JBIR-37 and -38, Novel Glycosyl Benzenediols, isolated from the sponge-derived fungus, Acremonium sp. SpF080624G1f01

Article abstract of DOI:10.1271/bbb.90346

In the course of our chemical screening program for new secondary metabolites, we isolated new compounds JBIR-37 (1) and -38 (2) from a culture broth of the marine sponge-derived fungus, Acremonium sp. SpF080624G1f01. The structures of 1 and 2 were determ

Full text of DOI:10.1271/bbb.90346

Biosci. Biotechnol. Biochem., 73 (9), 2138–2140, 2009
Note
JBIR-37 and -38, Novel Glycosyl Benzenediols, Isolated
from the Sponge-Derived Fungus, Acremonium sp. SpF080624G1f01
Miho IZUMIKAWA,1 Shams Tabrez KHAN,1;2 Hisayuki KOMAKI,1;2 Aya NAGAI,1 Shigeki INABA,3
y
y
4;
Motoki TAKAGI,1; and Kazuo SHIN-YA
1Biomedicinal Information Research Center (BIRC), Japan Biological Informatics Consortium (JBIC),
2-42 Aomi, Koto-ku, Tokyo 135-0064, Japan
2NITE Biological Resource Center (NBRC), National Institute of Technology and Evaluation (NITE),
2-5-8 Kazusakamatari, Kisarazu, Chiba 292-0818, Japan
3NITE Biotechnology Development Center (NBDC), Department of Biotechnology, National Institute
of Technology and Evaluation (NITE), 2-5-8 Kazusakamatari, Kisarazu, Chiba 292-0818, Japan
4Biomedicinal Information Research Center (BIRC), National Institute of Advanced Industrial Science
and Technology (AIST), 2-42 Aomi, Koto-ku, Tokyo 135-0064, Japan
Received May 13, 2009; Accepted June 7, 2009; Online Publication, September 7, 2009
[doi:10.1271/bbb.90346]
In the course of our chemical screening program for
new secondary metabolites, we isolated new compounds
JBIR-37 (1) and -38 (2) from a culture broth of
the marine sponge-derived fungus, Acremonium sp.
SpF080624G1f01. The structures of 1 and 2 were
determined to be di- and mono-O-ꢀ-D-glucopyranosy-
loxy-4-(1,1-dimethyl-2-propenyl)benzene on the basis of
extensive NMR and MS spectroscopic data, respectively.
of brown rice (Hitomebore), 30 mg of a Bacto-Yeast
extract (BD Biosciences), 15 mg of sodium tartrate
(Kanto Chemical), 15 mg of K2HPO4 (Wako Pure
Chemical) and 45 ml of H2O at 27 ꢀC for 14 d in static
culture.
The culture was extracted with 80% acetone (100 ml).
The resulting extract was evaporated in vacuo, and the
residual aqueous concentrate was partitioned with
EtOAc (100 ml ꢁ 3) before n-BuOH extraction (100
ml ꢁ 2). The n-BuOH layer was evaporated to dryness.
The dried residue (80.2 mg) was chromatographed by
reversed-phase MPLC (Purif-Pack ODS-100, Moritex)
with an H2O–MeOH linear gradient system (0–100%
MeOH). Compounds 1 (5.4 mg) and 2 (0.5 mg) were
respectively isolated as colorless amorphous solids from
the 0–5% MeOH fractions by preparative reversed-
phase HPLC, using a PEGASIL ODS column (Senshu
Pak, 20 i.d. ꢁ 150 mm) with 20% aqueous MeOH
containing 0.4% formic acid (tR 17.3 min) and 30%
aqueous MeOH containing 0.35% formic acid (tR
21.4 min) at a flow rate of 10 ml/min.
Key words: glycosyl benzenediol; marine sponge;
Demospongiae; Acremonium
Many bioactive substances have been isolated from
marine organisms such as marine microbes, phytoplank-
ton, algae, sponges and tunicates.1) In particular, marine-
derived fungi are emerging as an attractive source of new
bioactive compounds.2) Indeed, cytotoxic metabolites
such as diketopiperazine alkaloids,3) trichodermatides4)
and carbonarones5) have been isolated from marine-
derived fungi. We have also reported the new sesqui-
terpenes, JBIR-27 and -28, from a tunicate-derived
fungus, Penicillium sp. SS080624SCf1,6) and the new
aspochracin derivative, JBIR-15, from a marine sponge-
derived fungus, Aspergillus sclerotiorum Huber
Sp080903f04.7) We therefore attempted to isolate fungi
from the marine sponge, Demospongiae, to obtain novel
substances from the fungal culture broths of marine
origin. We isolated two novel compounds designated as
JBIR-37 (1) and -38 (2) from the culture broth of
Acremonium sp. SpF080624G1f01. This paper describes
the fermentation, isolation and structural elucidation of
1 and 2.
The physico-chemical properties of 1 and 2 are
summarized in Table 1. The molecular formulas of 1
and 2 were respectively established as C23H34O12 and
C17H24O7 on the basis of an HR-ESI-MS analysis.
1
The 13C- and H-NMR spectral data for 1 and 2 are
shown in Table 2. The direct connectivity of these
protons and carbons was established by HSQC spectra.
Since the 1H-NMR signals of 1 assignable to sugar
moieties were highly overlapped, the structure was
elucidated by using 2 which possessed a single sugar
moiety. The structure of 2 was elucidated by a series of
NMR analyses involving DQF-COSY and CT-HMBC8)
spectra. Ortho-coupling between aromatic protons 5-H
and 6-H, which were meta-coupled to 2-H, indicated the
presence of a 1,3,4-trisubstituted benzene ring moiety.
By taking into consideration the low-field 13C chemical
The
producing
fungus,
Acremonium
sp.
SpF080624G1f01, was isolated from the marine sponge,
Demospongiae, collected from Ishigaki Island, Okinawa
Prefecture, Japan. The SpF080624G1f01 strain was
cultured in a 500-ml Erlenmeyer flask containing 15 g
y
To whom correspondence should be addressed. Motoki TAKAGI, Fax: +81-3-3599-8494; E-mail: motoki-takagi@aist.go.jp; Kazuo SHIN-YA,
E-mail: k-shinya@aist.go.jp
Abbreviations: HPLC, high-performance liquid chromatography; MPLC, medium-performance liquid chromatography; HR-ESI-MS, high-
resolution-electrospray ionization-mass spectroscopy; DQF-COSY, double quantum filtered-correlation spectroscopy; HSQC, heteronuclear single
quantum coherence; CT-HMBC, constant time heteronuclear multiple bond connectivity

Novel Glycosyl Benzenediols from a Sponge-Derived Fungus
2139
Table 1. Physico-Chemical Properties of 1 and 2
1
2
Appearance
25
colorless amorphous solid
ꢃ38:1ꢀ (c 0.27, MeOH)
colorless amorphous solid
ꢃ42:0ꢀ (c 0.05, MeOH)
C17H24O7
a
½ꢁꢂ
D
Molecular formula
HR-ESI-MSb (m=z)
C23H34O12
found: 525.1937 ½M þ Naꢂþ
calcd.: 525.1948
found: 363.1396 ½M þ Naꢂþ
calcd.: 363.1420
UVc ꢂmax (MeOH) nm (")
IRd ꢃmax (KBr) cmꢃ1
224 (9,650), 280 (2,680)
1631, 1596 (benzene)
224 (6,010), 280 (2,540)
1635, 1596 (benzene)
aOptical rotation was measured with a Horiba SEPA-300 polarimeter.
bHR-ESI-MS data were recorded on a Waters LCT-Premier XE.
cThe UV spectrum was measured by a Beckman Coulter DU730 UV/Vis spectrophotometer.
dThe IR spectrum was obtained with a Horiba FT-720 spectrophotometer.
Table 2. 13C- and 1H-NMR Data for 1 and 2
1
2
Position
ꢄC
ꢄH (integral, mult, J in Hz)
ꢄC
ꢄH (integral, mult, J in Hz)
1
2
3
4
5
6
7
8
9
153.7
117.8
152.6
138.6
116.1
116.0
41.9
152.1
115.4
150.2
138.3
116.2
114.0
41.5
7.12 (1H, d, 2.5)
6.83 (1H, d, 2.6)
7.10 (1H, d, 9.0)
6.98 (1H, dd, 9.0, 2.5)
7.03 (1H, d, 8.8)
6.67 (1H, dd, 8.8, 2.6)
149.8
110.5
6.34 (1H, dd, 17.6, 10.8)
5.03 (1H, d, 17.6)
5.00 (1H, d, 10.8)
1.53 (3H, s)
149.8
110.3
6.32 (1H, dd, 17.4, 10.7)
5.04 (1H, d, 17.4)
5.01 (1H, d, 10.7)
1.50 (3H, s)
10
11
10
28.0
27.7
101.7
75.1
78.6
71.4
78.2
62.6
28.1
27.5
101.5
74.8
78.0
71.0
77.5
62.3
1.52 (3H, s)
4.94 (1H, m)
1.51 (3H, s)
4.94 (1H, d, 7.8)
20
3.54 (1H, dd, 8.8, 7.9)
3.49 (1H, t, 8.8)
3.44 (1H, m)
3.55 (1H, t, 8.2)
3.52 (1H, t, 8.6)
3.44 (1H, t, 9.0)
30
40
50
3.47 (1H, m)
3.93 (1H, m)
3.73 (1H, m)
3.47 (1H, ddd, 9.0, 5.4, 1.6)
3.95 (1H, dd, 12.0, 1.6)
3.77 (1H, dd, 12.0, 5.4)
60
100
200
300
400
500
600
103.3
75.0
78.0
71.4
78.1
62.5
4.81 (1H, d, 7.3)
3.47 (1H, m)
3.48 (1H, m)
3.41 (1H, m)
3.42 (1H, m)
3.93 (1H, m)
3.74 (1H, m)
The 13C- (150 MHz) and 1H- (600 MHz) NMR spectra were recorded on a Varian NMR System 600 NB CL. The samples were measured in CD3OD, and the solvent
peak was used as an internal standard (ꢄH 3.35 and ꢄC 49.0).
shifts of C-1 and C-3 and the high-field shift of C-2, the
oxygen atoms should be substituted at the C-1 and C-3
positions (Fig. 1, right). Singlet methyl protons 10-H
and 11-H were 1H-13C long-range coupled to each other
and commonly coupled to aliphatic quaternary carbon
C-7 and olefinic methine carbon C-8, which in turn were
1H spin coupled to olefinic exomethylene protons 9-H.
These results established a 3-methyl-butenyl moiety
(Fig. 1, right). The 1H-13C long-range correlations in the
HMBC spectrum from methyl protons 10-H and 11-H to
aromatic carbon C-4, and from aromatic proton 5-H to
quaternary carbon C-7 determined the 3-methyl-butenyl
moiety to be substituted at the C-4 position in the
benzene ring.
protons 20-H, 30-H, 40-H and 50-H. In addition to these
coupling constants, 1H-13C long-range coupling from
10-H to C-50 elucidated a ꢀ-glucoside moiety (Fig. 1,
right). The direct 13C-1H coupling constant at C-10
(1JC–H ¼ 156:5 Hz)9) also supported the ꢀ-glucopyrano-
side moiety. The location of this glucopyranoside was
1
determined by H-13C long-range coupling from 10-H
to C-3. Therefore, the structure of 2 was determined as
1-hydroxy-3-O-ꢀ-glucopyranosyloxy-4-(1,1-dimethyl-2-
propenyl)benzene (Fig. 1, right).
The structure of 1 was also deduced to be 1,3-di-O-ꢀ-
glucopyranosyloxy-4-(1,1-dimethyl-2-propenyl)benzene
after confirming the aglycon of 1 to be the same as that
of 2 by analyses of the DQF-COSY and CT-HMBC
spectra. Although it was not possible to analyze the
coupling constants among proton signals due to the high
degree of overlapping, two sugar moieties were recog-
The remaining unit was deduced to be a sugar residue
by establishing the sequence from anomeric proton 10-H
to hydroxymethyl protons 60-H through oxymethine

2140
M. IZUMIKAWA et al.
OH
6'
OH
6'
OH
4'
4'
O
O
5'
9
O
9
HO
8
7
HO
O
6
2'
5'
1'
8
HO
HO
O
2'
3'
3
10
OH
2
3'
1'
OH
6''
7
10
3
4
4
5
2
11
1
4''
5''
11
1
O
1''
HO
5
HO
HO
2''
6
3''
OH
JBIR-37 (1)
JBIR-38 (2)
Fig. 1. Structures of 1 and 2, and Key Correlations of 1H-1H (bold line) and 1H-13C (arrows) of 2.
nized in 1. We thus carried out an acid hydrolysis of 1
to determine the absolute configuration of these sugar
moieties. As a result, we obtained only a glucopyrano-
side as a sugar moiety (ꢁ-form, 1-H (ꢄH 5.02, t,
J ¼ 4:0 Hz), 2-H (ꢄH 3.32, dd, J ¼ 9:0, 4.0 Hz), 3-H
(ꢄH 3.50, t, J ¼ 9:0 Hz), 4-H (ꢄH 3.20, t, J ¼ 9:0 Hz),
5-H (ꢄH 3.62, m), 6-H (ꢄH 3.63, dd, J ¼ 12:0, 3.0 Hz, ꢄH
3.55, dd, J ¼ 12:0, 6.0 Hz); ꢀ-form, 1-H (ꢄH 4.44, d,
J ¼ 8:0 Hz), 2-H (ꢄH 3.04, t, J ¼ 8:5 Hz), 3-H (ꢄH 3.28,
t, J ¼ 9:0 Hz), 4-H (ꢄH 3.18, t, J ¼ 9:0 Hz), 5-H (ꢄH
3.25, ddd, J ¼ 9:0, 6.0, 2.0 Hz) and 6-H (ꢄH 3.69, dd,
Acknowledgments
This work was supported by a grant from the New
Energy and Industrial Technology Department Organ-
ization (NEDO) of Japan. Authors thank Mr. Akihiko
Kanamoto of Op Bio Factory Co. Ltd. for his help in
collecting the sponge sample.
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