Two new compounds from the metabolites of a marine-derived actinomycete Streptomyces cavourensis YY01-17

Article abstract of DOI:10.1080/10286020.2012.762764

Our current marine natural product program investigated the second metabolites of an actinomycete Streptomyces cavourensis YY01-17 originating from the Antarctic ecological niche to discover potential antitumor chemical entities. Two new compounds, along with a known compound, were isolated from the ethyl acetate extract of the fermentation broth of the marine-derived actinomycete, and their structures were elucidated, respectively, as 2(S)-3′-hydroxybutan-2′-yl 2-hydroxypropanoate (1), (E)-3-hydroxy-2,4-dimethylhept-4-enamide (2), and 2-hydroxy-3-methylbutanoic acid (3) on the basis of spectroscopic data interpretation.

Full text of DOI:10.1080/10286020.2012.762764

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Two new compounds from the
metabolites of a marine-derived
actinomycete Streptomyces cavourensis
YY01-17
Shan-Shan Su a , Li Tian b c , Gang Chen a , Zhan-Qiang Li a , Wen-
Feng Xu a & Yue-Hu Pei a
a School of Traditional Chinese Materia Medica, Shenyang
Pharmaceutical University , Shenyang , 110016 , China
b The First Institute of Oceanography SOA , Qingdao , 266061 ,
China
c Qingdao University of Science & Technology , Qingdao , 266042 ,
China
Published online: 19 Feb 2013.
To cite this article: Shan-Shan Su , Li Tian , Gang Chen , Zhan-Qiang Li , Wen-Feng Xu & Yue-Hu Pei
(2013): Two new compounds from the metabolites of a marine-derived actinomycete Streptomyces
cavourensis YY01-17, Journal of Asian Natural Products Research, 15:3, 265-269
To link to this article: http://dx.doi.org/10.1080/10286020.2012.762764
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Journal of Asian Natural Products Research, 2013
Vol. 15, No. 3, 265–269, http://dx.doi.org/10.1080/10286020.2012.762764
Two new compounds from the metabolites of a marine-derived
actinomycete Streptomyces cavourensis YY01-17
Shan-Shan Sua, Li Tianbc, Gang Chena, Zhan-Qiang Lia, Wen-Feng Xua and Yue-Hu Peia*
aSchool of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang
b
c
110016, China; The First Institute of Oceanography SOA, Qingdao 266061, China; Qingdao
University of Science & Technology, Qingdao 266042, China
(Received 6 June 2012; final version received 25 December 2012)
Our current marine natural product program investigated the second metabolites of an
actinomycete Streptomyces cavourensis YY01-17 originating from the Antarctic
ecological niche to discover potential antitumor chemical entities. Two new
compounds, along with a known compound, were isolated from the ethyl acetate
extract of the fermentation broth of the marine-derived actinomycete, and their
structures were elucidated, respectively, as 2(S)-30-hydroxybutan-20-yl 2-hydroxy-
propanoate (1), (E)-3-hydroxy-2,4-dimethylhept-4-enamide (2), and 2-hydroxy-
3-methylbutanoic acid (3) on the basis of spectroscopic data interpretation.
Keywords: marine actinomycete; Streptomyces cavourensis; amide
1. Introduction
2. Results and discussion
Marinemicro-organismscontinuetobeboth
a major focus of many research efforts and a
rich source of novel metabolites all over the
world [1]. Recent years, actinomycetes
isolated from the marine environment
have attracted considerable attention.
Extensive exploration of the second
metabolites of marine actinomycetes has
led to the isolation of many important drug
leads, and some are developed into anti-
microbial, anticancer, and immunosuppres-
sive drugs [2].
Compound 1 was obtained as a colorless oil
with ½a2D0ꢀ þ 6.0 (c 0.05, MeOH), for which
HR-ESI-MS established its molecular
formula as C7H14O4 at m/z 163.0972
[M þ H]þ. Its UV spectrum showed
absorption maximum at 219 nm; and the
IR spectrum revealed absorption bands at
3393, 2982, 2939, 1733, 1452, and
1377 cm21. Analysis of the NMR spectra
(Table 1) revealed chemical shifts indica-
tive of a carboxyl moiety (dC 174.2), two
hydroxyl groups (dH 4.73 and dH 5.27),
three methyl groups [(dH 1.02, dC 19.1), (dH
1.12, dC 15.5), and (dH 1.23, dC 20.5)], and
three oxygenated methine moieties [(dH
3.57, dC 68.0), (dH 4.08, dC 66.2), and (dH
4.60, dC 74.2)]. The presence of these
groups was supported by HSQC exper-
iments. On the basis of the HMBC
correlations (Figure 2) of H-3 (dH 1.23)
with C-2 (dC 66.2); H-10 (dH 1.12) with C-20
(dC 74.2), C-30 (dC 68.0); H-4 (dH 1.02)
with C-30 (dC 68.0), C-20 (dC 74.2), and the
As part of our ongoing screening for
new antitumor compounds from marine
micro-organisms [3], we have isolated a
marineactinomycete Streptomyces cavour-
ensis YY01-17 from the lichens, grown
in the Antarctic area. Studies on the
ethyl acetate extract of the fermentation
broth of the actinomycete afforded three
compounds, among which compounds 1
and 2 (Figure 1) were new linear
compounds.
*Corresponding author. Email: peiyueh@vip.163.com
q 2013 Taylor & Francis

266
S.-S. Su et al.
O
1
O
1'
2'
6
1
NH2
3
2
HO
OH
2
4
O
HO
7
5
3'
OH
4'
3
O
OH
1
2
3
Figure 1. The structures of compounds 1–3.
correlations of OH-2 (dH 5.27) with C-2
(dC 66.2), C-3 (dC 20.5); OH-30 (dH 4.73)
with C-20 (dC 74.2), C-40 (dC 19.1), C-30 (dC
68.0), the complete carbon connectivity
was established (Figure 1). On alkaline
hydrolysis of 1, the optical rotation of lactic
acid was ½a2D0ꢀ 213.8 (c 0.3, H2O) and
L-lactic acid was confirmed by comparing
with the data in the literature [4], so the
absolute configuration of C-2 was con-
firmed as S. Thus, compound 1 was finally
elucidated as 2(S)-30-hydroxybutan-20-yl 2-
hydroxypropanoate.
J ¼ 6.9 Hz)/dC 128.5, dC 135.5], an amide
carbonyl moiety [dH 7.17 (1H, s, ZNH),
6.65 (1H, s, ZNH), and dC 177.2], an
ethyl group [dH 0.91 (3H, t, J ¼ 7.5 Hz)/dC
14.2, dH 1.96 (2H, m)/dC 20.3], an
oxygenated alcohol methine group [dH
3.81 (1H, d, J ¼ 9.3 Hz)/dC 78.9, dH 4.68
(1H, br s, ZOH)], a methyl moiety adjacent
to an unsaturated quaternary carbon [dH
1.49 (3H, s)/dC 10.6], and a tertiary
aliphatic methyl group [dH 0.74 (3H, d,
J ¼ 7.2 Hz)/dC 15.0, dH 2.27 (1H, m)/dC
43.1]. The complete connectivity of these
groups was confirmed by HMBC corre-
lations (Figure 2) of H-5 at dH 5.24 with 4-
CH3 at dC 10.6, C-7 at dC 14.2, C-6 at dC
20.3, C-3 at dC 78.9; 4-CH3 at dH 1.49 with
C-3 at dC 78.9, C-5 at dC 128.5, C-4 at dC
135.5; and H-3 at dH 3.81 with 4-CH3 at dC
10.6, 2-CH3 at dC 15.0, C-2 at dC 43.1, C-5
at dC 128.5, C-4 at dC 135.5, C-1 at dC
177.2. The absence of any correlation
between CH3-4 and H-5 in the NOESY
spectrum (Figure 3), meanwhile the
obvious cross-peak between CH3-4 and
Compound 2 was also obtained as a
colorless oil with ½a2D0ꢀ þ 12.0 (c 0.05,
MeOH), and its molecular formula was
established to be C9H17NO2 by HR-ESI-
MS at m/z 172.1334 [M þ H]þ, thereby
indicating two degrees of unsaturation. Its
UV spectrum showed absorption maximum
at 210 and 238 nm; and the IR spectrum
revealed absorption bands at 3339, 2966,
2934, 1662, 1609, 1459, and 862 cm21. The
NMR spectra (Table 1) of 2 illustrated a
trisubstituted double bond [dH 5.24 (1H, t,
Table 1. 1H NMR (300 MHz) and 13C NMR (75 MHz) spectral data of compounds 1 and 2 in
DMSO-d6.
1
2
Position
dC
d
H (J in Hz)
Position
dC
dH (J in Hz)
1
2
174.2
66.2
20.5
15.5
74.2
68.0
19.1
–
–
1
2
3
4
5
177.2
43.1
78.9
135.5
128.5
20.3
14.2
15.0
10.6
–
–
2.27 (1H, m)
3.81 (1H, d, 9.3)
–
5.24 (1H, t, 6.9)
1.96 (2H, m)
0.91 (3H, t, 7.5)
0.74 (3H, d, 7.2)
1.49 (3H, s)
4.08 (1H, m)
1.23 (3H, d, 6.6)
1.12 (3H, d, 6.0)
4.60 (1H, m)
3
10
20
30
3.57 (1H, m)
6
7
40
1.02 (3H, d, 6.3)
4.73 (1OH, d, 5.1)
5.27 (1OH, d, 5.7)
30-OH
2-OH
2-CH3
4-CH3
3-OH
ZNH2
–
4.68 (1H, br s)
7.17 (1H, s), 6.65 (1H, s)
–

Journal of Asian Natural Products Research
267
O
CH3
CH
CH3
CH3
CH
H2
C
HO
NH2
OH
H
C
H
H
C
C
O
H3C
C
H
CH3
OH
CH3
O
1
2
Figure 2. The key HMBC correlations of new compounds 1 and 2.
H-6, supported the (E)-double-bond geo-
metry. Hence, the structure of compound 2
was finally determined as (E)-3-hydroxy-
2,4-dimethylhept-4-enamide. Meanwhile,
the relative configuration of this compound
was confirmed by the NOESY spectrum in
Newman projection (Figure 3). The
appearance correlations of H-3 with H-2
and CH3-2 declared H-3 was in place
between H-2 and CH3-2 in Newman
projection. The obvious cross-peak
between CH3-2 and CH3-4 supported that
CH3-4 was surrounding CH3-2. So the
elative configuration of compound 2 was
confirmed as shown in Figure 3.
were measured on a Perkin-Elmer 241
polarimeter (Perkin-Elmer Co., Jena,
Germany). The NMR spectral data were
recorded on Bruker AV-600 (600 MHz for
1H and 150 MHz for 13C) with TMS
(tetramethylsilane) as the internal standard
(Bruker Co.). The HR-ESI-MS data were
obtained on the Microssmass AutoSpec-
UltimaE TOF mass spectrophotometer
(Bruker Co.). Chromatography was carried
out on silica gel (200–300 mesh; Qingdao
Haiyang Chemical Factory, Qingdao,
China), Sephadex LH-20 (Pharmacia,
Piscataway, NJ, USA), and reversed-
phase HPLC (Shimadzu LC-8A vp,
Kyoto, Japan).
The known compound 2-hydroxy-3-
methylbutanoic acid (3) was identified by
the comparison of the spectral data (1H and
13C NMR) with those reported in the
literature [5].
3.2 Actinomycete material
The actinomycete was isolated from the
lichens, collected from Antarctica in
December 1999. A voucher specimen
(No. HTTA-F99123), the genomic DNA
of strain, was prepared, and 16S rDNA
genes were amplified with a method
described by Kataoka et al. [6].
3. Experimental
3.1 General experimental procedures
UV spectra were measured on a Shimadzu
UV-1601 (Shimadzu, Tokyo, Japan). IR
spectra were measured on a Bruker IFS-55
infrared spectrophotometer (Bruker Co.,
Zurich, Switzerland). Optical rotations
The 16S rDNA sequences were aligned
with published sequences from the Gene-
Bank database (http://www.ncbi.nlm.nih.
gov) using the NCBI BLASTN comparison
software. Phylogenetic trees were con-
structed by the neighbor-joining method
using the DNAMAN software (version 5.1,
Lynnon Biosoft, Quebec, Canada) (Figure
4). The closest relatives of the remaining
sequences were obtained from the Gene-
Bank database using BLAST program and
sequence similarity was 99% between
strain YY01-17 and S. cavourensis
NR04385.1. From phylogenetic analysis,
physiological and biochemical tests, and
CH3
CH3
H
CONH2
H
OH
Figure 3. The key NOESY correlations of
new compound 2.

268
S.-S. Su et al.
Figure 4. Phylogenetic tree of species YY01-17 based on 16S rDNA gene sequences.
morphological characteristics, the strain
was identified as S. cavourensis (GeneBank
No. GU575124), and has been deposited in
the Marine Microbial Medicinal Resource
Library of the First Institute of Oceanogra-
phy SOA funded by the Ministry of Science
and Technology.
Sephadex LH-20 column chromatography
(CH3OH) and preparative HPLC (CH3-
OHZH2O 58.5:100, flow rate 1.0 ml/min,
wavelength 210 nm) to obtain 2 (7.7 mg,
retention time 34 min).
3.3.1 2(S)-30-Hydroxybutan-20-yl 2-
hydroxypropanoate (1)
Colorless oil, ½a2D0ꢀ þ 6.0 (c 0.05, MeOH);
UV (MeOH) lmax: 219 nm; IR (KBr) nmax
(cm21): 3393, 2982, 2939, 1733, 1452,
1377; 1H NMR and 13C NMR spectral
data, see Table 1; HR-FAB-MS m/z:
163.0972 [M þ H]þ (calcd for C7H15O4,
163.0970).
3.3 Fermentation, extraction, and
isolation
The inoculum prepared by putting Petri
dish cultures of YY 01-17 into 500-ml
baffled flasks containing 150 ml of the
broth (soluble starch 1.8%, peptone 0.2%,
seawater 25% in deionized water, pH 7.0)
was fermented by shaking at 150 rpm and
208C for 20 days in incubator shaker.
The supernatant of the fermentation
broth of the strain YY 01-17 (50 liters) was
concentrated to 5 liters in vacuo and
extracted with ethyl acetate and n-butanol,
successively. The EtOAc crude extract
(10.7 g) was subjected to silica gel column
and eluted with CHCl3ZCH3OH (100:1–
0:1), yielding eight fractions. Fraction 3
(1.2 g) was purified by Sephadex LH-20
column chromatography (CH3OH) and
preparative HPLC (CH3OH–H2O 40:100,
flow rate 1 ml/min, wavelength 210 nm) to
obtain compounds 1 (15.8 mg, retention
time 25 min) and 3 (15 mg, retention time
32 min). Fraction 4 (2.3 g) was purified by
3.3.2 (E)-3-Hydroxy-2,4-dimethylhept-
4-enamide (2)
Colorless oil, ½a2D0ꢀ þ 12.0 (c 0.05,
MeOH); UV (MeOH) lmax: 210, 238 nm;
IR (KBr) nmax (cm21): 3339, 2966, 2934,
1
1662, 1609, 1459, 862; H NMR and 13C
NMR spectral data, see Table 1; HR-FAB-
MS m/z: 172.1334 [M þ H]þ (calcd for
C9H18NO2, 172.1338). This compound
could get CAS Registry Number as
1246085-99-3 but without any reference.
3.3.3 Alkaline hydrolysis of compound 1
Compound 1 was hydrolyzed with 5%
KOH in dioxane (1 ml) for 5 h. The

Journal of Asian Natural Products Research
269
AE06B04), and the National Natural Science
Foundation of China (40976104).
mixture was extracted with n-butanol, and
then aqueous fraction was neutralized with
5% HCl. The aqueous fraction was
detected by optical rotation analysis and
L-lactic acid was confirmed by comparing
with the data in the literature.
References
[1] J.W. Blunt, B.R. Copp, R.A. Keyzers,
M.H.G. Munro, and M.R. Prinsep, Nat.
Prod. Rep. 29, 144 (2012).
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Acknowledgements
This work was financially supported by the
National High Technology Research and
Development Program (863 Program) of
China (2011AA10A202-2), National Key
Technologies R&D Program (2011B
[6] M. Kataoka, K. Ueda, and T. Kudo,
Microbiol. FEMS Lett. 151, 249 (1997).