Catechoserine from Streptomyces sp.
Yasuhiro Igarashi et al
209
Isolation
Marfey’s analysis
At the end of the fermentation period, the whole culture broth (2l) was A portion of 1 (0.5mg) was hydrolyzed at 1101C with 6 N HCl (200ml) for
centrifuged at 6000r.p.m. for 10min, and the supernatant was separated. The 15h, and the reaction mixture was evaporated to dryness. A 0.1 M NaHCO3
mycelium was extracted with 1 l of MeOH at room temperature for 3 h, and the solution (100ml) was added to the dried hydrolysate of 1, as well as to standards
solution was recovered by filtration. After repeating this extraction process of L-Ser and D-Ser. A solution of L-FDLA in acetone (0.05 mg in 50ml) was
twice, the combined organic solution was concentrated under reduced pressure. added to each reaction tube. Each tube was sealed and incubated at 501C for
The remaining aqueous solution was combined with the supernatant and 30min. To quench reactions, 50 ml of 2 N HCl was added and then diluted with
charged onto a column of HP-20 resin, which was eluted with a step gradient of 100 ml of MeOH/0.2% HCO2H (50 : 50). The Marfey’s derivatives of the
MeOH/distilled water (increasing MeOH by 20% from 0 to 100% MeOH). hydrolysate and standards were analyzed by HPLC using a Cosmosil 5C18-
Compounds 1, 2 and 3 were detected in fractions eluted with 100%, 60% and AR-II column (Nacalai Tesque, Kyoto, Japan, 4.6ꢁ250 mm) eluted with MeCN-
80% MeOH, respectively, by HPLC. The 100% MeOH fraction was concen- 2% HCO2H in H2O at a flow rate of 1.0 mlminꢀ1, monitoring at 340 nm. The
trated under reduced pressure to give crude brown solid (1.7g), which was gradient elution was set as follows: 0–5 min (25% MeCN), 5–45min (25–55%
further purified by preparative HPLC with MeOH/0.1% HCO2H (MeOH MeCN). Retention times for the amino-acid standards were L-Ser-L-FDLA
concentration: 30% for 0–8min; 30–85% for 8–42min) at 12mlminꢀ1. The
22.0min and D-Ser-L-FDLA 22.9min, while the L-FDLA-hydrolysate of 1 gave a
fraction containing 1 was pooled, concentrated under reduced pressure, and the peak at 22.0 min.
remaining aqueous solution was lyophilized to yield catechoserine (1, tR
29.9 min, 13mg). Similarly, N,N¢-bis(2,3-dihydroxybenzoyl)-O-seryl serine (2,
tR 23.2min, 24mg) and N,N¢,N¢-tris(2,3-dihydroxybenzoyl)-O-seryl-O-seryl
serine (3, tR 29.9min, 20mg) were obtained from the HP-20 fractions eluted
with 60% and 80% MeOH, respectively.
Biological assays
Invasion and migration assays14 and cytotoxic, cell adhesion and matrix
metalloproteinase assays16 were carried out according to the procedures
previously described.
Catechoserine (1)
Colorless, amorphous solid; [a]2D3+13 (c 0.10, MeOH); UV (MeOH) lmax (log
e) 211 (4.62), 248 (4.17), 319 (3.78) nm; IR (ATR) nmax 3356, 2960, 1728,
;
1640 cmꢀ1 1H and 13C NMR data, see Table 1; HR-ESITOFMS (M+Na)+
320.1101 (calcd for C14H19NO6Na, 320.1105).
´
1
2
Berdy, J. Bioactive microbial metabolites. J. Antibiot. 58, 1–26 (2005).
Ayuso, A. et al. A novel actinomycete strain de-replication based on the diversity of
polyketide synthase and nonribosomal peptide synthetase biosynthetic pathways. Appl.
Microbiol. Biotechnol. 67, 795–806 (2005).
3
Gontang, E. A., Gaudencio, S. P., Fenical, W. & Jensen, P. R. Sequence-based analysis
of secondary-metabolite biosynthesis in marine actinobacteria. Appl. Environ. Micro-
biol. 76, 2487–2499 (2010).
Compound 2
Colorless, amorphous solid; [a]2D3+13 (c 0.10, MeOH); 1H NMR (500MHz,
DMSO-d6) d 3.78 (1H, dd, J¼11.0, 3.7Hz, H-8), 3.82 (1H, dd, J¼11.0, 5.5 Hz,
H-8), 4.41 (1H, dd, J¼11.0, 6.2 Hz, H-11), 4.57 (1H, m, H-9), 4.62 (1H, dd,
J¼11.0, 3.5 Hz, H-11), 4.78 (1H, m, H-12), 6.70 (1H, t, J¼7.5 Hz, H-5), 6.72
(1H, t, J¼7.5 Hz, H-19), 6.94 (1H, d, J¼7.0 Hz, H-4), 6.95 (1H, d, J¼7.0 Hz, H-
18), 7.33 (1H, d, J¼8.0 Hz, H-20), 7.37 (1H, d, J¼7.0 Hz, H-6), 8.94 (1H, d,
J¼7.5Hz, 7-NH), 8.97 (1H, br s, 12-NH); 13C NMR (125MHz, DMSO-d6)
d 51.7 (C-12), 55.2 (C-9), 60.7 (C-8), 63.7 (C-11), 115.5 (C-15), 115.9 (C-1),
118.1 (C-20), 118.21 (C-5)a 118.24 (C-19)a 118.6 (C-6), 118.8 (C-4), 118.9
(C-18), 146.0 (C-17), 146.1 (C-3), 148.2 (C-2), 148.6 (C-16), 168.5 (C-7), 168.7
(C-14), 169.9 (C-10), 170.3 (C-13); aassignment interchangeable. HR-ESI-
TOFMS (MꢀH)ꢀ 463.0988 (calcd for C20H19N2O11, 463.0944).
4
5
6
7
8
9
Goodfellow, M. & Fiedler, H. P. A guide to successful bioprospecting: informed by
actinobacterial systematics. Antonie Van Leeuwenhoek 98, 119–142 (2010).
Sato, S. et al. Indoxamycins A–F. Cytotoxic tricyclic polypropionates from a marine-
derived actinomycete. J. Org. Chem. 74, 5502–5509 (2009).
Iwata, F. et al. Lorneic acids, trialkyl-substituted aromatic acids from a marine-derived
actinomycete. J. Nat. Prod. 72, 2046–2048 (2009).
Ueda, J. et al. JBIR-58, a new salicylamide derivative, isolated from a marine sponge-
derived Streptomyces sp. SpD081030ME-02. J. Antibiot. 63, 267–269 (2010).
Igarashi, Y. et al. Pterocidin, a cytotoxic compound from the endophytic Streptomyces
hygroscopicus. J. Antibiot. 59, 193–195 (2006).
Igarashi, Y. et al. Alchivemycin A, a bioactive polycyclic polyketide with an unprece-
dented skeleton from Streptomyces sp. Org. Lett. 12, 3402–3405 (2010).
10 Igarashi, Y. et al. E. Abyssomicin I, a modified polycyclic polyketide from Streptomyces
sp. CHI39. J. Nat. Prod. 73, 1943–1946 (2010).
11 O’Brien, I. G. & Gibson, F. The structure of enterochelin and related 2,3-dihydroxy-
N-benzoylserine conjugates from Escherichia coli. Biochim. Biophys. Acta 215,
393–402 (1970).
Compound 3
¨
12 Kunze, B., Bedorf, N., Kohl, W., Hofle, G. & Reichenbach, H. Myxochelin A, a new iron-
chelating compound from Angiococcus disciformis (Myxobacterales). Production, iso-
lation, physico-chemical and biological properties. J. Antibiot. 42, 14–17 (1989).
13 Bhushan, R. & Bruckner, H. Marfey’s reagent for chiral amino acid analysis: a review.
Amino Acids 27, 231–247 (2004).
Colorless, amorphous solid; [a]2D3+11 (c 0.10, MeOH); 1H NMR (500MHz,
DMSO-d6) d 3.74 (1H, dd, J¼11.0, 3.7Hz, H-8), 3.80 (1H, dd, J¼11.0, 6.0 Hz,
H-8), 4.41 (1H, dd, J¼11.0, 7.0Hz, H-21), 4.44 (1H, dd, J¼11.0, 6.5 Hz, H-11),
4.59 (2H, m, H-9 and H-21), 4.61 (1H, dd, J¼11.0, 4.0 Hz, H-11), 4.74 (1H, br
s, H-12), 4.90 (1H, ddd, J¼7.0, 7.0, 4.5Hz, H-22), 6.67 (1H, t, J¼8.0 Hz, H-19),
6.68 (1H, t, J¼7.5 Hz, H-29), 6.69 (1H, t, J¼7.5 Hz, H-5), 6.93 (3H, m, H-4, H-
18, and H-28), 7.28 (1H, d, J¼8.0 Hz, H-30), 7.29 (1H, d, J¼8.0 Hz, H-20),
7.37 (1H, d, J¼8.0 Hz, H-6), 8.90 (1H, br s, 7-NH), 8.98 (1H, br s, 12-NH),
9.10 (1H, d, J¼7.0 Hz, 22-NH); 13C NMR (125MHz, DMSO-d6) d 52.0 (C-22),
52.3 (C-12), 55.6 (C-9), 61.3 (C-8), 63.6 (C-21), 64.7 (C-11), 115.8 (C-15)a
116.0 (C-25)a 116.6 (C-1), 118.5 (C-5)b 118.6 (C-19)b, 118.7 (C-20, C-29, C-
30)b,f, 119.1 (C-6), 119.3 (C-4)c, 119.4 (C-18)c, 119.5 (C-28)c, 146.54 (C-3)d,
146.56 (C-17)d, 146.58 (C-27)d, 148.7 (C-2), 149.2 (C-16)e, 149.4 (C-26)e,
168.8 (C-24), 169.1 (C-14), 169.3 (C-7), 169.5 (C-13), 170.3 (C-23), 170.8 (C-
10); aꢀeassignment interchangeable; foverlapped signals. HR-ESITOFMS
(MꢀH)ꢀ 686.1468 (calcd for C30H28N3O16, 686.1475).
14 Miyanaga, S. et al. Absolute configuration and antitumor activity of myxochelin A
produced by Nonomuraea pusilla TP-A0861. J. Antibiot. 59, 698–703 (2006).
15 Igarashi, Y. et al. Brartemicin, an inhibitor of tumor cell invasion from the actinomycete
Nonomuraea sp. J. Nat. Prod. 72, 980–982 (2009).
16 Miyanaga, S., Sakurai, H., Saiki, I., Onaka, H. & Igarashi, Y. Synthesis and evaluation of
myxochelin analogues as antimetastatic agents. Bioorg. Med. Chem. 17, 2724–2732
(2009).
17 Igarashi, Y. et al. Antitumor anthraquinones from an endophytic actinomycete Micro-
monosporalupini sp. nov. Bioorg. Med. Chem. Lett. 17, 3702–3705 (2007).
18 Neilands, J. B. Siderophores of bacteria and fungi. Microbiol. Sci. 1, 9–14 (1984).
19 Hantke, K., Nicholson, G., Rabsh, W. & Winkelmann, G. Salmochelins, siderophores
of Salmonella enterica and uropathogenic Escherichia coli strains, are recognized by
the outer membrane receptor IronN. Proc. Natl Acad. Sci. USA 100, 3677–3682
(2003).
20 Jiang, Y.- L. et al. Synthesis and evaluation of trehalose-based compounds as anti-
invasive agents. Bioorg. Med. Chem. Lett. 21, 1089–1091 (2011).
The Journal of Antibiotics