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Antimicrobial Effect of the Lingzhi or Reishi Medicinal Mushroom, Ganoderma

lucidum (Higher Basidiomycetes) and Its Main Compounds

Article in International Journal of Medicinal Mushrooms · June 2014

DOI: 10.1615/IntJMedMushr.v16.i1.70

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 International Journal of Medicinal Mushrooms, 16(1): 77–84 (2014)

Antimicrobial Effect of the Lingzhi or Reishi

Medicinal Mushroom, Ganoderma lucidum (Higher
Basidiomycetes) and Its Main Compounds
Mahdi Vazirian,1 Mohammad Ali Faramarzi,2 Seyed Esmaeil Sadat Ebrahimi,3 Hamid Reza
Monsef Esfahani,1 Nasrin Samadi,4 Seyed Aboulfazl Hosseini,1 Ali Asghari,1 Azadeh Manayi,1 Ali
Mousazadeh,5 Mohammad Reza Asef,6 Emran Habibi,1 & Yaghoub Amanzadeh1,*
1
Department of Pharmacognosy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran;
2
Department of Pharmaceutical Biotechnology, Faculty of Pharmacy and Biotechnology Research Center, Tehran
University of Medical Sciences, Tehran, Iran; 3Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran
University of Medical Sciences, Tehran, Iran; 4Department of Drug and Food Control, Faculty of Pharmacy and
Pharmaceutical Quality Assurance Research Center, Tehran University of Medical Sciences, Tehran, Iran; 5Agriculture
and Natural Research Center of Mazandaran, Passand Forest and Rangeland Research Station, Behshahr, Iran;
6
Department of Botany, Iranian Research Institute of Plant Protection, Tehran, Iran

*Address all correspondence to: Yaghoub Amanzadeh, Department of Pharmacognosy, Faculty of Pharmacy, Tehran University of Medical
Sciences, Enqelab Ave., Tehran 14155-6451, Iran; Tel: +98-21-66954706; [email protected].

ABSTRACT: Mushrooms are considered one of the richest sources of natural antibiotics, and various species of them
inhibit the growth of a wide diversity of microorganisms. Ganoderma lucidum, a well-known medicinal mushroom.
has many pharmacological and biological activities including an antimicrobial effect, although few studies have
investigated the antibacterial and antifungal effects of its purified compounds. The chemical structure of the purified
compounds from the hexane fraction was elucidated as ergosta-7,22-dien-3β-yl acetate, ergosta-5,7,22-trien-3β-yl
acetate (isopyrocalciferol acetate), ergosta-7,22-dien-3-one, ergosta-7,22-dien-3β-ol, and ergosta-5,7,22-trien-3β-
ol (ergostrol). In addition, the structure of ganodermadiol was demonstrated after purification from the chloroform
fraction. The fractions inhibited Gram-positive bacteria and yeast, with minimum inhibitory concentration values
of 6.25 mg/mL, but were ineffective against Gram-negative bacteria in the tested concentrations. The results were
comparable for isolated compounds, whereas the mixture of ergosta-7,22-dien-3β-yl acetate and isopyrocalciferol
acetate was weakly effective against Escherichia coli (minimum inhibitory concentration, 10 mg/mL). It could
be assumed that the antimicrobial effect of crude fractions is the consequence of mixing triterpenoid and steroid
compounds.

KEY WORDS: medicinal mushrooms, Ganoderma lucidum, nonpolar fractions, steroids, triterpenoids, antimicrobial
effect

ABBREVIATIONS: ATCC, American Type Culture Collection; 13C, carbon-13; dd, doublet of doublets; 1H,
hydrogen-1; MIC, minimum inhibitory concentration; NMR, muclear magnetic resonance; TLC, thin layer
chromatography; UV, ultraviolet.

I. INTRODUCTION rally occurring components from herbs and medici-

nal mushrooms, mainly terpenes, steroids, saponins,
Natural products have long attracted a great deal of flavones, xanthones, coumarins, and alkaloids, are
interest among researchers in isolation of antibac- reported to have antimicrobial effects.3
terial agents.1 This is mainly due to the spread of Mushrooms have been considered a rich source
resistance among microorganisms and the toxicity of natural antibiotics, and various species inhibit the
of synthetic/semisynthetic medications that are in growth of a wide diversity of microorganisms. For
many instances major hindrances to gaining suc- instance, some species of mushrooms produce sev-
cessful therapeutic outcomes.2 A number of natu- eral bioactive metabolites that have potential me-

1045-4403/14/$35.00 © 2014 Begell House, Inc. www.begellhouse.com 77

78 Vazirian et al.

dicinal attributes.3 The Lingzhi or Reishi medicinal lin.15 The crude alcoholic and purified extracts of
mushroom Ganoderma lucidum (W. Curt.:Fr.) P. G. lucidum exhibited various degrees of inhibition
Karst. (Ganodermataceae, higher Basidiomycetes), against tested Gram-negative and Gram-positive
a wood-degrading species, is a well-known medic- bacteria and some fungi.16
inal mushroom; more than 300 reports concern its This study was undertaken to elucidate the
chemical constituents and related species.4 Among antibacterial and antifungal effects of hexane and
different bioactive compounds of this mushroom chloroform fractions of G. lucidum and isolated
(including triterpenoids, sterols, polysaccharides, compounds against 2 Gram-positive bacteria (S.
and fatty acids),5 triterpenoids and polysaccharides aureus and B. subtilis), 2 Gram-negative bacteria
are considered the main pharmacologically active (Pseudomonas aeruginosa and Escherichia coli),
constituents and are reported to possess many ben- and one yeast (Candida albicans) using the disc
eficial effects.2,6–8 diffusion method. The antimicrobial effect of iso-
A survey of earlier studies about medici- lated compounds was evaluated here for the first
nal mushrooms, including Ganoderma species, time.
revealed that there are few studies about the an-
timicrobial effect of their purified compounds. II. MATERIALS AND METHODS
Evaluation of the antimicrobial effect of purified
triterpenoids and steroids demonstrated that la- A. Mushroom Material
nostane-type terpenoids, purified from the ethyl
acetate extract of G. lucidum, exhibited in vitro G. lucidum fruit bodies were collected in the
antiplasmodial activity.9 Polar and nonpolar ex- Neka forests (Mazandaran Province, Iran) from
tracts of 4 species of Ganoderma tested with direct the stumps of Carpinus betulus as its host in May
bioautography on thin layer chromatography were 2011, and a voucher specimen (Iran 15697 F) was
active against both Gram-positive and Gram-neg- deposited in the herbarium of the Iranian Research
ative strains.10 Institute of Plant Protection.
The aqueous extract of G. lucidum has inhib-
ited to different extents 15 types of Gram-positive B. Instruments for Chemical Analysis
and Gram-negative bacteria; the most potent in-
hibition occurs against Micrococcus luteus (mini- Column chromatography was carried out using sil-
mum inhibitory concentratino [MIC], 0.75 mg/ ica gel (70–230 mesh; Merck) and Sephadex LH-
mL).2 In another work, acetone extract of the 20 (Fluka) as stationary phase. Precoated silica gel
mushroom showed the highest antibacterial activ- 60 F254 plates (Merck) were used for thin layer
ity against Klebsiella pneumoniae (MIC, 4.33 ± chromatography (TLC). Spots on TLC plates were
0.33 mg/mL), whereas another study revealed that detected under ultraviolet (UV) light at 254 and
G. lucidum methanol extract possessed efficient 366 nm using a UV/Vis Cecil CE 1021 spectro-
antibacterial effect on methicillin-resistant Staphy- photometer and visualized by spraying the devel-
lococcus aureus (MRSA) and Bacillus subtilis.11–13 oped plates with anisaldehyde followed by heating
The results of another study indicated that for 5 minutes. Nuclear magnetic resonance (NMR)
extract of G. lucidum favorably showed remark- spectra were obtained on a Bruker DRX 500 (500
able antibacterial activity against Gram-positive MHz for hydrogen-1 [1H] NMR and 125 MHz for
bacteria but only a slight effect on Gram-negative carbon-13 [13C] NMR), with tetramethylsilane
strains.14 Further studies demonstrated that extract used as the internal standard. Infrared spectra were
of the Reishi mushroom resulted in an additive and recorded on a Nicolet 550-A Spectrometer on so-
synergist effect in combination with 4 antibiotics, dium chloride pellets.
with a synergistic activity against B. subtilis and
Klebsiella oxytoca when combined with cefazo-

International Journal of Medicinal Mushrooms

Antimicrobial Effect of Ganoderma lucidum 79

C. Extraction, Isolation, and Structure tion [ATCC] 6538) and B. subtilis (ATCC 6633);
Elucidation 2 Gram-negative bacteria: P. aeruginosa (ATCC
9027) and E. coli (ATCC 8739); and a yeast: C.
The fruit bodies of the mushrooms were air dried albicans (ATCC 10231). The bacteria inocula were
and grounded into small pieces. We subsequently prepared by suspending colonies from Mueller-
extracted 800 g of ground mushroom using hex- Hinton agar media in 0.9% saline overnight. The
ane and chloroform, each for 72 hours three times. C. albicans inocula were prepared by suspend-
Solvents were distilled before use. The samples ing colonies from 48 to 72 hours old Sabouraud
treated with the hexane and chloroform extracts dextrose agar cultures in 0.9% saline. The inocula
were dried using a rotary evaporator to give 12.3 were adjusted photometrically at 600 nm to a cell
and 13.5 g dried extracts, respectively. The hex- density equivalent to 0.5 McFarland standard (1.5
ane fraction was subjected to column chromatog- × 108 colony-forming units/mL). Mueller-Hinton
raphy on silica gel eluted with hexane/ethyl acetate and Sabouraud dextrose agar plates (100-mm di-
(9:1–0:10) to afford 8 subfractions (H1–H9). On the ameter) were seeded individually with bacterial
base of analytical TLC, H3 (20 mg) was applied on or fungal suspensions using a sterile cotton swab.
a silica gel column and eluted with hexane/ethyl Fractions and compounds were dissolved in chlo-
acetate (9:1–7:3), yielding a 1:1 mixture (12 mg) roform at concentrations of 100–3.125 mg/mL and
of compounds 1 (ergosta-7,22-dien-3β-yl acetate) 10–1.25 mg/mL, respectively. A blank sterile disc
and 2 (ergosta-5,7,22-trien-3β-yl acetate). The H6 (6 mm in diameter) impregnated with known con-
fraction (30 mg) was applied to silica gel and elut- centrations of extracts and compounds (which were
ed with hexane/ethyl acetate (9:1–7:3), yielding screened for antibacterial properties at 10 mg/mL)
compound 3 (ergosta-7,22-dien-3-one; 9.5 mg). was placed gently onto the agar. The plates with the
The H9 fraction (102 mg) was purified by silica bacteria and the yeast were incubated at 37°C for
gel and eluted with hexane/ethyl acetate (6:4–3:7) 24 hours and 20–25°C for 48 hours, respectively.
to give a 1:1 mixture (10.8 mg) of compounds 4 The tests were done in triplicate. After incubation,
(ergosta-7,22-dien-3β-ol) and 5 (ergosta-5,7,22- the mean inhibition zone diameter (in millimeters)
trien-3β-ol). The chloroform fraction was puri- at the tested concentration (10 mg/mL) was deter-
fied through application to silica gel and elution mined and the lowest concentration with a zone of
using hexane/ethyl acetate (9:1–5:5), affording 9 inhibition considered the MIC.
new fractions (C1–C9). Subfraction C9 (68 mg) was
applied to a silica gel using hexane/ethyl acetate III. RESULTS AND DISCUSSION
(6:4–2:8) and then rechromatographed on a Sepha-
dex LH-20 eluted with chloroform/methanol (5:5) After successive chromatography separations, the
to give compound 6 (ganodermadiol; 19.5 mg). hexane fraction (efficiency, 1.54% w/w) afforded
compounds 1–5. In addition, the chloroform frac-
D. Antimicrobial Test tion (efficiency, 1.69% w/w) provided compound
6 (Fig. 1). The following shows the spectrometric
Sabouraud dextrose agar (Merck) and Mueller- data of the compounds.
Hinton agar (Merck) were purchased for the anti- • Compound 1 (ergosta-7,22-dien-3β-yl ac-
microbial tests. The UV/Vis Cecil CE 1021 spec- etate): 13C-NMR (500 MHz, CDCl3) δ 37.08
trophotometer was used again in the microbial tests (CH2, C-1), 31.91 (CH2, C-2), 72.46 (CH, C-3),
for preparation of 0.5 McFarland standards. Using 37.91 (CH2, C-4), 40.04 (CH, C-5), 29.68 (C,
the disc diffusion method,11,12 we determined the C-6), 117.31 (CH, C-7), 139.48 (C, C-8), 49.24
antimicrobial effects of fractions and semipurified/ (CH, C-9), 34.20 (C, C-10), 21.45 (CH2, C-11),
purified compounds against 2 Gram-positive bac- 39.36 (CH2, C-12), 43.25 (C, C-13), 55.03 (CH,
teria: S. aureus (American Type Culture Collec- C-14), 22.90 (CH2, C-15), 28.09 (CH2, C-16),

Volume 16, Number 1, 2014

80 Vazirian et al.

FIG. 1: Structures of the purified compounds from G. lucidum. 1: ergosta-7,22-dien-3β-yl acetate; 2: ergosta-5,7,22-
trien-3β-yl acetate (isopyrocalciferol acetate); 3: ergosta-7,22-dien-3-one; 4: ergosta-7,22-dien-3β-ol; 5: ergosta-
5,7,22-trien-3β-ol (ergosterol); and 6: ganodermadiol.

55.67 (CH, C-17), 12.08 (CH3, C-18), 12.94 C-11), 39.00 (CH2, C-12), 43.25 (C, C-13), 54.50
(CH3, C-19), 40.43 (CH, C-20), 19.63 (CH3, (CH, C-14), 22.97 (CH2, C-15), 28.27 (CH2,C-16),
C-21), 131.85 (CH, C-22), 135.55 (CH, C-23), 55.90 (CH, C-17), 12.04 (CH3, C-18), 16.16 (CH3,
42.79 (CH, C-24), 33.07 (CH, C-25), 19.94 (CH3, C-19), 40.43 (CH, C-20), 19.63 (CH3, C-21),
C-26), 21.09 (CH3, C-27), 17.59 (CH3, C-28), 131.95 (CH, C-22), 135.65 (CH, C-23), 42.79
22.68 (CH3, acetate). (CH, C-24), 33.07 (CH, C-25), 19.94 (CH3, C-26),
• Compound 2 (ergosta-5,7,22-trien-3β-yl ac- 21.09 (CH3, C-27), 17.59 (CH3, C-28), 22.68 (CH3,
etate): 1H-NMR (500 MHz, CDCl3) δ 5.58 (1H, acetate).
doublet of doublets [dd], H-6), 3.6 (1H, m, H-3), • Compound 3 (ergosta-7,22-dien-3-one): In-
1.03 (3H, d, J = 6.6 Hz, H-21), 0.95 (3H, singlet, frared (sodium chloride): 2952, 2925, 1716, 1450,
H-19), 0.92 (3H, d, J=6.7 Hz, H-26), 0.64 (3H, 1376, 1240 cm-1; 1H-NMR (500 MHz, CDCl3) δ
singlet, H-18); 13C-NMR (500 MHz, CDCl3) δ 5.19 (1H, dd, J = 7.15 Hz, H-22), 5.22 (1H, dd,
39.00 (CH2, C-1), 31.91 (CH2, C-2), 73.13 (CH, J = 7.15 Hz, H-23), 5.2 (1H, broad singlet, H-7),
C-3), 40.49 (CH2, C-4), 141.48 (C, C-5), 120.12 1.03 (3H, d, J = 6.8 Hz, H-21), 1.02 (3H, singlet,
(CH, C-6), 116.28 (CH, C-7), 139.48 (C, C-8), H-19), 0.92 (3H, doublet, J = 6.8 Hz, H-26), 0.85
46.01 (CH, C-9), 36.83 (C, C-10), 21.00 (CH2, (3H, doublet, J = 7.2 Hz, H-28), 0.83 (3H, dou-

International Journal of Medicinal Mushrooms

Antimicrobial Effect of Ganoderma lucidum 81

blet, J = 7.2 Hz, H-27), 0.58 (3H, singlet, H-18); • Compound 6 (ganodermadiol): 1H-NMR (500
13
C-NMR (500 MHz, CDCl3) δ 38.75 (CH2, C-1), MHz, CDCl3) δ 5.51 (1H, broad doublet, J = 6.1 Hz,
38.11 (CH2, C-2), 212.03 (C, C-3), 44.23 (CH2, H-7), 5.41 (1H, triplet, J = 6.4 Hz, H-24), 5.32 (1H,
C-4), 42.79 (CH, C-5), 30.03 (C, C-6), 116.98 dd, J = 6 Hz, H-11), 4.01 (2H, singlet, H-26), 3.2
(CH, C-7), 139.51 (C, C-8), 48.82 (CH, C-9), 34.4 (1H, dd, J = 4.4 and 11.4 Hz, H-3), 1.68 (3H, singlet,
(C, C-10), 21.67 (CH2, C-11), 39.39 (CH2, C-12), H-27), 1.03 (3H, singlet, H-30), 0.99 (3H, singlet,
43.26 (C, C-13), 54.95 (CH, C-14), 22.89 (CH2, H-19), 0.93 (3H, doublet, J = 6.4 Hz, H-21), 0.89
C-15), 28.08 (CH2,C-16), 55.9 (CH, C-17), 12.13 (3H, singlet, H-28), 0.89 (3H, singlet, H-29), 0.57
(CH3, C-18), 12.45 (CH3, C-19), 40.47 (CH, C-20), (3H, singlet, H-18); 13C-NMR (500 MHz, CDCl3) δ
21.1 (CH3, C-21), 135.56 (CH, C-22), 131.96 35.69 (C-1), 28.13 (C-2), 78.95 (C-3), 38.69 (C-4),
(CH, C-23), 42.85 (CH, C-24), 33.06 (CH, C-25), 50.3 (C-5), 23.42 (C-6), 120.21 (C-7), 142.64 (C-
17.58 (CH3, C-26), 19.63 (CH3, C-27), 19.93 (CH3, 8), 145.89 (CH, C-9), 37.35 (C-10), 116.26 (C-11),
C-28). 37.8 (C-12), 43.75 (C-13), 49.09 (C-14), 27.9 (C-
• Compound 4 (ergosta-7,22-dien-3β-ol): 1H- 15), 31.49 (C-16), 50.9 (C-17), 15.64 (CH3, C-18),
NMR (500 MHz, CDCl3) δ 3.6 (1H, multiplet, 22.74 (C-19), 36.07 (C-20), 18.39 (C-21), 35.91 (C-
H-3), 1.04 (3H, doublet, J = 6.4 Hz, H-21), 0.85 22), 24.52 (C-23), 126.99 (C-24), 134.33 (C-25),
(3H, singlet, H-19), 0.93 (3H, doublet, J = 6.7 Hz, 69.09 (C-26), 13.64 (C-27), 25.55 (C-28), 27.79 (C-
H-26), 0.54 (3H, singlet, H-18); 13C-NMR (500 29), 15.78 (C-30).
MHz, CDCl3) δ 37.11 (CH2, C-1), 31.43 (CH2, Structures of the purified compounds were
C-2), 71.05 (CH, C-3), 37.94 (CH2, C-4), 40.23 elucidated on the basis of comparison with those
(CH, C-5), 29.61 (C, C-6), 117.44 (CH, C-7), found in the literature. We identified 5 steroids
139.55 (C, C-8), 49.41 (CH, C-9), 34.19 (C, C-10), from hexane (1:1 mixture of compounds 1 and 2)17
21.52 (CH2, C-11), 39.42 (CH2, C-12), 43.28 (C, , compound 3,18 and a 1:1 mixture of compounds 4
C-13), 55.08 (CH, C-14), 22.91 (CH2, C-15), 28.1 and 5,17 followed by a triterpenoid (compound 6)19
(CH2,C-16), 55.69 (CH, C-17), 12.07 (CH3, C-18), from of the chloroform fraction.
13.03 (CH3,C-19), 40.43 (CH, C-20), 19.63 (CH3, As presented in Table 1, MIC values and mean
C-21), 131.84 (CH, C-22), 135.54 (CH, C-23), inhibition zones of each sample were assessed.
42.79 (CH, C-24), 33.07 (CH, C-25), 19.94 (CH3, The nonpolar fractions of G. lucidum supressed the
C-26), 21.09 (CH3, C-27), 17.59 (CH3, C-28). growth of Gram-positive bacteria and C. albicans
• Compound 5 (ergosta-5,7,22-trien-3β-ol): with an MIC value of 6.25 mg/mL. Most isolated
1
H-NMR (500 MHz, CDCl3) δ 5.58 (1H, dd, H-6), compounds (3–6) inhibited the growth of bacte-
3.6 (1H, m, H-3), 1.03 (3H, doublet J = 6.6 Hz, ria at higher concentrations than did the extracts
H-21), 0.95 (3H, singlet, H-19), 0.92 (3H, dou- (MIC, 5 mg/mL), whereas S. aureus was inhibited
blet, J = 6.7 Hz, H-26), 0.64 (3H, singlet, H-18); by a lower concentration of compounds 1 and 2
13
C-NMR (500 MHz, CDCl3) δ 38.35 (CH2, C-1), (MIC, 2.5 mg/mL). The latter also inhibited the
31.95 (CH2, C-2), 70.44 (CH, C-3), 40.75 (CH2, growth of E. coli at an MIC of 10 mg/mL. Howev-
C-4), 141.37 (C, C-5), 119.56 (CH, C-6), 116.25 er, the mixture of compounds 1 and 2 was ineffec-
(CH, C-7), 139.76 (C, C-8), 46.21 (CH, C-9), 37 tive against the yeast at tested concentrations. The
(C, C-10), 21.52 (CH2, C-11), 39.05 (CH2, C-12), minimum inhibition zone of both tested extracts
43.28 (C, C-13), 54.53 (CH, C-14), 22.97 (CH2, were evaluated in range of 16–17.2 mm for Gram-
C-15), 28.28 (CH2, C-16), 55.92 (CH, C-17), 12.03 positive bacteria and yeast, whereas the zone of in-
(CH3, C-18), 16.26 (CH3, C-19), 40.43 (CH, C-20), hibition for semipurified/purified compounds was
19.63 (CH3, C-21), 131.94 (CH, C-22), 135.66 measured in the range of 6.7–10.5 mm against S.
(CH, C-23), 42.79 (CH, C-24), 33.07 (CH, C-25), aureus, B. subtilis, and C. albicans.
19.94 (CH3, C-26), 21.09 (CH3, C-27), 17.59 (CH3, Antibacterial activity of G. lucidum extract
C-28). against Gram-positive strains has been observed

Volume 16, Number 1, 2014

82 Vazirian et al.

TABLE 1: Minimum Inhibitory Concentrations and Zone of Inhibitions of Fractions and Purified Com-
pounds from Ganoderma lucidum
Com-
Chloroform Hexane frac- pounds 1 Com- Compounds Com-
fraction tion and 2 pound 3 4 and 5 pound 6
MIC/ MIC/ MIC/ MIC/ MIC/ MIC/
Microorganism ZI ZI ZI ZI ZI ZI
Staphylococcus 6.25/ 6.25/ 2.5/ 5/ 5/ 5/
aureus 17.2 ± 0.5 17.2 ± 0.8 10.3 ± 0.1 9.7 ± 0.3 9.7±0.6 9.7 ± 0.8
Bacillus subtilis 6.25/ 6.25/ 5/ 5/ 5/ 5/
17.6 ± 0.6 17.0 ± 0.5 9.9 ± 0.1 9.6 ± 0.7 9.5±0.5 9.6 ± 0.3
Escherichia coli >10/ >10/ 10/ >10/ >10/ >10/
– – 6.9 ± 0.1 – – –
Pseudomonas >10/ >10/ >10/ >10/ >10/ >10/
aeruginosa – – – – – –
Candida albicans 6.25/ 6.25/ – 5/ 5/ 5/
16.6 ± 0.03 16.3 ± 0.3 9.6 ± 0.9 9.6 ± 0.1 9.6 ± 0.8
Data are shown as minimum inhibitory concentrations (mg/mL)/zone of inhibitions (millimeters ± standard
deviation) tested at a concentration of 10 mg/mL. Compounds 1 and 2 comprise a 1:1 mixture of ergosta-
5,7,22-trien-3β-yl acetate and ergosta-5,7,22-dien-3β-yl acetate. Compound 3 is ergosta-7,22-dien-3-one;
compounds 4 and 5 comprise a 1:1 mixture of ergosta-7,22-dien-3β-ol and ergosta-5,7,22-trien,3β-ol; and
compound 6 is ganodermadiol.

previously,20 and this effect has been related to 0.003–2.0 mg/mL; minimum bactericidal concen-
water-soluble ingredients and triterpenoids, which tration, 0.06–4.0 mg/mL) than the Gram-negative
could inhibit Gram-positive and/or Gram-nega- bacteria (MIC, 1.0–4.0 mg/mL; minimum bacte-
tive bacteria in other studies.2 Smania et al.21 has ricidal concentration, 2.0–4.0 mg/mL).2,22 Some
examined the antimicrobial effect of 3 steroids, previous results showed a weak inhibitory effect
including 5α-ergost-7-en-3β-ol, 5α-ergosta-7,22- of different extracts of G. lucidum against micro-
dien-3β-ol, and 5,8-epidioxy-5α,8α-ergosta-6,22- organisms, especially Gram-positive bacteria and
dien-3β-ol, and 5 triterpenes (applanoxidic ac- fungi. The acetone extract of the mushroom has
ids A, C, F, G, and H isolated from Ganoderma been effective against K. pneumoniae, with an
anulare) against 2 fungi, Microsporum canis and MIC of 4.33 ± 0.33 mg/mL.11
Trichophyton mentagrophytes. The applanoxidic In this study, among purified compounds,
acids A, C, and F were found to inhibit the growth ganodermadiol (compound 6) has previously
of the fungi at concentrations of 0.5 to 1 mg/mL.21 been examined for antiviral effect and possesses
In another study, crude alcoholic extract of G. lu- in vitro antiviral activity against influenza virus
cidum has been effective against Gram-negative type A. Furthermore, this compound was active
bacteria (MIC, 7–11.3 mg/mL), Gram-positive against herpes simplex virus type 1.22 Likewise,
bacteria (MIC, 10.3–11.0 mg/mL), and fungi 5α-ergosta-7,22-dien-3β-ol (compound 4) has
(MIC, 5–16.8 mg/mL). been evaluated for its antifungal effect against Mi-
Two steroids (i.e., 5α-ergosta-7en-3β-ol and crococcus canis and Trichophyton mentagrophytes
5,8-epidioxy-5α,8α-ergosta-6,22-dien-3β-ol) (MICs >1000 µg/mL).21 In another study, after
isolated from the fruit body of Ganoderma ap- extraction and purification from dichloromethane
planatum exhibited inhibitory activity against extract, this compound did not show any inhibitory
Gram-positive and Gram-negative bacteria; the effect against B. subtilis within tested concentra-
Gram-positive bacteria were more sensitive (MIC, tions (≥50 µg/mL).23

International Journal of Medicinal Mushrooms

Antimicrobial Effect of Ganoderma lucidum 83

The results of our study indicate that non- sible involvement of immunomodulating effects of
polar fractions and their semipurified/purified Ganoderma spp. on antimicrobial activity,2 other
compounds, including ergostan-type steroids and experiments must be incorporated in the evalua-
one lanostan-type triterpeoid, can inhibit Gram- tion of antimicrobial effect, maybe by examining
positive bacteria, and the tested concentrations of the use of Ganoderma in biologic systems.
some of them (compounds 3, 4, 5, and 6) also in-
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