Cytotoxic naphthoquinone and a new succinate ester from the soil fungus Fusarium solani PSU-RSPG227

Article abstract of DOI:10.1016/j.phytol.2014.11.018

A new naphthoquinone, solaninaphthoquione (1), and a new succinate ester derivative, 4-(4-hydroxyphenethoxy)-4-oxobutanoic acid (2), were isolated from the soil fungus Fusarium solani PSU-RSPG227 together with five previously reported compounds; javanicin (3), monaspilosin (4), aspergillol B (5), tyrosol (6) and 4-hydroxyphenylacetic acid (7). Their structures were elucidated primarily by NMR spectroscopic data. Due to paucity of materials, compounds 2, 4 and 5 as well as analogues of 5 were prepared for biological activity evaluation. Compound 1 showed significant cytotoxic activity against breast cancer (MCF-7) cells and mild cytotoxic activity against oral human carcinoma (KB) cells (IC₅₀ values of 21.3 and 22.6 μM, respectively) compared to standard compounds. Compound 1 also displayed weak antimalarial activity (IC₅₀ of 26.1 μM).

Full text of DOI:10.1016/j.phytol.2014.11.018

G Model
PHYTOL 837 1–5
Phytochemistry Letters xxx (2014) xxx–xxx
Contents lists available at ScienceDirect
Phytochemistry Letters
journal homepage: www.elsevier.com/locate/phytol
1
2
3
Cytotoxic naphthoquinone and a new succinate ester from the soil
fungus Fusarium solani PSU-RSPG227
a
a
a
Q1 Kwanruthai Tadpetch a, , Chatsuda Chukong , Laksamee Jeanmard , Aticha Thiraporn ,
*
4
5
Vatcharin Rukachaisirikul a, Souwalak Phongpaichit b, Jariya Sakayaroj c
6
7
8
9
a Department of Chemistry and Center of Excellence for Innnovation in Chemistry, Faculty of Science, Prince of Songkla University, Hat Yai,
Songkhla 90112, Thailand
b Department of Microbiology, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand
c National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand Science Park, Klong Luang, Pathumthani 12120, Thailand
A R T I C L E I N F O
A B S T R A C T
new naphthoquinone, solaninaphthoquione (1), and
Article history:
A
a new succinate ester derivative, 4-(4-
Received 13 August 2014
Received in revised form 24 November 2014
Accepted 27 November 2014
Available online xxx
hydroxyphenethoxy)-4-oxobutanoic acid (2), were isolated from the soil fungus Fusarium solani PSU-
RSPG227 together with five previously reported compounds; javanicin (3), monaspilosin (4), aspergillol
B (5), tyrosol (6) and 4-hydroxyphenylacetic acid (7). Their structures were elucidated primarily by NMR
spectroscopic data. Due to paucity of materials, compounds 2, 4 and 5 as well as analogues of 5 were
prepared for biological activity evaluation. Compound 1 showed significant cytotoxic activity against
breast cancer (MCF-7) cells and mild cytotoxic activity against oral human carcinoma (KB) cells (IC50
Keywords:
Fusarium solani
Naphthoquinone
Succinate ester
Synthesis
values of 21.3 and 22.6
m
M, respectively) compared to standard compounds. Compound 1 also displayed
weak antimalarial activity (IC50 of 26.1
mM).
ß 2014 Published by Elsevier B.V. on behalf of Phytochemical Society of Europe.
Cytotoxic activity
Antimalarial activity
10
11
1. Introduction
(Cheng et al., 2008), aspergillol B (5) (Pettit et al., 2009; Wang et al., 28
2009), tyrosol (6) (Rukachaisirikul et al., 2010) and 4-hydro- 29
xyphenylacetic acid (7) (Bala Show Reddy and Dhananjaya, 2000) 30
were isolated from the ethyl acetate extract of the culture broth of 31
F. solani PSU-RSPG227 (Fig. 1). In this Letter, we describe the 32
isolation and structure elucidation of 1 and 2 as well as the 33
syntheses of 2, 4, 5 and other derivatives and their biological 34
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
Q2
Fungi of the genus Fusarium have been recognized as one of
important sources of secondary metabolites with diverse biologi-
cal activities (Booth, 1971; Vesonder and Golinski, 1989; Nelson
et al., 1994; Bra¨se et al., 2009). Selected examples of this group
Q3 of metabolites include cytotoxic zearalenone (Stob et al., 1962),
antifungal fusapyrone (Altomare et al., 2000), antibacterial
fusaruside (Shu et al., 2004) and antimalarial apicidin F (von
Bargen et al., 2013). As part of ongoing search for biologically active
compounds from fungi, we investigated the fungus Fusarium solani
PSU-RSPG227 isolated from a soil sample collected at the Plant
Genetic Conservation Project under the Royal Initiative of Her
Royal Highness Princess Maha Chakri Sirindhorn, Rajjaprabha
Dam, Surat Thani Province, Thailand. Two new metabolites,
solaninaphthoquinone (1) and 4-(4-hydroxyphenethoxy)-4-oxo-
butanoic acid (2), together with five previously reported com-
pounds: javanicin (3) (Kimura et al., 1988), monaspilosin (4)
activity studies.
35
2. Results and discussion
36
Compound 1 was obtained as an orange amorphous solid. Its 37
molecular formula C15H14O5 was established by HREIMS (m/z 38
274.0836 [M+], calcd 274.0841) which corresponded to 9 degrees 39
of unsaturation and was in accordance with 15 signals of non- 40
equivalent carbons in the 13C NMR spectrum. The UV (lmax 221 41
and 268 nm), IR (1606 cmꢀ1) and 13C NMR (dC 182.9 and 189.7) 42
data suggested the presence of a 1,4-quinone moiety (Medentsev 43
and Akimenko, 1998). The IR spectrum also showed the absorption 44
bands for hydroxyl and ketone carbonyl groups at 3152 and 45
1716 cmꢀ1, respectively. The 1H NMR and 13C NMR spectroscopic 46
data of 1 are summarized in Table 1. The 1H NMR spectrum 47
consisted of signals corresponding to one chelated hydroxyl proton 48
*
Corresponding author. Tel.: +66 74 288437; fax: +66 74 558841.
E-mail addresses: kwanruthai.t@psu.ac.th, kwanruthai@hotmail.com
(K. Tadpetch).
http://dx.doi.org/10.1016/j.phytol.2014.11.018
1874-3900/ß 2014 Published by Elsevier B.V. on behalf of Phytochemical Society of Europe.
Please cite this article in press as: Tadpetch, K., et al., Cytotoxic naphthoquinone and a new succinate ester from the soil fungus Fusarium
solani PSU-RSPG227. Phytochem. Lett. (2014), http://dx.doi.org/10.1016/j.phytol.2014.11.018

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PHYTOL 837 1–5
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K. Tadpetch et al. / Phytochemistry Letters xxx (2014) xxx–xxx
Fig. 1. Structures of compounds 1–7.
49
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56
57
58
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60
61
62
63
64
65
66
67
68
69
70
71
(
dH 12.41, s), two ortho-coupled aromatic protons (dH 7.68, d,
peaks to C-4a, C-7 and C-8 in the HMBC spectrum. The other two
72
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75
76
77
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80
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103
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108
109
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111
112
113
114
115
J = 8.4 Hz and H 7.08, d, J = 8.4 Hz), one methoxyl group ( H 3.99),
d
d
substituents, the methyl and the 2-oxopropyl groups, would be on
ring A. The 2-oxopropyl unit was assigned at C-2 based on the HMBC
correlations of H2-10 to C-1 in addition to C-2, C-3 and C-11. This
assignment would place the methyl group at C-3, which was in
agreementwithHMBC correlations of the methyl protons to C-2, C-3
andC-4. Consequently, solaninaphthoquinone(1)wasidentifiedasa
new 1,4-naphthoquinone derivative.
Compound 2 was isolated as a white solid with molecular
formula C12H14O5 and six degrees of unsaturation as determined by
HREIMS (m/z 238.0846 [M+], calcd 238.0841). The IR spectrum
suggested the presence of hydroxyl (3270 cmꢀ1) and carbonyl
(1721 cmꢀ1) groups. The 1H NMR spectrum showed signals for two
one aromatic methyl group (dH 2.13) and a 2-oxopropyl unit (dH
3.80, s, 2H and dH 2.13, s, 3H). The presence of signals for ortho-
coupled aromatic protons indicated an attachment of an aromatic
ring to the 1,4-quinone ring, hence constructing a 1,4-naphtho-
quinone skeleton. The core 1,4-naphthoquinone skeleton of 1 was
confirmed by 13C NMR data for two carbonyl carbons at dC 189.7
(C-1) and 182.9 (C-4), two aromatic methine carbons at dC 120.7
(C-5) and 115.3 (C-6) and six quaternary aromatic carbons at dC
148.2 (C-2), 140.1 (C-3), 124.3 (C-4a), 153.9 (C-7), 151.8 (C-8) and
115.2 (C-8a). The 13C NMR spectrum showed additional signals for
a methoxy carbon at
13.5 (C-13) and the 2-oxopropyl unit at
d
C 56.4 (C-9), an aromatic methyl carbon at dC
C 41.1 (C-10), 203.2 (C-11)
d
sets of ortho-coupled aromatic protons (dH 7.07, d, J = 8.5 Hz and dH
and 29.7 (C-12). Substitution on the naphthoquinone core of 1 was
assigned based on the COSY and HMBC correlations shown in Fig. 2.
The chelated hydroxyl proton was placed at C-8 since it gave HMBC
cross peaks to C-7, C-8 and C-8a. The position of a methoxy group
was assigned at C-7 based on an HMBC correlation of methoxy
protons with C-7. One of the two ortho-coupled aromatic protons at
6.76, d, J = 8.5 Hz, each 2H), indicative of the presence of a 1,4-
disubstituted benzene ring, and four methylene groups (
d
H 4.30, t,
J = 5.0 Hz; 2.87, t, J = 5.0 Hz; 2.67, m; 2.61, m; each 2H). The 13C NMR
spectrum indicated the presence of two carbonyl (dC 175.5 and
172.2),twoaromaticquaternary (dC 129.97and 154.5),twoaromatic
methine (dC 129.99 and 115.4), and four methylene (dC 65.5, 34.1,
29.0 and 28.4) carbons. Methylene protons at dH 2.87 (H2-7)
exhibited 1H–1H COSY correlation with oxymethylene protons atdH
4.30 (H2-8), suggesting a 2-substituted 1-oxyethyl fragment. This
fragment was attached to C-1 (dC 129.97) of the aromatic ring
according to HMBC correlations of H2-7 with C-2, and C-6 and H2-8
with C-1. The other methylene group at dH 2.67 (H2-11) was
connected to the remaining methylene group at dH 2.61 (H2-12)
based on the HMBC cross peaks of H2-11 to C-12 and H2-12 to C-11,
indicative of a 1,2-disubstituted ethyl moiety. The two fragments
were connected via an ester linkage on the basis of HMBC
correlations of H2-8, H2-11, H2-12 to C-10 ester carbonyl (dC
172.2), which was further supported by the chemical shift of C-8 (dC
65.5). H2-12 also showed a HMBC cross peak with the other carbonyl
d
H 7.68 was attributed to H-5 on the basis of its HMBC correlations to
C-4, C-6 and C-7. Thus, the other ortho-coupled aromatic proton
would be located at the 6-position, which was supported by its cross
Table 1
1H (500 MHz) and 13C NMR (125 MHz) data of compounds 1 and 2 in CDCl3.
1
2
Position
dH (J in Hz)
dC
Position
dH (J in Hz)
dC
1
189.7
148.2a
140.1a
182.9
124.3
120.7
115.3
153.9
151.8
1
129.97
129.99
115.4
154.5
34.1
2
2,6
3,5
4
7.07 (d, 8.5)
6.76 (d, 8.5)
3
7.68 (d, 8.4)
7.08 (d, 8.4)
4
4a
5
7
2.87 (t, 5.0)
4.30 (t, 5.0)
carbon (dC 175.5) at C-13. Since there were no other proton signals,
12.41 (s)
8
65.5
the substituents at C-4 (dC 154.5) and C-13 must be hydroxyl groups.
6
10
11
12
13
172.2
28.4
Hence, 2 was characterized as a new succinate ester derivative.
Due to paucity of materials for further biological activity
evaluation, compounds 2, 4 and 5 were prepared via short synthetic
sequencesfromthe sameprecursor, tyrosolderivative8, usinga DCC
coupling as a key protocol for ester formation (Scheme 1). In
addition, we have prepared aspergillol A (23) which is the ortho
analogue of 5, and the meta analogue of 5 (22) via the same strategy
in order to evaluate the structure-activity relationship on the
biological activities of 5.
7
3.99 (s)
3.80 (s)
2.67 (m)
2.61 (m)
8
29.0
8-OH
8a
9
175.5
2.34 (s)
2.13 (s)
115.2
56.4
41.1
203.2
29.7
13.5
10
11
12
13
a
Signals can be interchanged.
Please cite this article in press as: Tadpetch, K., et al., Cytotoxic naphthoquinone and a new succinate ester from the soil fungus Fusarium
solani PSU-RSPG227. Phytochem. Lett. (2014), http://dx.doi.org/10.1016/j.phytol.2014.11.018

G Model
PHYTOL 837 1–5
K. Tadpetch et al. / Phytochemistry Letters xxx (2014) xxx–xxx
3
Fig. 2. 1H–1H COSY and HMBC (H ! C) correlations of 1 and 2.
116
117
118
119
120
121
122
123
124
125
126
127
Compound 1 and synthetic compounds 2, 4, 5, 22 and 23 were
evaluated for biological activities including antimalarial (against
Plasmodium falciparum K1) and antimycobacterial (against Myco-
bacterium tuberculosis H37Ra strain) activities as well as cytotoxi-
cities against oral human carcinoma cells (KB), breast cancer cells
(MCF-7) and noncancerous Vero cells (African green monkey
kidney fibroblasts) (Table 2). All of the compounds tested showed
no antimycobacterial activity against M. tuberculosis. Only
compound 1 displayed weak antimalarial activity with the IC50
was nearly as potent as the standard compound tamoxifen against 128
MCF-7 cells while displayed relatively low cytotoxicity to Vero cell 129
lines. All of the biological activities tested for 1 were comparable to 130
those of javanicin (3) previously reported by Trisuwan et al. (2010). 131
Unfortunately, compounds 2, 4, 5, 22 and 23 were inactive to all 132
assays tested. These results implied that a hydroxyl substituent on 133
the right phenyl ring of 5, 22 and 23 had no effect on cytotoxicity 134
against MCF-7 cell lines. Our findings were consistent with those of 135
Pettit et al. (2009) that aspergillol B (5) was inactive against breast 136
cancer cell lines. However, these results were inconsistent with 137
value of 26.1
mild cytotoxic activity to KB cells (IC50 of 22.6
standard compounds doxorubicin and ellipticine. Interestingly, 1
m
M. For cytotoxicity assays, compound 1 exhibited
m
M) compared to
their report in which aspergillol
A (23) showed moderate 138
cytotoxicity against the breast adenocarcinoma MCF-7 with GI50 139
Scheme 1. Preparation of compounds 2, 4, 5, 22 and 23.
Please cite this article in press as: Tadpetch, K., et al., Cytotoxic naphthoquinone and a new succinate ester from the soil fungus Fusarium
solani PSU-RSPG227. Phytochem. Lett. (2014), http://dx.doi.org/10.1016/j.phytol.2014.11.018

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PHYTOL 837 1–5
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K. Tadpetch et al. / Phytochemistry Letters xxx (2014) xxx–xxx
Table 2
sequences within the group was 95.0–99.7%. The ITS phylogeny
supported the identification of the soil fungus PSU-RSPG227 as F.
solani.
174
175
176
Biological activities of compounds 1–5, 22 and 23.
Compound
Antimalarial
Cytotoxicity, IC50 (mM)
IC50 (mM)
KB cells
MCF-7
cells
Vero
cells
3.3. Fermentation and isolation
177
1
26.1
22.6
21.3
179.3
Inactive
170e
The soil fungus F. solani PSU-RSPG227 were cultured twice. The
fermentations and extractions were performed using the same
procedure described previously by Rukachaisirikul et al. (2007).
The broth EtOAc extracts were obtained as dark brown gums in
1.7 g and 1.1 g, respectively. The first broth EtOAc extract was
fractionated by column chromatography (CC) over Sephadex LH-
20 with 50% MeOH/CH2Cl2 to afford 4 subfractions (1A–1D).
Further purification of subfraction 1C by a series of CC over
Sephadex LH-20 (50% MeOH/CH2Cl2) and reversed phase silica gel
yielded 7 (5.1 mg). Purification of subfraction 1D by CC over
Sephadex LH-20 (50% MeOH/CH2Cl2) and preparative TLC with 2%
MeOH/CH2Cl2 (6 runs) afforded 2 (1.7 mg). Subjection of the
second broth EtOAc extract to CC over Sephadex LH-20 using 50%
MeOH/CH2Cl2 as an eluent yielded 3 subfractions (2A–2C). Further
separation of subfraction 2B by a gradient system of 0.5% MeOH/
CH2Cl2 to 100% MeOH gave 7 subfractions (2B1–2B7). Subfraction
2B2 was chromatographed on silica gel using a gradient of MeOH/
CH2Cl2 and subsequent preparative TLC with 80% CH2Cl2/petro-
leum ether (4 runs) to give 3 (2.2 mg). Subfraction 2B3 was further
separated by CC over silica gel using a gradient of MeOH/CH2Cl2,
followed by preparative TLC using 0.1% MeOH/CH2Cl2 (7 runs) and
1% MeOH/CH2Cl2 (5 runs) to give 1 (1.4 mg). Subjection of
subfraction 2B6 to CC over silica gel using a gradient system of
30% EtOAc/petroleum ether to 80% EtOAc/petroleum ether
afforded 6 (5.2 mg) and 5 subfractions (2B6A–2B6E). Further
purification of subfraction 2B6A by multiple preparative TLC gave 4
(5.2 mg) and 5 (2.2 mg).
178
179
180
181
182
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187
188
189
190
191
192
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195
196
197
198
199
200
201
202
203
204
2
3a
Inactive
12b
Inactive
5.7c
Inactive
13d
4
Inactive
Inactive
Inactive
Inactive
–
Inactive
Inactive
Inactive
Inactive
1.1
Inactive
Inactive
Inactive
Inactive
11.5
Inactive
Inactive
Inactive
Inactive
–
5
22
23
Doxorubicinf
Ellipticinef
Tamoxifenf
Dihydroartemisining
–
4.6
–
8.5
–
–
18.7
–
0.0022
–
–
–
–: not evaluated. For cytotoxicity assays, inactive refers to no inhibition at >50
mg/
mL of a compound tested. For antimalarial assay, inactive refers to no inhibition at
>10
a
m
g/mL of a compound tested.
Previous report by Trisuwan et al.
b
c
d
e
f
IC50 of standard compound dihydroartemisinin = 0.004 mM.
IC50 of standard compounds doxorubicin = 0.33
IC50 of standard compound doxorubicin = 2.18
IC50 of standard compound ellipticine = 4.47 M.
m
M and ellipticine = 1.1
mM.
mM.
m
Standard compounds for cytotoxicity assays.
Standard antimalarial drug.
g
140
141
of 7.2 mg/mL. Further structure modification of these analogues
might be warranted these studies.
142
143
3. Experimental
3.1. General experimental procedures
144
145
146
147
148
149
150
151
152
153
154
155
156
157
Ultraviolet (UV) spectra were measured on a SHIMADZU
UV-160A spectrophotometer. Infrared (IR) spectra were recorded
on a Perkin Elmer 783 FTS165 FT-IR spectrometer. Mass spectra
were obtained on a MAT 95 XL Mass Spectrometer (Thermo
Finnigan). 1H, 13C and 2D NMR spectroscopic data were recorded on
either a 300 or 500 MHz Bruker FTNMR Ultra Shield spectrometer.
Solvents for extraction and chromatography were distilled at their
boilingpoint ranges prior to use. All reactions wereperformed under
argon in oven-dried glassware using analytical grade solvents. All
commercially available reagents were used as received. Thin-layer
chromatography (TLC) and preparative TLC (PTLC) were performed
on silica gel GF256 (Merck). Column chromatography was performed
on silica gel 60 (Merck 230–400 mesh ASTM) or on Sephadex LH-20
or on C-18 reversed phase silica gel.
3.4. Solaninaphthoquinone (1)
205
Orange solid: UV (MeOH) lmax nm (log
e
): 221 (3.57), 268
206
207
208
209
(3.49); IR (thin film) n
max 3152, 1716, 1606 cmꢀ1; 1H and 13C NMR
data see Table 1; HREIMS m/z calcd for C15H14O5 [M+] 274.0841,
found 274.0836.
3.5. 4-(4-Hydroxyphenethoxy)-4-oxobutanoic acid (2)
210
White solid: UV (MeOH) lmax nm (log
e
): 223 (1.84); IR (thin
211
212
213
214
film) nmax 3270, 1721, 1612 cmꢀ1 1H and 13C NMR data see
;
Table 1; HREIMS m/z calcd for C12H14O5 [M+] 238.0841, found
238.0846.
3.6. Syntheses of compounds 2, 4, 5, 22 and 23
215
158
3.2. Fungal material
See supplementary data for detailed syntheses of compounds 2,
4, 5, 22 and 23.
216
217
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
The soil fungus F. solani PSU-RSPG227 was isolated from a soil
sample collected from the Plant Genetic Conservation Project
under the Royal Initiative of Her Royal Highness Princess Maha
Chakri Sirindhorn, Rajjaprabah Dam, Surat Thani Province,
Thailand in 2011. This fungus was deposited as PSU-RSPG227 at
the Department of Microbiology, Faculty of Science, Prince of
Songkla University and the National Center for Genetic Engineer-
ing and Biotechnology (BIOTEC) Culture Collection, Thailand (BCC
number 56863).
The fungal colony was woolly to cottony with white to orange
mycelium. Macroconidia presented 3-5 septate, slightly curved
and stout. Microconidia were large and had 1-2 septate. The ITS
rDNA sequences of F. solani PSU-RSPG227 (Genbank accession no.
KC478531) were clustered with several strains of F. solani, which
was supported by bootstrap values of 100%. The similarity of
3.7. Cytotoxicity assays
218
Cytotoxicity assay against African green monkey kidney
fibroblast (Vero) cells was performed in triplicate employing
the colorimetric method reported by Hunt et al. (1999).
Whereas cytotoxicity assay against MCF-7 and KB cells was
carried out using the resazurin microplate assay (O’Brien et al.,
2000). The standard compound for Vero and KB cell lines was
219
220
221
222
223
224
225
226
227
228
229
ellipticine, which had the IC50 values of 2.10 and 1.12
(8.5 and 4.6 M), respectively. The standard compounds for
MCF-7 cell lines were doxorubicin and tamoxifen, which
exhibited the IC50 values of 6.26 and 6.95 g/mL (11.5 and
18.7 M), respectively.
mg/mL
m
m
m
Please cite this article in press as: Tadpetch, K., et al., Cytotoxic naphthoquinone and a new succinate ester from the soil fungus Fusarium
solani PSU-RSPG227. Phytochem. Lett. (2014), http://dx.doi.org/10.1016/j.phytol.2014.11.018

G Model
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K. Tadpetch et al. / Phytochemistry Letters xxx (2014) xxx–xxx
5
Bra¨se, F., Encinas, A., Keck, J., Nising, C.F., 2009. Chemistry and biology of mycotoxins 262
and related fungal metabolites. Chem. Rev. 109, 3903–3990.
230
3.8. Antimalarial assay
263
Changsen, C., Franzblau, S.G., Palittapongarnpim, P., 2003. Improved green fluores- 264
cent protein reporter gene-based microplate screening for antituberculosis 265
compounds by utilizing an acetamidase promoter. Antimicrob. Agents Che- 266
231
232
233
234
Antimalarial activity was evaluated against the parasite P.
falciparum (K1, multi-drug-resistant strain), using the microcul-
ture radioisotope technique based on the method described by
Desjardins et al. (1979).
mother. 47, 3682–3687.
Cheng, M.-J., Wu, M.-D., Chen, I.-S., Yuan, G.-F., 2008. Secondary metabolites from 268
the mycelia of the fungus Monascus pilosus. Chem. Pharm. Bull. 56, 394–397.
267
269
Desjardins, R.E., Canfield, C.J., Haynes, J.D., Chulay, J.D., 1979. Quantitative assess- 270
ment of antimalarial activity in vitro by a semiautomated microdilution tech- 271
235
3.9. Antimycobacterial assay
nique. Antimicrob. Agents Chemother. 16, 710–718.
272
Hunt, L., Jordan, M., De Jesus, M., Wurm, F.M., 1999. GFP-expressing mammalian 273
cells for fast, sensitive, noninvasive cell growth assessment in a kinetic mode. 274
236
237
238
Antimycobacterial activity was determined against M. tubercu-
losis H37Ra using green fluorescent protein (GFP)-based fluores-
cent detection (Changsen et al., 2003).
Biotechnol. Bioeng. 65, 201–205.
275
Kimura, Y., Shimada, A., Nakajima, H., Hamasaki, T., 1988. Structures of naphtho- 276
quinones produced by the fungus, Fusarium sp., and their biological activity 277
toward pollen germination. Agric. Biol. Chem. 52, 1253–1259.
Medentsev, A.G., Akimenko, V.K., 1998. Naphthoquinone metabolites of the fungi. 279
Phytochemistry 47, 935–959.
Nelson, P.E., Dignani, M.C., Anaissie, E.J., 1994. Taxonomy, biology and clinical 281
aspects of Fusarium species. Clin. Microbiol. Rev. 7, 479–504.
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Acknowledgments
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K. Tadpetch thanks The Thailand Research Fund for the TRF
Senior Research Scholar (Grant No. RTA5480002) to Professor V.
Rukachaisirikul. Faculty of Science, Prince of Songkla University is
gratefully acknowledged for partial support. Additional support is
generously provided by the Development and Promotion of
Science and Technology Talents Project (DPST) for L. Jeanmard
and the Science Achievement Scholarship of Thailand (SAST) for A.
Thiraporn.
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Appendix A. Supplementary data
Shu, R.G., Wang, F.W., Yang, Y.M., Liu, Y.X., Tan, R.X., 2004. Antibacterial and 295
xanthine oxidase inhibitory cerebrosides from Fusarium sp, IFB-121, and endo- 296
phytic fungus in Quercus variabilis. Lipids 39, 667–673.
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Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.phytol.2014.11.018.
Stob, M., Baldwin, R.S., Tuite, J., Andrews, F.N., Gillette, K.G., 1962. Isolation of an 298
anabolic, uterotrophic compound from corn infected with Gibberella zeae. 299
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Please cite this article in press as: Tadpetch, K., et al., Cytotoxic naphthoquinone and a new succinate ester from the soil fungus Fusarium
solani PSU-RSPG227. Phytochem. Lett. (2014), http://dx.doi.org/10.1016/j.phytol.2014.11.018