oxidative stress-associated diseases. Sparassis nemecii and P. umbellatus can be
recommended for use as a potential source of amino acids, including essential ones, as well as for the
development of nutritional supplements for people with various metabolic disorders and/or specific
dietary nutrition.

Acknowledgements

The authors express their sincere gratitude to Nina M. Fontana (Department of Native American
Studies, University of California, Davis, California 95616) for editorial assistance with the manuscript.

References

Abdullah N, Ismail SM, Aminudin N, Shuib AS, Lau BF (2012) Evaluation of selected culinary-
      medicinal mushrooms for antioxidant and ACE inhibitory activities. Evidence-Based
      Complementary and Alternative Medicine 2012:464238. https://doi.org/10.1155/2012/464238
Amic D, Davidovic-Amic D, Beslo D, Rastija V, Lucic B, Trinajstic N (2007) SAR and QSAR of
      the antioxidant activity of flavonoids. Current Medicinal Chemistry 14:827–45. https://doi.
      org/10.2174/092986707780090954
Bandara AR, Rapior S, Bhat DJ, Kakumyan P, Sunita C, Xu J, Hyde KD (2015) Polyporus umbellatus,
      an edible-medicinal cultivated mushroom with multiple developed health-care products as
      food, medicine and cosmetics: a review. Cryptogamie, Mycologie 36(1): 3‒43. https://doi.
      org/10.7872/crym.v36.iss1.2015.3
Bisko NA, Lomberg ML, Mytropolska NY, Mykchaylova OB (2016) The IBK Mushroom Culture
      Collection. Alter press, Kyiv.
Вoonsong S, Wanwimol K, Pongtep W (2016) Antioxidant cctivities of extracts from five edible
      mushrooms using different extractants. Agriculture and Natural Resources 50:89−97. https://
      doi.org/10.1016/j.anres.2015.07.002
Dai YC, Cui BK, Si J, He SH, Hyde KD, Yuan HS, Liu XY, Zhou LW (2015) Dynamics of the
      worldwide number of fungi with emphasis on fungal diversity in China. Mycology Progress
      14(62):1–9. https://doi.org/10.1007/s11557-015-1084-5
Dai H, Han XQ, Gong FY, Dong H, Tu PF, Gao XM (2012) Structure elucidation and immunological
      function analysis of a novel beta-glucan from the fruit bodies of Polyporus umbellatus (Pers.)
      Fries. Glycobiology 22:1673−1683. https://doi.org/10.1093/glycob/cws099
Elbatrawy EN, Ghonimy EA, Alassar MM, Wu FS (2015) Medicinal mushroom extracts possess
      differential antioxidant activity and cytotoxicity to cancer cells. International Journal of
      Medicinal Mushrooms 17(5):471−479. https://doi.org/10.1615/intjmedmushrooms.v17.i5.70
Figlas D, González M, Ramiro CN (2007) Cultivation of culinary-medicinal Lion’s Mane mushroom
      Hericium erinaceus (Bull.: Fr.) Pers. (Aphyllophoromycetideae) on substrate containing
      sunflower seed hulls. International Journal of Medicinal Mushrooms 9:67−73. https://doi.
      org/10.1615/IntJMedMushr.v9.i1.80




                                                  122
Kopylchuk et al.                                            Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



Friedman M (2015) Chemistry, nutrition, and health-promoting properties of Hericium erinaceus
      (Lion’s Mane) mushroom fruiting bodies and mycelia and their bioactive compounds. Journal of
      Agricultural and Food Chemistry 63(32):7108−7123. https://doi.org/10.1021/acs.jafc.5b02914
Gan CH, Nurul AB, Asmah R (2013) Antioxidant analysis of different types of edible mushrooms
      (Agaricus bisporous and Agaricus brasiliensis). International Food Research Journal
      20(3):1095−1102.       http://www.ifrj.upm.edu.my/20%20(03)%202013/8%20IFRJ%2020%20
      (03)%202013%20Asmah%20(306).pdf
Halliwell B, Gutteridge JMC (1989) Free radicals in biology and medicine. Clarendon Press 3:617–
      683.
Hassan F (2007) Cultivation of the monkey head mushroom (Hericium erinaceus) in Egypt. Journal
      of Applied Sciences Research 3(10):1229−1233.
Heim KE, Tagliaferro AR, Bobilya DJ (2002) Flavonoid antioxidants: chemistry, metabolism and
      structure-activity relationships. The Journal of Nutritional Biochemistry. 13(10):572-584.
      https://doi.org/10.1016/S0955-2863(02)00208-5
Heluta VP, Fokshei SI, Derzhypilskyi LM (2016) The first finds in Ukraine of the rare mushroom
      Sparassis nemecii (Sparassidaceae). In: Rare plants and fungi of Ukraine and adjacent areas:
      implementing conservation strategies (Didukh, ed), 16‒20 May 2016, Palivoda, Kyiv, Ukraine,
      pp 182−183. [in Ukraine]
Huang HC, Liu YC (2007) Acceleration of mycelia growth by adding Grifola umbellata broth
      concentrate in solid-state and submerged cultures. Biochemical Engineering Journal 37:139−143.
      https://doi.org/10.1016/j.bej.2007.04.003
Hussain T, Tan B, Yin Y, Blachier F, Tossou M, Rahu N (2016) Oxidative stress and inflammation:
      What polyphenols can do for us? Oxidative Medicine and Cellular Longevity 2016:7432797.
      https://doi.org/10.1155/2016/7432797
Jiang S, Wang Y, Zhang X (2016) Comparative studies on extracts from Hericium erinaceus by
      different polarity reagents to gain higher antioxidant activities. Experimental and Therapeutic
      Medicine 12(1):513−517. https://doi.org/10.3892/etm.2016.3279
Jong S, Birmingham J (1990) The medicinal value of the mushroom Grifola. World Journal of
      Microbiology and Biotechnology 6:227−235. https://doi.org/10.1007/BF01201290
Julian A, Wright C, Reyes R (2018) Prelude to successful cultivation of Hericium in the Philippines:
      understanding its mycelial growth response on different culture media and its antibacterial
      activity. International Journal of Pharmaceutical Research and Allied Sciences 7:1−7. https://
      doi.org/10.4489/MYCO.2008.36.2.088
Kovács D, Vetter J (2015) Chemical composition of the mushroom Laetiporus sulphureus (Bull.)
      Murill. Acta Alimentaria 44:104−110. https://doi.org/10.1556/AAlim.44.2015.1.10
Kozarski M, Klaus A, Jakovljevic D, Todorovic N, Vunduk J, Petrović P, Niksic M, Vrvic MM,
      van Griensven L (2015) Antioxidants of edible mushrooms. Molecules 20(10):19489−19525.
      https://doi.org/10.3390/molecules201019489
Kozarski M, Klaus A, Niksic M, Jakovljevic D, Helsper JPFG, van Griensven LJLD (2011)
      Antioxidative and immunomodulating activities of polysaccharide extracts of the medicinal
      mushrooms Agaricus bisporus, Agaricus brasiliensis, Ganoderma lucidum and Phellinus
      linteus. Food Chemistry 129:1667–1675. https://doi.org/10.1016/j.foodchem.2011.06.029
Kumari D, Reddy MS, Upadhyay RC (2011) Antioxidant activity of three species of wild mushroom
      genus Cantharellus collected from North-Western Himalaya, India. International Journal of
      Agriculture and Biology 13:415–418.
Lee YK, Park JH (2001) December 08. Extraction of pharmacologically active polysaccharide (beta-
      glucan) fraction from Polyporus umbellatus. South Korea patent application KR2001108709A.
      2001.12.08.
                                                123
Kopylchuk et al.                                            Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



Lin L, Xu J. (2020) Fungal pigments and their roles associated with human health. Journal of Fungi
      6(4):280. https://doi.org/10.3390/jof6040280
Lu W, Adachi I, Kano K, Yasuta A, Toriizuka K, Ueno M, Horikoshi I (1985) Platelet aggregation
      potentiators from Cho-Rei. Chemical and Pharmaceutical Bulletin 33:5083−5087. https://doi.
      org/10.1248/cpb.33.5083
Mata-Gómez L, Montañez JC, Méndez-Zavala A, Aguilar CN (2014) Biotechnological production
      of carotenoids by yeasts: An overview. Microbial Cell Factories 13:12. https://doi.
      org/10.1186/1475-2859-13-12
Ohsawa T, Yukawa M, Takao C, Murayama M, Bando H (1992) Studies on constituents of fruit body
      of Polyporus umbellatus and their cytotoxic activity. Chemical and Pharmaceutical Bulletin
      40:143−147. https://doi.org/10.1248/cpb.40.143
Palacios I, Lozano M, Moro C, D’Arrigo M, Rostagno M, Martínez, JA, García-Lafuente A, Guillamón
      E, Villares A (2011) Antioxidant properties of phenolic compounds occurring in edible
      mushroom. Food Chemistry 128:674−678. https://doi.org/10.1016/j.foodchem.2011.03.085
Pan X, Qin P, Liu R, Li J, Zhang F (2016) Molecular mechanism on two fluoroquinolones-induced
      oxidative stress: evidences from Copper/zinc Superoxide Dismutase. RSC Advances – The
      Royal Society of Chemistry 6:91141–91149. https://doi.org/10.1021/acs.jafc.8b02197
Pasailiuk MV (2019) Biological peculiarities of a rare mushroom Sparassis nemecii (Sparassidaceae,
      Polyporales) on plant substrates in pure culture. Ukrainian Journal of Botany 76:493−498.
      https://doi.org/10.15407/ukrbotj76.06.493
Pasailiuk MV (2020) Growing of Polyporus umbellatus. Current Research in Environmental & Applied
      Mycology (Journal of Fungal Biology) 10(1):457−465. https://doi.org/10.5943/cream/10/1/35
Piyarach K, Nipawan K, Chadapon C, Daluwan S, Kunjana R (2020) Effect of drying on β-carotene,
      α-carotene, lutein and zeaxanthin content in vegetables and its application for vegetable
      seasoning. E3S Web of Conferences 141:6. https://doi.org/10.1051/e3sconf/202014102007
Pleszczyńska M, Wiater A, Siwulski M, Szczodrak J (2012) Successful large-scale production of
      fruiting bodies of Laetiporus sulphureus (Bull.: Fr.) Murrill on an artificial substrate. World
      Journal of Microbiology & Biotechnology 29(4):753−758. https://doi.org/10.1007/s11274-
      012-1230-z
Podkowa A, Kryczyk-Poprawa A, Opoka W, Muszynska B (2021) Culinary–medicinal mushrooms:
      a review of organic compounds and bioelements with antioxidant activity. European Food
      Reserch and Technology 247:513–533. https://doi.org/10.1007/s00217-020-03646-1
Rahman MM, Islam MB, Biswas M, Alam AHMK (2015) In vitro antioxidant and free radical
      scavenging activity of different parts of Tabebuia pallida growing in Bangladesh. BMC
      Research Notes 8:621. https://doi.org/10.1186/s13104-015-1618-6
Ribeiro B, Pinho PG, Andrade PB, Oliveira C, César A, Ferreira S, Baptista P, Valentão P (2011) Do
      bioactive carotenoids contribute to the color of edible mushrooms? The Open Chemical and
      Biomedical Methods Journal 4:14-18. https://doi.org/10.2174/1875038901104010014
Robaszkiewicz A, Bartosz G, Lawrynowicz M, Soszyński M (2010) The role of Polyphenols,
      β-Carotene, and Lycopene in the antioxidative action of the extracts of dried, edible mushrooms.
      Journal of Nutrition and Metabolism 2010:173274. https://doi.org/10.1155/2010/173274
Scalbert A, Johnson IT, Saltmarsh M (2005) Polyphenols: antioxidants and beyond. The American
      Journal of Clinical Nutrition 81:215S–217S. https://doi.org/10.1093/ajcn/81.1.215S
Sułkowska-Ziaja K, Muszyńska B, Gawalska A, Sałaciak K (2018) Laetiporus sulphureus – chemical
      composition and medicinal value. Acta Scientarium Polonorum Hortorum Cultus 17(1):89–98.
      https://doi.org/10.24326/asphc.2018.1.8



                                                 124
Kopylchuk et al.                                             Italian Journal of Mycology 52 (2023): 112-125

https://doi.org/10.6092/issn.2531-7342/16457



Sun L, Liu Q, Bao C, Fan J (2017) Comparison of free total amino acid compositions and their
     functional classifications in 13 wild edible mushrooms. Molecules 24;22(3):350. https://doi.
     org/10.3390/molecules22030350
Turfan N, Ayan S, Pekşen A, Selin Akın S (2020) Antioxidant enzyme activities of some wild and
     cultivated edible mushrooms in Turkey. International Journal of Agriculture and Wildlife
     Science 6(2):202–209 https://doi.org/10.24180/ijaws.749512
Valadon LRG (1976) Carotenoids as additional taxonomic characters in fungi: a review. Transactions
     of the British Mycological Society. 67(1):1-15.
Wang Z, Binder M, Dai Y-C, Hibbett DS (2004) Phylogenetic rela-tionships of Sparassis inferred
     from nuclear and mitochondrialribosomal DNA and RNA polymerase sequences. Mycologia
     96:1015–1029. https://doi.org/10.1016/S0007-1536(76)80001-0
Wang Y, Wu B, Shao J, Jia J, Tian Y, Shu X, Ren X, Guan Y (2018) Extraction, purification and physico
     chemical properties of a novel lectin from Laetiporus sulphureus mushroom. Lebensmittel-
     Wissenschaft & Technologie 91:151−159. https://doi.org/10.1016/j.lwt.2018.01.032
Xing YM, Zhang LC, Liang HQ, Lv J, Song C, Guo SX, Wang CL, Lee TS, Lee MW (2013) Sclerotial
     formation of Polyporus umbellatus by low temperature treatment under artificial conditions.
     PLoS ONE 8:e56190. https://doi.org/10.1371/journal.pone.0056190
Yosioka I, Yamamoto T (1964) The constituents of chuling. 2-Hydroxytetracosanoic acid. Yakugaku
     Zasshi 84:742−744.
Yuwa-Amornpitak T, Butkhup L, Yeunyaw PN (2020) Amino acids and antioxidant activities
     of extracts from wild edible mushrooms from a community forest in the Nasrinual District,
     Maha Sarakham, Thailand. Food Science Technology (Campinas) 40(3):712‒720. https://doi.
     org/10.1590/fst.18519
Zhang Y, Lu X, Fu Z, Wang Z, Zhang J (2011) Sulphated modification of a polysaccharide obtained from
     fresh persimmon (Diospyros kaki L.) fruit and antioxidant activities of the sulphated derivatives.
     Food Chemistry 127(3):1084−1090. https://doi.org/10.1016/j.foodchem.2011.01.100
Zhang CC, Yin X, Cao CY, Wei J, Zhang Q, Gao JM (2015) Chemical constituents from Hericium
     erinaceus and their ability to stimulate NGF-mediated neurite outgrowth on PC12 cells.
     Bioorganic & Medicinal Chemistry Letters 25(22):5078−5082. https://doi.org/10.1016/j.
     bmcl.2015.10.016
Zhang CC, Cao CY, Kubo MH, Kenichi YXT, Fukuyama Y, Gao JM (2017) Chemical constituents
     from Hericium erinaceus promote neuronal survival and potentiate neurite outgrowth via
     the TrkA/Erk1/2 pathway. International Journal of Molecular Sciences 18:1659. https://doi.
     org/10.3390/ijms18081659
Zhang J, Wang F, Liu K, Liu Q, Yang Y, Dong C (2018) Heat and light stresses affect metabolite
     production in the fruit bodyof the medicinal mushroom Cordyceps militaris. Applied
     Microbiology and Biotechnology 102:4523–4533. https://doi.org/10.1007/s00253-018-8899-3
Zhao JD, Zhang XQ (1992) The polypores of China. J. Cramer, Berlin ‒ Stuttgart, Germany.
Zhou D, Ruan J, Cai Y, Xiong Z, Fu W, Wei A (2010) Antioxidant and hepatoprotective activity
     of ethanol extract of Arachniodes exilis (Hance) Ching. Journal of Ethnopharmacology
     010(129):232–237. https://doi.org/10.1016/j.jep.2010.03.016
Zhuang YL, Sun LP, Zhao X, Wang JF, Hou H, Li BF (2009) Antioxidant and melanogenesis-inhibitory
     activities of collagen peptide from Jellyfish (Rhopilema esculentum). Journal of the Science of
     Food and Agriculture 89:1722–1727. https://doi.org/10.1002/jsfa.3645




                                                 125
