A new dipeptide isolated from the bulb of garlic

Article abstract of DOI:10.1080/10286020.2013.868886

(R)-3-(allylthio)-2-((R)-3-(allylthio)-2-aminopropanamido)propanoic acid was isolated from the bulb of garlic, together with four known amino acids. Its structure was elucidated on the basis of 2D NMR and MS techniques. To the best of our knowledge, (R)-3-(allylthio)-2-((R)-3-(allylthio)-2-aminopropanamido) propanoic acid, which showed antibacterial activity against the Staphylococcus aureus antibiotic resistant strain, was the first example of dipeptide from garlic.

Full text of DOI:10.1080/10286020.2013.868886

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A new dipeptide isolated from the bulb
of garlic
ab
a
b
b
Mei-Yun Zhou , Zhao-Jun Wu , Wen-Qing Li , Xing-Peng Hu , Rian
b
b
b
Yan , Shi-Yi Ou & Hua Zhou
a
Department of Chemistry, Jinan University, Guangzhou, 510632,
P.R. China
b
Department of Food Science and Engineering, Jinan University,
Guangzhou, 510632, P.R. China
Published online: 16 Dec 2013.
To cite this article: Mei-Yun Zhou, Zhao-Jun Wu, Wen-Qing Li, Xing-Peng Hu, Rian Yan, Shi-Yi Ou &
Hua Zhou (2014) A new dipeptide isolated from the bulb of garlic, Journal of Asian Natural Products
Research, 16:3, 323-326, DOI: 10.1080/10286020.2013.868886
To link to this article: http://dx.doi.org/10.1080/10286020.2013.868886
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Journal of Asian Natural Products Research, 2014
Vol. 16, No. 3, 323–326, http://dx.doi.org/10.1080/10286020.2013.868886
A new dipeptide isolated from the bulb of garlic
a,b
Mei-Yun Zhou , Zhao-Jun Wu , Wen-Qing Li , Xing-Peng Hu , Rian Yan , Shi-Yi Ou
a
b
b
b
b
b
and Hua Zhou *
a
b
Department of Chemistry, Jinan University, Guangzhou 510632, P.R. China; Department of Food
Science and Engineering, Jinan University, Guangzhou 510632, P.R. China
(
Received 11 May 2013; final version received 20 November 2013)
(
R)-3-(allylthio)-2-((R)-3-(allylthio)-2-aminopropanamido)propanoic acid was iso-
lated from the bulb of garlic, together with four known amino acids. Its structure
was elucidated on the basis of 2D NMR and MS techniques. To the best of our
knowledge, (R)-3-(allylthio)-2-((R)-3-(allylthio)-2-aminopropanamido)propanoic
acid, which showed antibacterial activity against the Staphylococcus aureus antibiotic
resistant strain, was the first example of dipeptide from garlic.
Keywords: dipepeptide; garlic; antibacterial activity
1
.
Introduction
oil-macerated garlic [8,9]. As a part of our
continuing studies on secondary metab-
olites from a medical plant or food, we
isolated one new dipeptide 3 from newly
unearthed garlic (Figure 1), together with
four known amino acids L-deoxyalliin 1
Garlic (Allium sativum L.) is used world-
wide as a spice, food, and folk medicine
[1–3]. Many medicinal substances derived
from garlic had been reported so far, most
of which are organosulfur compounds [4–
[13,14], alliin 2 [15], tryptophan, and
phenylalanine.
8
]. The types of compounds isolated from
garlic are intimately related to the methods
of processing garlic [9–12], because alliin,
the major sulfur-containing amino acid in
garlic, is easily converted to diallyl
thiosulfinate (allicin) via alliinase action
when the raw garlic bulb is chopped or
crushed. Furthermore, allicin, as the above
reaction product, is considerably unstable
and easily decomposed into other kinds of
organosulfur compounds [10]. It is natural
that different processing technologies of
garlic could result in different medical
effects. It is said that eating newly
unearthed garlic, rather than any others,
is good for our heath. However, to the best
of our knowledge, the chemical investi-
gation on the newly unearthed garlic has
not been carried out till now, though there
are many studies on the chemical com-
ponents and bioactivities of garlic such as
2
.
Results and discussion
(R)-3-(allylthio)-2-((R)-3-(allylthio)-2-
aminopropanamido)propanoic acid,
obtained as an amorphous, optically
active powder, had the molecular formula
C H N O S , as established by HR-ESI-
1
2 20 2 3 2
þ
MS analysis (m/z 305.0952 [M þ H] )
1
3
and confirmed by C NMR spectroscopy.
The IR spectrum of 3 showed absorption
bands for carbonyl (1686, 1753) and CvC
1
3
groups (1612). The C NMR spectrum
(Table 1) showed a total of 12 signals (4
CH, 6 CH , and 2 quaternary C atoms), in
2
agreement with the molecular formula.
1
13
The H and C NMR spectra were rather
similar to those of 1 [16], one of the major
sulfur-containing amino acids present in
*Corresponding author. Email: zhouhua5460@jnu.edu.cn
q 2013 Taylor & Francis

3
24
M.-Y. Zhou et al.
H N
7' COOH
O
S
NH2
2
NH2
O
2'
6
6'
S
4
S
4'
S
N
COOH
2
COOH
3
5 H 7
H
H
1
3'
H
H
1'
1
2
3
Figure 1. The structures of compounds 1–3.
the garlic, but the number of carbon
3-(allylthio)-2-aminopropanamido)propa-
noic acid, was established. To the best of
our knowledge, 3 was the first example of
dipeptide isolated and elucidated from
garlic.
1
3
signals of 3 in the C NMR spectrum was
two times as those of 1, which suggested
that 3 might be a new dimeric of 1. The
structure of 3 was further revealed by
detailed analysis of one- and two-dimen-
Compounds 1–3 showed inhibitory
activities against Staphylococcus aureus
(compound 1, 370 mg/disk, zone diameter:
8 mm; compound 2, 350 mg/disk, zone
diameter: 12 mm; compound 3, 90 mg/
disk, zone diameter 9 mm).
1
1
sional NMR spectra, as follows. H– H
COSY and HMQC experiments indicated
the presence of the following structures:
CH vCHCH and CH CH (NH )C(O),
2
2
2
2
which was further confirmed by a HMBC
experiment (Figure 2). The above-men-
tioned two fragments were linked by an S
atom based on the HMBC cross-peaks H-
3.
Experimental
3
.1 General experimental procedures
5
C(6 ) on the basis of the HMBC cross-
/C-3. A signal at d 53.7 was assigned to
0
C
Optical rotations were measured with a
Jasco DIP-370 digital polarimeter (Tokyo,
Japan). IR spectra were obtained on an IR-
450 infrared spectrophotometer (Tokyo,
Japan) with KBr pellets. FAB-MS and HR-
FAB-MS were carried out on an Autospe-
3000 spectrometer (Manchester, UK).
Nuclear magnetic resonance spectra were
recorded on Bruker AM 500 MHz spec-
trometer (Bruker, Fallanden, Switzerland).
0
0
0
peaks H-6 /C-7, C-7 , and C-5 .
The configuration of 3 was revealed
through acid hydrolysis of 3. The obtained
product of acid hydrolysis was found to be
identical to 1 because it has the same
25
optical rotation (½aꢀ 213.1 (c ¼ 2,
D
1
H O)) and H NMR spectrum as 1.
2
Therefore, the absolute configurations of
0
3
at C(6) and C(6 ) were both elucidated to
be (R).
On the basis of the above discussion
d in ppm rel. to Me Si as internal standard.
4
Solvents were distilled before use. TLC
and columm chromatography (CC) were
carried out on plates precoated with gel
and by comparison with the literature data
of 1 [16], the structure of the new
dipeptide, named (R)-3-(allylthio)-2-((R)-
F254 and silica gel H (SiO , Qingdao
2
1
Table 1. H and C NMR spectral data of compound 3 in MeOD (500 MHz for H and 125 MHz
13
1
1
for C, res.; d in ppm).
3
Position
d
C
d
H
Position
d
C
d
H
0
1
2
3
5
119.0
134.5
35.0
33.1
53.0
5.24–5.28 (m)
5.83–5.91 (m)
3.26–3.26 (m)
2.99–3.04 (m)
4.24–4.27 (m)
–
1
2
3
5
6
7
118.3
135.2
35.7
32.5
53.7
5.09–5.13 (m)
5.70–5.76 (m)
3.10–3.18 (m)
2.85–2.90 (m)
4.60–4.65 (m)
–
0
0
0
0
6
0
NH2
7
CONH
168.1
173.2
8.81
4.60

Journal of Asian Natural Products Research
325
2
(cm ): 3002, 2633, 1753, 1686, 1612, 87,
1
NH2
COOH
1 13
440, 783. For H and C NMR spectral
O
1
data, see Table 1. ESI-MS: m/z 305 (100)
S
S
N
H
þ
þ
[
M þ H] . HR-ESI-MS: m/z 305.0952
[
M þ H] (calcd for C H N O S ,
1
2
21
2
3 2
3
05.0988).
HMBC
COSY
Figure 2. Key HMBCs and COSYs for 3.
3.4 Acid hydrolysis of 3
During the study, 1.0 ml of a 3.0 g/l
aqueous solution of 3 was treated with
Haiyang Chemical Co. Ltd, Qingdao,
China).
10.0 mol/l HCl (1.0 ml) at 808C for 18 h.
The mixture was immediately purified by
acid cation exchange resin (Amberlitew
IRP-69, Shanghai Aladdin Industrial Cor-
poration, Shanghai, China) by using
3
.2 Plant material
The bulb of garlic was collected in Boluo
city, Guangdong Province of China and
identified by professor Shi-Yi Ou (Jinan
University). A specimen of the plant has
been deposited in the Department of Food
Science and Engineering, Jinan Univer-
sity, Guangzhou, China.
NH OH (30%) to afford 1 (2.0 mg),
4
1
which was identified by H NMR and
TLC and optical rotations measurement.
3.5 Antibacterial assay
S. aureus was spread with an aseptic
spreader on the surface of the agar medium
3
.3 Extraction and isolation
The garlic (8.0 kg) was frozen by liquid
nitrogen of 2 1968C, and then broken,
extracted with 90% ethanol. After the
removal of the solvent by evaporation, the
residue was dissolved in water and chro-
dish. The samples were dissolved in CH OH
3
and added to each paper disk with syringes,
respectively. The disks were dried with flow
air and put onto agar media inoculated with
the testing organism for 10 h at 428C. The
antibacterial activity was calculated by the
diameter of the inhibition zone.
matographed ona D-101 resin withH O and
2
ethanol to give two fractions. The H O
2
eluent as the main amino acid was
concentrated to obtain 9.0 g crude amino
acid, which was chromatographed on ion-
cation exchange resin (Amberlite IR-120)
with 5% NaOH solution to give four
fractions. Fraction 2 was subjected to CC
Acknowledgements
This work was financially supported by the
NSFC of China (Nos 31000816, 31101323) and
by the high-performance computing platform of
Jinan University.
(
(
silica gel; H O/MeOH; 25:75) to furnish 2
2
150mg), 1 (50 mg), tryptophan (20 mg),
and phenylalanine (15 mg); fraction 4 was
chromatographed on CC (silica gel; pet-
roleumether/AcOEt;2:3)toyield 3(15 mg).
References
[
1] E. Ayaz and H.C. Alpsoy, Turk. Parazitol.
Derg. 31, 145 (2007).
[
[
2] M. Keusgen, Planta Med. 63, 93 (1997).
3] R. Kubec, R.B. Cody, and A.J. Dane, J.
Agric. Food Chem. 58, 1121 (2010).
3
.3.1 (R)-3-(Allylthio)-2-((R)-3-
allylthio)-2-aminopropanamido)
(
propanoic acid
[4] Q.H. Hu, Q. Yang, O. Yamato, M.
Yamasaki, Y. Maede, and T. Yoshihara,
J. Agric. Food Chem. 50, 1059 (2002).
[5] M. Nishida, T. Hada, K. Kuramochi, H.
Yoshida, H. Yonezawa, I. Kuriyama, F.
1
D
9:2
Colorless amorphous powder. ½aꢀ
2
10.1 (c ¼ 0.1, CH OH). IR (KBr) n
3
max

3
26
M.-Y. Zhou et al.
Sugawara, H. Yoshida, and Y. Mizushina,
Food Chem. 108, 551 (2008).
6] M. Thomson and M. Ali, Curr. Cancer
Jeong, Food Sci. Biotechnol. 16, 963
(2007).
11] H. Yoshida, H. Katsuzaki, R. Ohta, K.
Ishikawa, H. Fukuda, T. Fujino, and A.
Suzuki, Biosci. Biotechnol. Biochem. 63,
[
[
Drug Targets 3, 67 (2003).
7] M.J. Wargovich, Carcinogenesis 8, 487
[
5
91 (1999).
(
1987).
[
12] H. Nishimura, T. Takahashi, C.H. Wijaya,
and T. Ariga, Biofactor 13, 257 (2000).
13] A. Stoll, Helv. Chim. Acta 34, 481 (1951).
14] A. Stoll, Helv. Chim. Acta 31, 189 (1948).
15] M.M. Eckner, C.A. Erdelmeier, O.
Sticher, and H.D. Reuter, J. Nat. Prod.
[
8] O. Yamato, Y. Sugiyama, H. Matsuura,
K.W. Lee, K. Goto, M.A. Hossain, Y.
Maede, and T. Yoshihara, Biosci. Bio-
technol. Biochem. 67, 1594 (2003).
[
[
[
[
9] S.J. Ziegle and O. Sticher, Planta Med.
55, 372 (1989).
5
6, 864 (1993).
[
10] I.G. Hwang, K.S. Woo, K.S. Kim, J.T.
Hong, B.Y. Hwang, Y.R. Lee, and H.S.
[16] F. Freeman and B.G. Huang, J. Org.
Chem. 59, 3277 (1994).