Ergosterol Content and Antioxidant Activity of Lion’s Mane

Mushroom (Hericium erinaceus) and Its Induction to Vitamin D

by UVC-Irradiation

Pinida Joradon

, Vilai Rungsardthong

1,*

, Uracha Ruktanonchai

, Khomson Suttisintong

Tawin Iempridee

, Benjawan Thumthanaruk

, Savitri Vatanyoopaisarn

, Nutsuda Sumonsiri

and Dutsadee Uttapap

Department of Agro-Industrial, King Mongkut’s University of Technology North Bangkok, Bangkok, Thailand

National Nanotechnology Center, Thailand Science Park, Pathumthani, Thailand

Division of Biochemical Technology, King Mongkut’s University of Technology Thonburi, Bangkok, Thailand

Keywords: Lion’s Mane Mushroom (Hericium erinaceus), Ergosterol, Vitamin D

(Ergocalciferol), UVC, Antioxidant

Properties.

Abstract: Lion’s Mane mushroom (Hericium erinaceus), LM, is a medicinal mushroom which has high protein content

and contains many bioactive compounds. However, a large amount of the irregular-shape LM (Ir-LM),

considered as by-products, are generated during the cultivation. The objectives of this research were to

determine the ergosterol content in the LM and investigate the effect of ultraviolet (UV) irradiation on the

conversion of ergosterol in the Ir-LM extract to vitamin D

. Ir-LM extracts were investigated for its

antioxidant properties before dissolved in methanol and irradiated with UVC for 120 min at 5 cm distance

from the lamp. The results showed that the Ir-LM contained significantly higher (p < 0.05) ergosterol content

(2.52 ± 0.13 mg/g dried LM) than that of regular-shape LM (Reg-LM), 2.15 ± 0.08 mg/g dried LM. Ergosterol

at 1.74 ± 0.09 mg/g dried LM without vitamin D

was detected in the non-irradiated extract, while

interestingly, the irradiated sample showed a decrease of ergosterol at 13.5% with a detection of ergocalciferol

at 30.01 ± 7.09 µg/g dried LM. These obtained results exhibited a new area of post-extraction procedure

aiming to enhance vitamin D

enriched extracts from mushroom by-products which can be value-added as a

nutritional supplement in foods.

1 INTRODUCTION

Lion’s Mane mushroom (Hericium erinaceus, LM) is

an edible fungus which has been used in traditional

Chinese medicine for long time (Khan et al., 2013).

LM contains a significant content of bioactive

compounds, ergosterol, hericenone C and hericene A

(Joradon et al., 2022), that would might be

responsible for several health-promoting properties

(Friedman, 2015).

In general, high molecular weight substances

including polysaccharides and low molecular weight

substances including terpenoids can be used to

categorize bioactive metabolites in the LM (Thongbai

et al., 2015). The compounds with bioactivity exist in

different part of the mushroom (Shen et al., 2010), for

instance, low-molecular weight metabolites,

erinacines were found in mycelia while hericenones

were detected in fruiting bodies of LM (Thongbai et

al., 2015).

There is an increasing interest in bioactive

compounds from natural sources such as gamma

oryzanol from rice bran oil (Rodsuwan et al., 2020),

puerarin from Pueraria (Rungsardthong et al., 2021),

ergosterol from LM (Joradon et al., 2022;

Tachabenjarong et al., 2022), antioxidants from

Sacha Inchi oil (Suwannasang et al., 2021; 2022a;

2022b), and bioactive compounds from bamboo

mushroom (Binheam et al., 2022).

Ergosterol can exhibit anti-inflammatory, anti-

tyrosinase and anti-cancer properties (Kang et al.,

2015). According to Corrêa et al. (2017), ergosterol

has the potential to lessen the negative effects from

high cholesterol. Ergosterol is the most prevalent

sterol presents in the membranes of fungi, and it is

also a precursor of vitamin D

. The compound is

Joradon, P., Rungsardthong, V., Ruktanonchai, U., Suttisintong, K., Iempridee, T., Thumthanaruk, B., Vatanyoopaisarn, S., Sumonsiri, N. and Uttapap, D.

Ergosterol Content and Antioxidant Activity of Lion’s Mane Mushroom (Hericium erinaceus) and Its Induction to Vitamin D2 by UVC-Irradiation.

DOI: 10.5220/0011594600003430

In Proceedings of the 8th International Conference on Agricultural and Biological Sciences (ABS 2022), pages 19-28

ISBN: 978-989-758-607-1; ISSN: 2795-5893

important for maintaining fluidity and permeability,

involves with endocytosis and cytoskeletal

organization inside the fungal cells (Abe & Hiraki,

2009). Mushroom ergosterol can be converted into

vitamin D

(Jasinghe et al., 2007). With the exposure

to UV irradiation, ergosterol encounters

photochemical cleavage within their structure,

causing the formation of the intermediate of vitamin

. After being heated, this intermediate then goes

through thermal isomerization to create vitamin D

(Jasinghe et al., 2007).

One of the pro-hormones that is crucial for

maintaining human health is vitamin D. Vitamin D is

well known for supporting bone health and calcium

homeostasis, as well as having a variety of non-

skeletal effects on immunological function and cell

physiology (Durrant et al., 2022). Vitamin D

and D

are found as a major forms of vitamin D (Dawson-

Hughes et al., 2010). Typically, vitamin D

can be

found in the fruiting bodies of mushrooms. Applying

ultraviolet (UV) irradiation to the mushroom will lead

to the conversion of ergosterol to vitamin D

(Jäpelt

and Jakobsen, 2013). Morales et al. (2017) revealed

that UVC irradiation to the ethanol extract of Shiitake

mushroom (Lentinula edodes) at 25°C, for 1 h (4 cm

away from the light source) could generate a high

content of vitamin D

than direct irradiation of the

fruiting body.

During the cultivation of LM, irregular-shape LM

(Ir-LM) were found in the mushroom farm. These

mushrooms are considered as by-products and sold at

low prices in the mushroom market because of their

inferior morphology and quality. Therefore,

alternative solutions are required to increase the value

of these mushroom by-products which still contain a

high content of health-benefit compounds. They

might be used as high nutritional food or extracts

enriched with medicinal compounds for food or

medicinal uses. Consequently, the objectives of this

research were to determine the major bioactive

compounds, ergosterol, hericenone C and hericene A

contents and antioxidant properties in the extracts

prepared from regular LM (Reg-LM) and Ir-LM.

Further investigation on the effect of ultraviolet C

(UVC) irradiation on the conversion of ergosterol in

the Ir-LM extract to vitamin D

was carried out.

Morphology of fruiting bodies and their proximate

compositions were also performed. The results of this

study would propose an alternative way to increase

the value of the by-product by converting ergosterol

in the mushroom extracts to vitamin D

2 MATERIALS AND METHODS

2.1 Biological Materials and Chemicals

Fruiting bodies of Reg-LM and Ir-LM were

purchased from Fresh and Friendly Farm Co., Ltd. at

Thanyaburi district in Pathum Thani province,

Thailand. Morphology of the LM cultivated in the

farm for 4 lots were monitored and their production

yield and related economic data were calculated.

Ergosterol (95%) was obtained from Sigma-Aldrich

Química (Madrid, Spain). Ergocalciferol (98%)

(vitamin D

) was purchased from TCI, Japan. Folin-

Ciocalteu reagent and 2,2-diphenyl-1-

picreylhydrazyl (DPPH) were purchased from Sisco

Research Laboratories Pvt. Ltd., India while gallic

acid (98%) was the product from Sigma-Aldrich,

USA. Trolox reagent was obtained from M Tedia,

USA. Sodium carbonate (99.5% purity) was

purchased from Merck, India. Absolute ethanol

(analytical grade) was bought from PanReac

(Barcelona, Spain), while water and methanol (HPLC

grade) were purchased from LAB-SCAN (Gliwice,

Poland).

2.2 Cultivation of LM in the

Mushroom Farm

Temperature in the cultivation room was controlled at

16 ± 1 °C with the photo period at 12 hours of light

and 12 hours of darkness. The measurement of CO

in the room was measured by a sensor, and its average

intensity during the cultivation was around 900 mg/L.

Initial moisture content of the substrates for the

mycelium growth was 70-80%. Figure 1 shows the

substrate bags with LM mycelium growths in the

incubation room at Fresh and Friendly Farm.

2.3 Extraction and Determination of

Ergosterol

Ten grams of the freeze-dried samples from both

Reg-LM and Ir-LM were soaked in 200 mL of

absolute ethanol for 3 days at room temperature (25 ±

3 °C) in the dark. After filtering the suspensions, the

clean supernatant was collected and all solvents were

eliminated using a rotary evaporator (R114, Buchi,

Switzerland), at 50 °C. The dried extract was kept at

-20 °C in the dark until use. High-performance liquid

chromatography (HPLC) used to determine

ergosterol and vitamin D

in the dried extracts.

ABS 2022 - The International Conference on Agricultural and Biological Sciences

2.4 UVC-Irradiation of Ir-LM

The extract from Ir-LM was dissolved with absolute

methanol in cylindrical vessels and exposed to the

UVC at 254 nm with the intensity of 145 µw/cm

(determined by UVC meter, Solarmeter® version 8.0,

Solar Light Company Inc.) at room temperature for

120 min, at 5 cm away from the lamp. HPLC was

used to determine the levels of ergosterol and vitamin

2.5 Antioxidant Properties

2.5.1 Total Phenolic Content (TPC)

Total phenolic contents of the extracts from both Reg-

LM and Ir-LM were evaluated using Folin-Ciocalteu

reagent with the absorbance at 750 nm, measured by

a 96-well microplate reader (Bio-Rad, iMark, USA).

All experiments were performed in triplicates. The

TPC were determined as gallic acid equivalents

(GAE)/g dried LM using gallic acid as the reference

(Rosa et al., 2017).

2.5.2 DPPH Scavenging Activity

The DPPH radical scavenging experiment was

modified slightly from Ahmed et al. (2012) in order

to assess the antioxidant ability of LM extracts. DPPH

was dissolved in absolute methanol at a concentration

of 0.5 mM to create DPPH radical solution. Fifty µL

of DPPH solution were added to 50 µL of the extract

in each well. The plate was incubated for 30 minutes

in the dark at room temperature. Trolox was used to

create a calibration curve, with methanol serving as

the blank. Absorbance of the solution was measured

by a microplate reader (Bio-Rad, iMark, USA) at 540

nm. All experiments were carried out in triplicates

and scavenging ability was calculated as mg Trolox

equivalent (TE)/g dried LM followed Eq. 1:

% Inhibition = (























)×100

(1)

where A

control

is the absorbance of the control,

which has all reagents present minus the samples.

sample

is the absorbance of the mushroom extracts

with reagents added.

2.5.3 ABTS Scavenging Activity

The ABTS radical scavenging assay was performed

followed Ahmed et al. (2012) with minor

modification. The ABTS radical was determined by

reacting 200 mL of 140 mM K

solution with 7

mM of ABTS solution, and allowed the mixture to

react for 16 hours at room temperature in the dark.

Absorbance of the working ABTS solution was

measured on a microplate reader (Bio-Rad, iMark) at

750 nm. ABTS solution used for the measurement

was diluted with absolute methanol to gain the

absorbance approximately 1.1 to 1.2 at 750 nm. In 96-

well microtiter plates, 50 L of extract solution was

combined with 100 L of ABTS solution. Methanol

was used as a blank. The absorbance was read within

30 minutes at room temperature. All measurements

were performed in triplicate. The antioxidant activity

was determined as mg TE/g dried LM as detailed in

Eq. 1.

2.6 Analysis of Bioactive Compounds

by HPLC

Ergosterol and other compounds in the Reg-LM and

Ir-LM extracts were analyzed using HPLC (Agilent

Technology 1,200 series, Germany). Eclipse Zorbax

XDB-C 18 (Agilent, 250 4.6 mm, 5 m) analytical

column and Zorbax XDB-C 18 (Agilent, 12.5 4.6

mm, 5 m) guard column were used with the HPLC

system to measure the compounds at 282 nm using

the UVVIS LC detector. Methanol and water were

mixed in the mobile phase at a 98:2 ratio. Ergosterol

was quantified from the calibration curve of

ergosterol standard. The content of hericenone C and

hericene A were calculated from their peak area

compared with the area of ergosterol.

The content of each bioactive compound after

extraction was presented in the unit of mg/g extract,

mg/g dried LM, and mg/g fresh LM. The unit mg/g

extract was calculated as mg bioactive compound per

gram of dried extract while mg/g dried LM and mg/g

fresh LM were calculated as the mg bioactive

compound per gram of dried LM powder and fresh

LM fruiting body, respectively.

2.7 Statistical Data Analysis

All experiments were performed with three

replications. Data were analyzed using IBM SPSS 26

for Windows (SPSS Inc.) with independent t-test to

compare the means of each treatment. To compare the

difference between each sample, the significance

level at p < 0.05 was employed.

Ergosterol Content and Antioxidant Activity of Lion’s Mane Mushroom (Hericium erinaceus) and Its Induction to Vitamin D2 by

UVC-Irradiation

3 RESULTS AND DISCUSSIONS

3.1 Cultivation of LM in the

Mushroom Farm

Production yield, percentage of Reg-LM and Ir-LM

as well as the calculated economic losses were

presented in Table 1. Irregular-shape LM (Ir-LM)

occurred during the cultivation were sold as low

price, consequently leads to economic loss for the

mushroom farm. Ir-LM is considered as by-products,

equals to around 20% of total LM cultivated in the

farm. Total production of LM in the farm was about

3,270 Bahts/batch or 156,967 Bahts/year since

approximately 48 batches were cultivated per year

(Table 1). Similar information was reported by

Aguayo et al. (2017) that high amounts of irregular-

shape of Button mushroom (Agaricus Bisporus)

fruiting bodies (around 20% of total production) were

generated during the mushroom cultivation. They are

considered as by-products since their misshaped caps

or stalks did not meet the product specifications set

by retailers.

Table 1: Statistical data for the production of Lion’s Mane

mushroom at Fresh & Friendly Farm mushroom farm

-LM I

-LM

Production yield (Kg/batch)

168

(80%)

25 (20%)

Sale amount (Bahts/batch) 109,473 13,081

Economic loss (Bahts/batch) - 3,270

Economic loss (Bahts/year) - 156,967

Notes:

Data were collected and averaged from 4 batches of

the cultivation during February, 2022.

Irregular-shape LM was found with the covered

lids, randomly in the incubation room for mycelium

and primordium induction. Most of the Ir-LM was

found at the top of the shelf in the incubation room

(Figure 1).

Figure 1: Incubation for the mycelium growths of Lion’s

Mane mushroom in the substrate bags at Fresh and Friendly

Farm. The temperature in the incubation room was

controlled at 16 ± 1 °C.

Morphological study indicated that the fruiting

bodies of Reg-LM exhibited long spore-bearing

spines and intricately branches with primary (PB),

secondary (SB) and tertiary (TB) branches (Figure 2,

A-C), while the fruiting bodies of Ir-LM aggregated

thickly in branches which presented only primary

(PB) and secondary (SB) branches with short spore-

bearing spines (Figure 2, D-F).

Figure 2: Macroscopic images on the fruiting bodies of

Lion’s Mane mushroom. (A-C): regular-shape (Reg-LM),

and (D-F): irregular-shape (Ir-LM) mushroom. SP: spore-

bearing spine, PB: primary branch, SB: second branch, TB:

tertiary branch.

There are various environmental factors affecting

the formation of primordia and fruiting bodies. The

occurrence of Ir-LM might be due to both intrinsic

and extrinsic factors. The spawning rate and the

synthesis of volatile organic molecules like ethylene

and 1-octen-3-ol may be examples of intrinsic factors.

Yang et al. (2013) reported that an increase of

spawning rate could reduce the time for mycelial

colonization development of the fruiting bodies.

According to Zhang et al. (2016), reducing the

generation of ethylene and its precursor 1-

aminocyclopropane-1-carboxylic acid (ACC)

resulted in twice as many primordia that developed

more quickly than those in the wild type strain of the

button mushroom. Eastwood et al. (2013), reported

that temperature and the reduced content of octenol

(1-octen-3-ol) were the switches that controlled the

plenary morphogenesis process, led to the production

of fruiting bodies from the vegetative mycelium.

Before directing the formation of undifferentiated

hyphae into mature mushrooms, the volatile 1-octen-

3-ol would first affect the differentiation of mycelial

condenses into hyphal knots. Undifferentiated

primordia would subsequently become differentiating

primordia, depending on the temperature (Baars et al.,

2020).

The possible extrinsic factors such as air

composition and luminosity in the room might

involve with the occurrence of Ir-LM since large

amounts of Ir-LM were found at the top of the shelf

ABS 2022 - The International Conference on Agricultural and Biological Sciences

in the incubation room. According to Bellettini et al.

(2019), mushroom fruiting body in a cultivation room

might be generated by reducing the amounts of

carbon dioxide and the rate of air circulation. The

spawn bottles on the top shelf might expose to higher

light intensity and air ventilation than the other shelf

at lower levels (Figure 1). The higher light intensity

in the environment caused the reduction of coloration,

deformations and elongated stipe of mushroom

fruiting bodies (Urben, 2004). In order to promote pin

head production in button mushrooms, Visscher et al.

(1979) hypothesized that an optimal concentration of

ethylene and carbon dioxide are required.

3.2 Extraction and Determination of

Ergosterol, Hericenone C and

Hericene A

The ergosterol content shown in Table 2 reveals that

Ir-LM contained significantly higher (p < 0.05)

ergosterol content (2.52 ± 0.13 mg/g dried LM) than

that of Reg-LM (2.15 ± 0.08 mg/g dried LM).

Gąsecka et al. (2020) reported that the sample

preparation techniques had a substantial impact on the

ergosterol content of the LM mushroom.

Specifically,

the authors noted that ergosterol level was higher in

the fresh mushroom samples (4.5 mg ergosterol/kg

LM), and it declined when drying temperature was

increased from 20 to 70 °C. Heleno et al. (2016)

presented the use of ethanol as the extraction solvent

in a soxhlet extraction of button mushrooms for 4

hours, yielding maximum concentration of ergosterol

at 676 ± 3 mg/100 g. Mushroom by-products can be

used for the recovery of bioactive compound from

fruiting bodies or mycelium of mushrooms. Wang et

al. (2015) found antichronic atrophic gastritis activity

from the mycelium of LM by-products extracted with

hot water (70 °C for 12 h) followed by precipitation

with ethanol (80%). The antioxidant and antifungal

activities in the ultrasound-assisted extract of LM

mycelium with ethyl acetate were reported by Lu et

al. (2014). Ergosterol was also extracted from the by-

product of button mushroom fruiting bodies using

ethanol and microwave-assisted at 132.8 °C and 1.6

g/L CO

flow rate for 19.4 min (Heleno et al., 2016).

Several chromatogram peaks were detected in

both LM extracts, Reg-LM and Ir-LM and the

contents of hericenone C and hericene A were

compared (Figure 3, B-C). Ir-LM extract showed

significantly higher (p < 0.05) in hericene A (1.25 ±

0.08 mg/g extract) than Reg-LM extracts (0.42 ± 0.02

mg/g extract) (Table 3). One of the bioactive

substances in LM was hericene A, which had an IC

value of 6.7 M and significantly reduced glucosidase

activity (Lee at al., 2020). However, the content of

hericenone C in both Reg-LM and Ir-LM were not

significantly different. Ergosterol, hericenone C, and

hericene A in the fresh Ir-LM fruiting bodies were

found at 0.49 ± 0.03 mg/g fresh LM, 0.11 ± 0.01 mg/g

fresh LM, and 0.02 ± 0.01 mg/g fresh LM,

respectively.

Table 2: Ergosterol content in the extracts prepared from

regular-shape (Reg-LM) and irregular-shape (Ir-LM)

Lion’s Mane mushroom.

Extract

Ergosterol

mg/g extract

mg/g dried

mg/g fresh

Reg-

19.42

±0.66



2.15

±0.08



0.41

±0.02



Ir-LM

26.77

±1.20



2.52

±0.13



0.49

±0.02



Notes: Different superscripts in the same column mean

significant difference at p < 0.05.

Table 3: Concentration of the hericenone C and hericene A in regular-shape (Reg-LM) and irregular-shape (Ir-LM) Lion’s

Mane mushroom.

Extract

Hericenone C

Hericene A

mg/g extract

mg/g dried

mg/g fresh

mg/g extract

mg/g dried

mg/g fresh LM

Reg-LM

5.40 ± 0.18



0.60 ± 0.02



0.12 ± 0.01



0.42 ± 0.02



0.05 ± 0.01



0.01 ± 0.01



Ir-LM

5.79 ± 0.36



1.25 ± 0.08



0.11 ± 0.01



0.55 ± 0.04



0.12 ± 0.01



0.02 ± 0.01



Notes: Different superscripts in the same column mean significant difference at p < 0.05.

The content of hericenone C and

hericene A were calculated from their peak area compared with that area of ergosterol.

3.3 Antioxidant Properties

Reg-LM and Ir-LM extracts were tested for their

capacity to inhibit the DPPH radical, one of the few

stable organic nitrogen radicals that presents purple

color. This test relies on the determination of DPPH

•

loss upon sample response. Additionally, the

antioxidant activity in the ABTS experiment was

Ergosterol Content and Antioxidant Activity of Lion’s Mane Mushroom (Hericium erinaceus) and Its Induction to Vitamin D2 by

UVC-Irradiation

calculated as the capacity of the extract to reduce

color when they have direct contact with the radical

ABTS

•+

(Rivero-Cruz et al., 2020). DPPH and ABTS

scavenging ability for Ir-LM extract were 0.27 ± 0.01

mg TE/g dried LM and 0.52 ± 0.04 mg TE/g dried

LM, respectively as presented in Table 4. These

antioxidant activities of Ir-LM were significantly

higher (p < 0.05) than Reg-LM extract which

presented 0.20 ± 0.01 mg TE/g dried LM and 0.46 ±

0.01 mg TE/g dried LM of DPPH and ABTS

scavenging ability, respectively. The antioxidant

concentration required needed to reduce the initial

radical concentration by 50% is known as IC

, which

is a factor commonly used to evaluate antioxidant

activity (Rivero-Cruz et al., 2020). The IC

determined by DPPH (85.28 mg/ml), and ABTS

scavenging ability (164.84 mg/ml) of Reg-LM

extracts were higher than those of Ir-LM extract,

67.03 mg/ml and 151.27 mg/ml, respectively. In

conclusion, Ir-LM exhibited higher antioxidant

activities in terms of DPPH and ABTS than Reg-LM.

The reasons that Ir-LM expressed significantly higher

antioxidant capacity might be because it contained

higher phenolic content (0.11 ± 0.02 mg GAE/g dried

LM) than that of Reg-LM (0.06 ± 0.01mg GAE/g

dried LM). Phenolic acids are important compounds

contributing to antioxidant activity due to OH groups

that can scavenge free radicals are present in their

structures (Heleno et al., 2012).

(1) Ergosterol and vitamin D

standard

(2) Bioactive compounds extracted from LM by Maceration (3) Ergosterol and vitamin D

before and after UVC irradiation

Figure 3: HPLC chromatograms of ergosterol and vitamin D

standard (A), crude ethanolic extract of regular-shape Lion’s

Mane mushroom (B), irregular-shape Lion’s Mane mushroom (C), non-irradiated Ir-LM extract (D), and Ir-LM extract

irradiated with UV-C (E).

Moreover, the higher ergosterol content in Ir-

LM (Table 2) might involve with its antioxidant

capacity. Dupont et al. (2021) described that the B-

ring of ergosterol has two double bonds, which may

Ergosterol

Vitamin D



Hericenone C

Hericene A

Ergosterol

Vitamin D



Hericenone C

Hericene A

Ergosterol

Standard

Reg-LM

extract

Ir-LM

extract

Non-irradiated

Ir-LM extract

Irradiated

Ir-LM

extract

No peak of

Vitamin D



ABS 2022 - The International Conference on Agricultural and Biological Sciences

have antioxidant effects. Shao et al. (2010) also

observed that, ergosterol was primarily responsible

for the antioxidant activity in the lipophilic fraction

of button mushroom. Therefore, Ir-LM was selected

to be used for ultraviolet C (UVC) irradiation to

convert ergosterol to vitamin D

in the extract.

3.4 Induction of Vitamin D

by UVC

Irradiation

The most abundant sterol in cell membranes of fungi

is ergosterol. It is critical for preserving permeability,

trafficking, fluidity, and cytoskeletal structure (Abe

and Hiraki, 2009). The transformation of ergosterol to

vitamin D

can be obtained by the use of UV

radiation, either artificially or naturally (Jäpelt and

Jakobsen, 2013). Ergosterol undergoes

photochemical cleavage at the B ring upon exposure

to UV radiation, resulting in the synthesis of pre-

vitamin D

, an intermediate of vitamin D

. After

being heated, this intermediate then goes through

thermal isomerization to produce vitamin D

. The

equilibrium between thermal and photochemical

processes is crucial for the production of vitamin D

(Jasinghe et al., 2007).

The UV radiation for food processing and

preservation is effective and favorable for the

environment (Singh et al., 2021). Food irradiation is

a technology that is secure and effective. The flavor

of the product, taste, and odor, is unaffected by the

radiation, and neither are the residues or poisons

generated in the process (Bisht et al., 2021).

Table 4: Radical scavenging ability and total phenolic

content in the extracts from the fruiting bodies of regular-

shape (Reg-LM) and irregular-shape (Ir-LM) Lion’s Mane

mushroom.

-LM I

-LM

Total phenolic content

(mg GAE/g dried LM)

0.06

±0.01



0.11

±0.02



DPPH radical scavenging

ability (mg TE /g dried LM)

0.20

±0.01



0.27

±0.01





(mg/ml) by DPPH

85.28 67.03

ABTS radical scavenging

ability (mg TE /g dried LM)

0.46

±0.01



0.52

±0.04





(mg/ml) by ABTS

164.84 151.27

Notes: Different superscripts in the same row mean

significant difference at p < 0.05. IC

is the concentration

of antioxidants required to decrease the initial radical

concentration by 50%; DPPH: 2,2-diphenyl-1-

picrylhydrazyl, ABTS: 3-ethylbenzthiazoline-6-sulphonic

acid.

Vitamin D (D

, or D

, or both) ingested in the

human body is incorporated into chylomicrons, which

are absorbed by the lymphatic system and penetrated

into venous blood. Vitamin D

cannot be

biosynthesized by human body. Most oil-rich fish

including the oil fish from salmon contains high

content of vitamin D

. Vitamin D sufficiency can

enhance the absorption of calcium and phosphorus by

30-40% and 80%, respectively (Nair & Maseeh,

2012). According to the World Health Organization,

low vitamin D intake, both in the form of D

and D

causes bone problems and raises the risk of other

chronic diseases (Holick & Chen, 2008). Ergosterol

was detected in the non-irradiated extract at 1.74 ±

0.09 mg/g dried LM, but no vitamin D

was found.

Interestingly, a decrease of ergosterol at 13.5% with

a detection of vitamin D

at 30.01 ± 7.09 µg/g dried

LM were found in the irradiated extract (Table 5).

Table 5: Ergosterol and vitamin D

content in the extracts

of non-irradiated and irradiated Lion’s Mane mushroom

with UV-C at 5 cm distance from UV-C lamp, for 2 h.

Samples

Ergosterol

(mg/g dried

LM)

Vitamin D



(µg/g dried LM)

Non-irradiated

extract

(Control)

1.74 ± 0.09



Irradiated

extract

1.51 ± 0.15



30.01 ±7.09

Notes: ns: no significant difference at p < 0.05.

Different superscripts in the same column mean significant

difference at p < 0.05. nd: not detected

In addition, a decrease of ergosterol content was

observed in the irradiated extract. Ergosterol might be

transformed into other derivatives of vitamin D such

as lumisterol or tachysterol (Morales et al., 2017).

There are very few studies on the process of directly

exposing mushroom extract to UVC to convert

ergosterol to vitamin D

. Morales et al. (2017)

revealed that UVC irradiation to Shiitake mushroom

extracts in ethanol at 25°C, for 1 h, at 4 cm distance

from the lamp could generate vitamin D

enriched

extracts higher than the irradiation of direct fruiting

body. Generally, vitamin D

was induced by direct

UV-light irradiation to fresh fruiting bodies or dried

mushroom powder. Xu et al. (2020) reported that

after being exposed to a high level of UVC (4 kJ/m

)

for 40 minutes, vitamin D

content of ground shiitake

and Jew's ear powder increased from 1.38 g/g to 20.11

g/g and 4.13 g/g to 39.93 g/g, respectively.

With a UVB lamp at 25 °C, for 2 hours, and 19

cm away from the lamp, Huang et al. (2015) were able

to irradiate oyster mushrooms and obtain 69 g/g

vitamin D. In addition, Wittig et al. (2013) exposed

Ergosterol Content and Antioxidant Activity of Lion’s Mane Mushroom (Hericium erinaceus) and Its Induction to Vitamin D2 by

UVC-Irradiation

the same mushrooms to UVB radiation at 20 and 30

°C with a 10 cm distance from the light and found that

after only 10 minutes of exposure, a higher vitamin D

content (80 g/g) was obtained. These studies present

that the mushrooms placed more closely to the UV

lamp could generate a higher amount of vitamin D.

Currently, several industrial mushroom farms in

the United States, Ireland, the Netherlands, and

Australia have exposed their fresh mushrooms to UV

light, producing at least 10 µg of vitamin D per 100 g

of fresh weight. Therefore, a 100 g serving of the

mushroom can satisfy 50 to 100% of a person's daily

requirement for vitamin D. Additionally, UV-light-

exposed dry mushrooms can also create sufficient

content of vitamin D

for nutritional purposes

(Cardwell et al., 2018).

4 CONCLUSIONS

Irregular-shape Lion’s Mane (Ir-LM), mushroom

considered as by-products, was used in the

experiment to investigate the effect of UVC

irradiation on the conversion of ergosterol in the

mushroom extract to vitamin D

. Ergosterol,

hericenone C, hericene A, total phenolic content and

antioxidant activities of Ir-LM were found

significantly higher (p < 0.05) than those of Reg-LM.

Irradiation with a low dose of UVC (145 µW/cm

) for

120 minutes at 5 cm distance from the lamp caused

the detection of vitamin D

in the irradiated extracts,

but no detection in the non-irradiated sample. This

research provides possible methods for a conversion

to vitamin D

enriched extracts from mushroom by-

products and use as nutritional supplement in

medicinal foods. It is necessary to conduct additional

research on the negative effects of UVC irradiation on

the antioxidant activity, physical characteristics, and

other significant nutritional parameters of irradiated

mushroom extracts. The study on the effect of

individual environmental factors on the induction of

Ir-LM fruiting bodies are required.

ACKNOWLEDGEMENTS

This research was funded by King Mongkut’s

University of Technology North Bangkok and

National Science and Technology Development

Agency, Thailand (Contract no. 024/2563). This

research was also partially supported by NRCT

Senior Research Scholar Program (Contract No.814-

2020). Special thanks go to Mr. Wuttipong

Ruksavong, Managing Director, Fresh and Friendly

Farm Co., Ltd., for his kind supports during the

cultivation of the Lion’s Mane mushrooms.

REFERENCES

Abe, F., and Hiraki, T. (2009). Mechanistic role of

ergosterol in membrane rigidity and cycloheximide

resistance in Saccharomyces cerevisiae. Biochimica et

Biophysica Acta (BBA)-Biomembranes, 1788:743-752.

Aguayo, I. A., Walton, J., Vinas, I. and Tiwarii, B. K.,

(2017). Ultrasound assisted extraction of

polysaccharides from mushroom by-products. LWT -

Food Science and Technology, 77:92e99.

Ahmed, A. S., Elgorashi, E. E., Moodley, N., McGaw, L.

J., Naidoo, V. and Eloff, J. N. (2012). The

antimicrobial, antioxidative, anti-inflammatory activity

and cytotoxicity of different fractions of four South

African Bauhinia species used traditionally to treat

diarrhoea, Journal of Ethnopharmacology, 143:826-

839.

Baars, J. J. P., Scholtmeijer, K., Sonnenberg, A. S. M. and

Peer, A. V. (2020). Critical factors involved in

primordia building in agaricus bisporus: a review.

Molecules, 25:2984.

Bellettini, M. B., Assump¸ F., Fiorda, A., Maieves, H. A.,

Teixeira, G. L., A´vila, S., Hornung, P. S., Ju´nior, A.

M. and Ribani, R. H. (2019). Factors affecting

mushroom Pleurotus spp. Saudi Journal of Biological

Sciences, 26:633-646.

Binheam, F., Thumthanaruk, B., Vatanyoopaisarn, S.,

Rungsardthong, V. and Poodchakarn, S. (2022).

Antioxidant capacity of the bamboo mushroom

(Dictyophora indusiata) and the effects of temperature

and relative humidity on its growth. In IAMBEST 2022,

3rd International Conference on Informatics,

Agriculture, Management, Business administration,

Engineering, Science and Technology.

Bisht, B., Bhatnagar, P., Gururani, P., Kumar, V., Tomarf,

M. S., Sinhmar, R., Rathi, N. and Kumar, S. (2021).

Food irradiation: effect of ionizing and non-ionizing

radiations on preservation of fruits and vegetables-a

review. Trends in Food Science & Technology, 114:

372-385.

Cardwell, G., Bornman, J. F. James, A.P., and Black L.J.

(2018). A review of mushrooms as a potential source of

dietary vitamin D. Nutrients, 10: 1498.

doi:10.3390/nu10101498

Corrêa, R. C. G., Peralta, R. M., Bracht, A. and Ferreira, I.

C. F. R. (2017). The emerging use of mycosterols in

food industry along with the current trend of extended

use of bioactive phytosterols. Trends in Food Science

& Technology, 67:19-35.

Dawson-Hughes, B., Mithal, A., Bonjour, J. P., Boonen, S.,

Burckhardt, P., Fuleihan, G. E. H., Josse, R. G., Lips,

P., Morales-Torres, J. L. A. and Yoshimura, N. (2010).

IOF position statement: vitamin D recommendations

ABS 2022 - The International Conference on Agricultural and Biological Sciences

for older adults. Osteoporosis International, 21: 1151-

1154.

Dupont, S., Fleurat-Lessard, P., Cruz, R.G, Lafarge, C.,

Grangeteau, C., Yahou, F., Gerbeau-Pissot, P., Júnior,

O.A., Gervais, P., Simon-Plas, F., Cayot, P. and Beney,

L. (2021). Antioxidant properties of ergosterol and its

role in yeast resistance to oxidation. Antioxidants, 10:

1024.

Durrant, L. R., Bucca, G., Hesketh, A., Möller-Levet, C.,

Tripkovic, L., Wu, H., Hart, K. H., Mathers, J. C.,

Elliott, R. M., Lanham-New, S. A. and Smith, C. P.

(2022). Vitamins D

and D

have overlapping but

different effects on the human immune system revealed

through analysis of the blood transcriptome. Frontiers

in Immunology, 13:790444.

Eastwood, D. C., Herman, B., Noble, R., Dobrovin-

Pennington, A., Sreenivasaprasad, S. and Burton, K. S.

(2013). Environmental regulation of reproductive phase

change in Agaricus bisporus by 1-octen-3-ol,

temperature and CO

. Fungal Genetics and Biology,

55:54-66.

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.

Gasecka, M., Siwulski, M., Magdziak, Z., Budzyn´ska, S.,

Stuper-Szablewska, K., Niedzielski, P. and Mleczek,

M. (2020). The effect of drying temperature on

bioactive compounds and antioxidant activity of

Leccinum scabrum (Bull.) Gray and Hericium

erinaceus (Bull.) Journal of Food Science and

Technology, 57(2):513-525.

Heleno, S. A., Barros, L., Martins, A., Queiroz, M. J. R. P.,

Santos-Buelga, C. and Ferreira, I. C. F. R. (2012).

Phenolic, polysaccharidic, and lipidic fractions of

mushrooms from North eastern Portugal: chemical

compounds with antioxidant properties. Journal of

Agricultural and Food Chemistry, 60:4634-4640.

Heleno, S. A., Prieto, M. A., Barros, L., Rodrigues, A.A.,

Barreiro, M.F. and Ferreira, I.C.F.R. (2016).

Optimization of microwave-assisted extraction of

ergosterol from Agaricus bisporus L. by-products using

response surface methodology. Food and Bioproducts

Processing, 100:25-35.

Holick, M. F. and Chen, T. C. (2008). Vitamin D

deficiency: A worldwide problem with health

consequences. The American Journal of Clinical

Nutrition,87:1080-1086.

Huang, S. J., Lin, C. P. and Tsai, S. Y. (2015). Vitamin D

content and antioxidant properties of fruit body and

mycelia of edible mushrooms by UV-B irradiation.

Journal of Food Composition and Analysis, 42:38-45.

Jäpelt, R. B. and Jakobsen, J. (2013). Vitamin D in plants:

A review of occurrence, analysis, and biosynthesis.

Frontiers of Plant Science, 4:136-136.

Jasinghe, V. J., Perera, C. O. and Sablani, S. S. (2007).

Kinetics of the conversion of ergosterol in edible

mushrooms. Journal of Food Engineering, 79(3):864-

869.

Joradon, P., Rungsardthong, V., Ruktanonchai, U.,

Suttisintong, K., Iempridee, T., Thumthanaruk, B.,

Vatanyoopaisarn, S. and Uttapap, D. (2022). A

comparative study of conventional and supercritical

carbon dioxide extraction methods for the recovery of

bioactive compounds from Lion’s Mane mushroom

(Hericium erinaceus). In RI

C 2022, International

Conference on Research, Invention, and Innovation

Congress.

Kang, J. H., Jang, J. E., Mishra, S. K., Lee, H. J., Nho, C.

W., Shin, D., Jin, M., Kim, M. K., Choi, C. and Oh, S.

H. (2015). Ergosterol peroxide from Chaga mushroom

(Inonotus obliquus) exhibits anti-cancer activity by

down-regulation of the β-catenin pathway in colorectal

cancer. Journal of Ethnopharmacology, 173:303-312.

Khan, M. A., Tania, M., Liu, R. and Rahman, M. M. (2013).

Hericium erinaceus: an edible mushroom with

medicinal values. Journal of Complementary and

Integrative Medicine, 24(10).

Lee, S.K., Ryu, S.H., Turk, A., Yeon, S.W., Jo, Y.H., Han,

Y.K., Hwang, B.Y., Lee, K.Y. and Lee, M.K. (2020).

Characterization of α-glucosidase inhibitory

constituents of the fruiting body of lion’s Mane

mushroom (Hericium erinaceus), Journal of

Ethnopharmacology, 262:113197.

Lu, Q. Q., Tian, J. M., Wei, J. and Gao, J. M. (2014).

Bioactive metabolites from the mycelia of the

basidiomycete Hericium erinaceum. Natural Product

Research, 28:288-1292.

Morales, D., Gil-Ramirez, A., Smiderle, F. R., Piris, A. J.,

Ruiz-Rodriguez, A. and Rivasa, C. (2017). Vitamin D-

enriched extracts obtained from shiitake mushrooms

(Lentinula edodes) by supercritical fluid extraction and

UV-irradiation. Innovative Food Science and Emerging

Technologies, 41:330-336.

Nair, R. and Maseeh, A. (2012). Vitamin D: The “sunshine”

vitamin. Journal of Pharmacology and

Pharmacotherapeutics, 3(2).

Rodsuwan, U., Pithanthanakul, U., Thisayakorn, K.,

Uttapap, D., Boonpisuttinant, K., Vatanyoopaisarn, S.,

Thumthanaruk, B. and Rungsardthong, B. (2020).

Preparation and characterization of gamma oryzanol

loaded zein nanoparticles and its improved stability.

Food Science and Nutrition, 9(2):616-624.

Rosa, A., Maxia, A., Putzu, D., Atzeri, A., Era, B., Fais, A.,

Sanna, C. and Piras, A. (2017). Chemical composition

of Lycium europaeum fruit oil obtained by supercritical

extraction and evaluation of its antioxidant

activity, cytotoxicity and cell absorption. Food

Chemistry, 230:82-9.

Rivero-Cruz, J. F., Granados-Pineda, J., Pedraza-Chaverri,

J., Pérez-Rojas, J. M., Kumar-Passari, A., Diaz-Ruiz,

G. and Rivero-Cruz, B. E. (2020). Phytochemical

Constituents, antioxidant, cytotoxic, and antimicrobial

activities of the ethanolic extract of Mexican Brown

propolis. Antioxidants, 9(70).

Rungsardthong, V., Pithanthanakul, U., Puttanlek, C.,

Uttapap, D. and Boonpisuttinant, K. (2021).

Preparation of puerarin-loaded zein nanoparticles:

Ergosterol Content and Antioxidant Activity of Lion’s Mane Mushroom (Hericium erinaceus) and Its Induction to Vitamin D2 by

UVC-Irradiation

characterization and stability study. Journal of Current

Science and Technology, 11(1):60-70.

Shao, S., Hernandez, M., Kramer, J. K., Rinker, D. L. and

Tsao, R. (2010). Ergosterol profiles, fatty acid

composition, and antioxidant activities of button

mushrooms as affected by tissue part and

developmental stage. Journal of Agricultural and Food

Chemistry, 58:11616-11625.

Shen, J. W., Yu, H. Y., Ruan, Y., Wu, T. T. and Zhao, X.

(2010). Hericenones and erinacines: stimulators of

nerve growth factor (NGF) biosynthesis in Hericium

erinaceus. Mycology An International Journal on

Fungal Biology, 1:92-98.

Singh, H., Bhardwaj, S. K., Khatri, M., Kim, K. H. and

Bhardwaj, N. (2021). UVC radiation for food safety:

An emerging technology for the microbial disinfection

of food products. Chemical Engineering Journal, 417:

128084.

Suwannasang, S., Thumthanaruk, B., Zhong, Q., Uttapap,

D., Puttanlek, C., Vatanyoopaisarn, S. and

Rungsardthong, V. (2021). The improved properties of

zein encapsulating and stabilizing Sacha Inchi oil by

surfactant combination of lecithin and Tween 80. Food

and Bioprocess Technology, 14(11):2078-2090.

Suwannasang, S., Zhong, Q., Thumthanaruk, B.,

Vatanyoopaisarn, S., Uttapap, D., Puttanlek, C. and

Rungsardthong, V. (2022a). Physicochemical

properties of yogurt fortified with microencapsulated

Sacha Inchi oil. LWT, 61:113375.

Suwannasang, S., Zhong, Q., Thumthanaruk, B.,

Vatanyoopaisarn, S., Uttapap, D., Puttanlek, C. and

Rungsardthong, V. (2022b). Optimization of wall

material composition for production of spray-dried

Sacha Inchi oil microcapsules with desirable

physicochemical properties. Food and Bioprocess

Technology. (Major revision)

Tachabenjarong, N., Rungsardthong, V., Ruktanonchi, U.,

Suttisintong, K., lempridee, T., Thumthanaruk, B.,

Vatanyoopaisam, S., Poodchakarn, S. and Uttapap, D.

(2022). Bioactive compound and antioxidant activity of

Lion’s mane mushroom (Hericium erinaceus) from

different growth period. In RI

C 2022, International

Conference on Research, Invention, and Innovation

Congress.

Thongbai, B., Rapior, S., Hyde, K. D., Wittstein, K. and

Stadler, M. (2015). Hericium erinaceus, an amazing

medicinal mushroom. Mycological Progress, 14(91).

Visscher, H. R. (1979). Fructification of Agaricus bisporus

(Lge) Imb. in relation to the relevant microflora in the

casing soil. Mushroom Science, 10:631-663.

Urben, A. F. (2004). Produc¸ao de cogumelos por meio de

tecnologia chinesa modificada. Embrapa Recursos

Gene´ticose Biotecnologia, Brasılia (in Portuguese).

Wang, M., Gao, Y., Xu, D. and Gao, Q. (2015). A

polysaccharide from cultured mycelium of Hericium

erinaceus and its anti-chronic atrophic gastritis activity.

International Journal of Biological Macromolecules,

81:656-661.

Wittig, M., Krings, U. and Berger, R. G. (2013). Single-run

analysis of vitamin D photoproducts in oyster

mushroom (Pleurotus ostreatus) after UV-B treatment.

Journal of Food Composition and Analysis, 31:266-

274.

Xu, Z., Meenu, M. and Xu, B. (2020). Effects of UV-C

treatment and ultrafine-grinding on the

biotransformation of ergosterol to vitamin D

physiochemical properties, and antioxidant properties

of shiitake and Jew’s ear. Food Chemistry, 309:125738.

Yang, W., Guo, F., Wan, Z., and Saudi, J. (2013). Yield and

size of oyster mushroom grown on rice/wheat straw

basal substrate supplemented with cotton seed hull.

Journal of Biological Sciences, 20:333-338.

Zhang, C., Huang, T., Shen, C., Wang, X., Qi, Y., Shen, J.,

Song, A., Qiu, L. and Ai, Y. (2016). Downregulation of

ethylene production increases mycelial growth and

primordia formation in the button culinary-medicinal

mushroom, Agaricus bisporus (agaricomycetes).

International Journal of Medicinal Mushrooms, 18:

1131-1140.

ABS 2022 - The International Conference on Agricultural and Biological Sciences