Letter

Cite This: Org. Lett. 2018, 20, 1563−1567 pubs.acs.org/OrgLett

Georatusin, a Specific Antiparasitic Polyketide−Peptide Hybrid from

the Fungus Geomyces auratus
Yi-Ming Shi,† Christian Richter,‡ Victoria L. Challinor,† Peter Grün,† Antonio Girela del Rio,§
Marcel Kaiser,⊥ Anja Schüffler,# Meike Piepenbring,§ Harald Schwalbe,‡ and Helge B. Bode*,†,∥
†
Merck-Stiftungsprofessur für Molekulare Biotechnologie, Fachbereich Biowissenschaften, ‡Institut für Organische und Chemische
Biologie, Zentrum für Biomolekulare Magnetische Resonanz, §Department of Mycology, Fachbereich Biowissenschaften, Biologicum,
and ∥Buchmann Institute for Molecular Life Sciences (BMLS), Goethe Universität Frankfurt, 60438 Frankfurt am Main, Germany
⊥
Swiss Tropical and Public Health Institute Parasite Chemotherapy and University of Basel, 4051 Basel, Switzerland
#
See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles.

Institut für Biotechnologie und Wirkstoff-Forschung gGmbH (IBWF), 67663 Kaiserslautern, Germany
*
S Supporting Information
Downloaded via UNIV FRANKFURT on October 24, 2018 at 06:42:40 (UTC).

ABSTRACT: Georatusin (1), featuring a highly reduced, methylated

polyketide moiety fused to a tryptophan by an amide and ester bond forming
a 13-membered ring, was produced by the soil fungus Geomyces auratus. An
HMQC−COSY spectrum was measured to build up the connectivities despite
the overlapping proton signals. DQF-COSY, HETLOC, J-HMBC, and ROESY
were implemented to determine the relative configuration of the flexible
moiety. Georatusin (1) shows specific antiparasitic activities against Leishmania
donovani and Plasmodium falciparum without obvious cytotoxicity. The
biosynthesis of 1 was also proposed.

F ungi make up a taxon of eukaryotic organisms with

probably more than 1.5 million estimated species,1
diverging from the other two eukaryotic kingdoms (animals
and plants). Living in multispecies communities, fungi produce
small molecules as antibiotics and signaling compounds for
intercellular communication,2 interacting with the host and
other microorganisms, and defeating other microbial com-
petitors to thrive in complex ecosystems.3 The vast repertoire
of fungal natural products with their unusual modes of action4
has inspired generations of researchers to explore new chemical
entities and search for small-molecule probes and therapeutic
agents. This includes the penicillin antibiotics, the immuno-
suppressant cyclosporin, and the cholesterol-reducing drug
lovastatin.4,5 However, despite the high value of the identified
natural products and the wide occurrence of fungi in all
different environments, <10% of fungal species are known
taxonomically and chemically.1
As part of exploring and exploiting the potential of fungi in
the Integrative Fungal Research (IPF) initiative, we isolated Figure 1. (A) Structure of georatusin (1). (B) Connectivities of 1
based on 1H−1H COSY (bold red line), HMQC−COSY (bold green
Geomyces auratus, a mesophilic fungus (Figures S1 and S2)
line), and HMBC (from H to C, blue arrow) data.
belonging to the family Myxotrichaceae, from a soil sample
collected in Germany. After cultivation on potato dextrose agar
at room temperature for 2 weeks, the agar plates were extracted reduced, methylated polyketide moiety fused to tryptophan
with ethyl acetate. The UPLC−UV/MS chromatogram (Figure by an amide and ester bond forming a 13-membered ring.
S3) showed four major peaks in which the least polar one (tR = Herein, we report its structural elucidation by two-dimensional
11.3 min) showed a signal of m/z 551.3802 [M − H2O + H]+, (2D) NMR, including J-based configuration analysis (JBCA)
being much larger than the other three compounds. To identify for the establishment of the relative configuration, and propose
the structure of this hydrophobic compound, we subjected the its biosynthesis pathway.
organic layer to column chromatography on silica gel and
semipreparative HPLC to afford georatusin [1 (Figure 1A)], a Received: January 27, 2018
polyketide−peptide hybrid metabolite featuring a highly Published: February 23, 2018

© 2018 American Chemical Society 1563 DOI: 10.1021/acs.orglett.8b00293

Org. Lett. 2018, 20, 1563−1567
Organic Letters
Georatusin (1) was obtained as a colorless gum, and positive-
ion HRESIMS provided a formula of C34H52N2O5 from an ion
peak at m/z 591.3757 [M + Na] + (calculated for
C34H52N2O5Na, m/z 591.3768, Δppm 2.0), requiring 10
degrees of unsaturation. Inspection of the 1H NMR spectrum
of 1 recorded in pyridine-d5 (Table S1 and Figure S4) showed
two secondary methyls at δH 0.70 (J = 6.5 Hz) and 1.23 (J = 7.1
Hz), as well as six overlapping methyls. Nine one-proton
multiplets (δH 1.61, 1.77, 2.07, 2.48, 2.75, 3.61, 3.92, 4.11, and
4.83) were assigned to either methines or methylenes. In the
low-field region, two singlets at δH 7.09 and 11.93, four
doublets at δH 7.48 (J = 1.9 Hz), 7.60 (J = 8.0 Hz), 7.88 (J = 8.0
Hz), and 9.57 (J = 7.7 Hz), and one triplet at δH 7.26 (J = 7.5
Hz) resonated from either olefinic or exchangeable protons.
Examination of 2D NMR spectra, including HSQC (Figures
S6 and S13), HMBC (Figures S7 and S14), 1H−1H COSY
(Figure S8), and HMQC−COSY (Figure S9), allowed the
planar structure assignment (Figure 1B). The HSQC spectrum
recorded in pyridine-d6 (Figure S6) enabled the assignment of
all protons to the directly bonded carbons, except for three
signals at δH 7.09, 9.57, and 11.9, indicating that these three
were exchangeable protons. The relative low-field chemical
shifts of 9.57 and 11.9 ppm along with two 1H−1H COSY spin
systems coupling with δH 4.83 and 7.48, respectively, enabled
the assignment of two NH protons,6 rationalizing that the
exchangeable proton at δH 7.09 was from a hydroxyl group.
An indole ring can be readily deduced by the characteristic
13
C chemical shifts (Table S1)6 and was supported by analysis
of the 1H−1H COSY spectrum (Figure S8). Two proton signals
at δH 7.88 (H-4) and 11.90 (NH-1) showed H-4/H-5/H-6/H-7
and NH-1/H-2 spin systems, respectively, and HMBC (Figure
S7) showed correlations from NH-1 to C-3a (δC 128.2) and C-
3 (δC 111.7), and from H-2 and H-4 to C-3a, C-3, and C-7a (δC
137.4). Three proton signals appearing as an ABX spin system
at δH 3.92 (dd, J = 14.4, 7.4 Hz), 4.11 (dd, J = 14.4, 8.4 Hz),
and 4.83 (br dd, J = 9.1, 8.4 Hz) could be readily assigned to β-
proton (H-8a and H8-b) and α-proton (H-9) in a tryptophan
moiety (part A, Figure 1B).
Two methyl doublets at δH 0.85 and 0.70 showed HMBC
(Figure S7) correlations with an oxygenated quaternary carbon
at δC 99.1 (C-14) and an oxymethine at δC 77.5 (C-18),
respectively, as well as shared a correlation with a methylene at
δC 35.7 (C-16). A further 1H−1H COSY spin system of
−CH3CHCH2CH(CH3)CH−, in a combination of a key
HMBC correlation from H-18 to C-14, indicated the presence
of a six-membered oxygen-containing ring. The hydroxyl group
(δH 7.09) was located at C-14, which was judged by the HMBC
correlations from OH-14 to C-14, C-15, and a methine at δH
2.75 (C-13), revealing C-14 to be a hemiketal. A methyl
doublet at δH 1.23 (C-28) showed HMBC correlations with
both a carbonyl group at δC 177.3 (C-12) and C-14 and had a
1
H−1H COSY correlation with a quartet methine at δH 2.75
(C-13), indicating C-12 attaching to C-14 through C-13. Thus,
another substructure (part B, Figure 1B) was established.
The number of the splitting methyls ranging from δH 0.79 to
0.84 was equal to that of the remaining methines, suggesting
that each of these methyls might be linked to a methine.
However, the possibility that one of these methyls was attached
to the methine at δC 80.8 could be excluded by the absence of a
1
H−1H COSY between the oxymethine and any methyl.
Therefore, it was assumed that one of the five methyls was
connected to a methylene as a terminal methyl. 1H−1H COSY
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Letter
failed to establish a defined substructure due to the overlapping
proton signals in the high-field region. Then, an HMQC−
COSY spectrum (Figure S9) was measured to detect the
relayed carbon connectivity, coupling with the HMBC data,
which led to a polyketide spin system (part C, Figure 1B) with
an oxygenated substitution at C-22.
The assembly of parts A and B was facilitated by the
simultaneous HMBC correlations (Figure S7) from NH-11 and
H-9 to C-12, suggesting these two parts are linked through an
amide bond. The 1H−1H COSY (Figure S8) correlation of H-
18/H-19 and HMBC correlation from Me-31 to C-18
furnished a σ-bond connection between parts B and C. From
an HMBC correlation of an oxymethine (H-22) with a
carboxylic group (C-10) a linkage between parts A and C by
an ester bridge was determined. Thus, the planar structure of a
polyketide−peptide hybrid was established (Figure 1B).
The C-9 stereogenic center in tryptophan of 1 could be
readily determined by an empirical comparison of its electronic
circular dichroism (ECD) spectrum with those of D- and L-
tryptophan. The ECD curve of 1 showed a diagnostic negative
Cotton effect at 238 nm (Figure S15), consistent with that of D-
tryptophan.
With respect to the relative configuration of the tetrahy-
dropyran moiety and its peripheral stereogenic centers, two
large couplings 3JH‑15,H‑16b (10.2 Hz) and 3JH‑17,H‑18 (8.3 Hz) and
a small coupling 3JH‑18,H‑19 (2.2 Hz) indicated that H-15/H-16b,
H-17/H-18, and H-18/H-19 are anti, anti, and gauche
orientations, respectively, as determined in the DQF-COSY
spectrum (Figure S16). The overlapping proton signals
obscured the measurement of coupling constants between H-
16a and H-16b with H-17; nevertheless, H-18 showed ROESY
(Figure S17) cross peaks with OH-14 and H-16a, supporting
the finding that they are axially oriented, which was further
defined as β-oriented. H-13, C-29, and H-17 are α-oriented,
which were determined by the ROESY correlations of H-13/
Me-29/H-17 and H-15/Me-28. Thus, the stereogenic centers
are established to be 13R*, 14R*, 15S*, 17S*, and 18R*
(Figure 2).
Figure 2. Key ROESY correlations for the tetrahydropyran moiety and
its peripheral stereocenters.
The relative configuration of the flexible polyketide moiety
with five stereogenic centers could be established by using
JBCA7 as well as a ROESY spectrum (Figure S20). The C-18/
C-19 relative was established by examining the measured 3JH,H
in DQF-COSY (Figure S16) and 2,3JC,H in HETLOC (Figure
S18) and J-HMBC (Figure S19) spectra. The small 3JH,H of H-
18/H-19 (2.2 Hz) accounted for a gauche relationship. 3JC,H
values of H-18/C-20 (2.6 Hz) and C-31/H-18 (5.8 Hz) placed
H-18 gauche and anti for C-20 and C-31, together with a small
2
JC,H value of C-18/H-19 (1.3 Hz), which led to the assignment
of 19S*. The observations of 3JH,H values of 3.7 and 8.9 Hz for
H-20a/H-21 and H-20b/H-21, respectively, supported gauche
DOI: 10.1021/acs.orglett.8b00293
Org. Lett. 2018, 20, 1563−1567
Organic Letters
and anti orientations, respectively. Two small 3JC,H values for H-
20a/C-22 (2.5 Hz) and H-20b/C-22 (2.7 Hz) placing C-22
gauche concerning H-20a and H-20b along with a large 3JC,H for
C-32/H-20a (8.0 Hz) led to the determination of 21S*. For the
C-21/C-22 fragment, a 3JH,H value of 10.4 Hz for H-21/H-22
supported an anti relationship. Two small 3JC,H couplings for C-
20/H-22 (1.9 Hz) and C-32/H-22 (2.7 Hz) indicated their
gauche orientations. The ester oxygen must be positioned
gauche to H-21 based on a large coupling for C-22 and H-21
(−6.5 Hz). A ROESY correlation that was observed for H-23/
H-32 facilitated the identification of the only possible
diastereomer, enabling the identification of 22R*. H-22 showed
two small couplings with H-23 (1.5 Hz) and C-24 (3.5 Hz), as
well as a large coupling with C-33 (5.6 Hz), supporting the
assignment of 23S*. With regard to the stereochemical
relationship between C-23 and C-24, two medium values of
8.8 and 5.3 Hz were observed for H-23/H-24a and H-23/H-
24b, respectively, suggesting the existence of two alternating
rotamers. The small 3JC,H values of C-22/H-24a (1.2 Hz) and
C-33/H-24b (2.4 Hz) indicated their gauche orientations, and
therefore, a single pair of rotamers met these requirements. C-
25 was determined to be R*, which was supported by the
gauche and anti relationships for H-24a/H-25 (3.9 Hz) and H-
24b/H-25 (9.5 Hz), respectively, together with the anti
placement (6.2 Hz) of C-34 with respect to H-24a and the
gauche relationship of C-34/H-24b (2.9 Hz). As a result, the
stereogenic centers of the flexible moiety are established to be
19S*, 21S*, 22R*, 23S*, and 25R* (Figure S20).
The challenges of the structural interpretation of 1 stem from
the polyketide motif. (1) The overlapping methyl signals in the
high-field region of the 1H NMR spectrum are ineffective for
1
H−1H COSY in combination with HMBC, preventing the
determination of the connectivities of the motif. (2) The
ROESY-based technique for the relative configurational
assignment of the five stereogenic centers (C-19, C-21, C-22,
C-23, and C-25) is hampered by the flexible carbon chain (from
C-19 to C-27) with multiple conformers. Therefore, to address
the overlapping signal issue in 1H−1H COSY, we performed an
HMQC−COSY experiment8 on a 700 MHz NMR instrument
to provide highly dispersed 2JH,C correlations that retain the
significant information regarding pattern connectivity via
1
H−1H COSY.
The determination of the relative configuration in the flexible
chain was achieved by a combination of DQF-COSY,
HETLOC, and J-HMBC, termed JBCA,7 which has been
widely applied to acyclic and macrolide structures.9
It was reported that one Geomyces species, Geomyces
destructans, is the causal agent of the white-nose syndrome of
bats that results in high bat mortality.10 Although Geomyces
species were subsequently reported in soil from bat hibernacula,
their pathogenicities and ecological relationships remained
cryptic.11 So far, little has been discovered about the metabolic
profiles of Geomyces species in general, and only eight asterric
acid derivatives12 and pannomycin13 have been described.
Georatusin (1) shows antiparasitic activities against Leishma-
nia donovani (IC50 = 9.1 μM) and Plasmodium falciparum [IC50
= 1.6 μM (Table 1)] without antibacterial and antifungal
activity. Interestingly, metacridamides A and B (Scheme 1A),
structurally similar to 1,14 show cytotoxicity toward several
cancer cell lines, while 1 is not cytotoxic, thus presumably
pointing to the α,β-unsaturated amide in metacridamides as a
Michael receptor contributing to their cytotoxicity.
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Letter
Table 1. Bioactivities of 1 against Different Protozoa and
Cytotoxicities (IC50 in micromolar)
species 1 controla
Trypanosoma brucei rhodesiense 56.3 0.005
Trypanosoma cruzi >100 2.7
L. donovani 9.1 1.2
P. falciparum 1.6 0.008
mammalian L6 cells >100 0.008
a
The controls differ for each tested organism. Melarsoprol,
benznidazole, miltefosine, chloroquine, and podophyllotoxin were
used as controls for T. brucei rhodesiense, T. cruzi, L. donovani, P.
falciparum, and mammalian L6 cells, respectively.
Because of the pathogenicity of G. destructans and the
widespread occurrence of the Geomyces genus, not only in bat
hibernacula but also commonly in soil and cool environ-
ments,10 it is tempting to speculate that the specific antiparasitic
function of georatusin (1) might be essential to help the fungi
establish their ecological niche. The discovery of the unique
structure and specific bioactivity of 1 provides new insight into
the metabolic capabilities and ecological significance of
Geomyces and clearly warrants the exploration of this genus in
the future.
With respect to the biosynthesis of highly reduced
polyketide−peptide hybrids from fungi, compounds such as
aspyridone A,15 cytochalasan,16 and thermolide A (Scheme
1A)17 have been described in the past decade. Because no
Geomyces genomes are available in the NCBI database, the
biosynthetic pathway of georatusin [1 (Scheme 1B)] was
proposed on the basis of the established paradigms17−20 and
the genome sequences of Metarhizium species, among which
Metarhizium acridium is a known producer of metacrida-
mides.14 Antismash analysis21 of genome sequences from 11
Metarhizium species led to the identification of at least 16
biosynthesis gene clusters encoding two different polyketide−
peptide synthases (Figure S21).22 The clusters encode either a
single polyketide−peptide megasynthetase hybrid with a
terminal R domain and a trans-acting ER (Figure S21, type I)
or two separate enzymes encoding a PKS and an NRPS with a
C-terminal domain (Figure S21, type II). Consequently, there
are two conceivable pathways for forming the macrolide of
georatusin (1). (1) The R domain reductively releases an
amino aldehyde that is attacked by OH-22 followed by
oxidation (pathway a, Scheme 1B), as presented in the
lactonization of thermolide.17 (2) The C-terminal domain off-
loads the thioester-tethered amide via cyclization to form the
lactone (pathway b, Scheme 1B).19 Another interesting feature
of the proposed biosynthesis of 1 is the recruitment of a D-
amino acid rather than incorporation of an L-amino acid, in
contrast to the case for metacridamides. To the best of our
knowledge, thermolide analogues from thermophilic fungi17,20
are the only other examples for the occurrence of a D-amino
acid in such natural products. However, the mechanism for D-
amino acid selection or epimerization has not yet been
elucidated. It might be possible that the A domain is
responsible for the exclusive incorporation of a D-tryptophan
provided by an epimerase encoded elsewhere in the genome.
DOI: 10.1021/acs.orglett.8b00293
Org. Lett. 2018, 20, 1563−1567
Organic Letters Letter

Scheme 1. (A) Known Highly Reduced Polyketide−Peptide Hybrids from Fungi Exemplified by Thermolide A, Metacridamide
A, and Aspyridone A and (B) Proposed Biosynthetic Pathway for Georatusin (1)a

a
ER may be defective and complemented by a trans-acting ER as shown in Figure S21 (type I).

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*
ASSOCIATED CONTENT
S Supporting Information
(4) (a) Keller, N. P.; Turner, G.; Bennett, J. W. Nat. Rev. Microbiol.
2005, 3, 937−947. (b) Schueffler, A.; Anke, T. Nat. Prod. Rep. 2014,
31, 1425−1448.
The Supporting Information is available free of charge on the (5) Brakhage, A. A.; Schroeckh, V. Fungal Genet. Biol. 2011, 48, 15−
ACS Publications website at DOI: 10.1021/acs.or- 22.
glett.8b00293. (6) Grundmann, F.; Kaiser, M.; Schiell, M.; Batzer, A.; Kurz, M.;
Thanwisai, A.; Chantratita, N.; Bode, H. B. J. Nat. Prod. 2014, 77,
Detailed experimental procedures, NMR and ECD 779−783.
spectra, and PKS-NRPS biosynthetic gene clusters from (7) Matsumori, N.; Kaneno, D.; Murata, M.; Nakamura, H.;
Metarhizium (PDF) Tachibana, K. J. Org. Chem. 1999, 64, 866−876.

■
(8) Hu, K.; Westler, W. M.; Markley, J. L. J. Biomol. NMR 2011, 49,
AUTHOR INFORMATION 291−296.
(9) (a) Bifulco, G.; Dambruoso, P.; Gomez-Paloma, L.; Riccio, R.
Corresponding Author Chem. Rev. 2007, 107, 3744−3779. (b) Kawahara, T.; Izumikawa, M.;
*E-mail: h.bode@bio.uni-frankfurt.de. Takagi, M.; Shin-Ya, K. Org. Lett. 2012, 14, 4434−4437. (c) Sikorska,
J.; Hau, A. M.; Anklin, C.; Parker-Nance, S.; Davies-Coleman, M. T.;
ORCID Ishmael, J. E.; McPhail, K. L. J. Org. Chem. 2012, 77, 6066−6075.
Yi-Ming Shi: 0000-0001-6933-4971 (d) Rodriguez, J.; Nieto, R. M.; Blanco, M.; Valeriote, F. A.; Jimenez,
Harald Schwalbe: 0000-0001-5693-7909 C.; Crews, P. Org. Lett. 2014, 16, 464−467.
(10) Lorch, J. M.; Meteyer, C. U.; Behr, M. J.; Boyles, J. G.; Cryan, P.
Helge B. Bode: 0000-0001-6048-5909
M.; Hicks, A. C.; Ballmann, A. E.; Coleman, J. T.; Redell, D. N.;
Notes Reeder, D. M.; Blehert, D. S. Nature 2011, 480, 376−378.
The authors declare no competing financial interest. (11) Lorch, J. M.; Lindner, D. L.; Gargas, A.; Muller, L. K.; Minnis, A.

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ACKNOWLEDGMENTS
Y.-M.S. is supported by a Postdoctoral Research Fellowship
from the Alexander von Humboldt Foundation. This work was
funded in part by the state of Hesse as part of the LOEWE
research cluster Integrated Fungal Research (IPF). The authors
thank the following colleagues from the Goethe Universität
Frankfurt: Miguel Rosas for isolating the fungal strain, Jascha
Weisenborn for providing a preliminary identification, Florian
Hennicke for managing the strain collection, and Dr. Krishna
Saxena for ECD measurement.
M.; Blehert, D. S. Mycologia 2013, 105, 237−252.
(12) Li, Y.; Sun, B.; Liu, S.; Jiang, L.; Liu, X.; Zhang, H.; Che, Y. J.
Nat. Prod. 2008, 71, 1643−1646.
(13) Parish, C. A.; de la Cruz, M.; Smith, S. K.; Zink, D.; Baxter, J.;
Tucker-Samaras, S.; Collado, J.; Platas, G.; Bills, G.; Diez, M. T.;
Vicente, F.; Peláez, F.; Wilson, K. J. Nat. Prod. 2009, 72, 59−62.
(14) Krasnoff, S. B.; Englich, U.; Miller, P. G.; Shuler, M. L.; Glahn,
R. P.; Donzelli, B. G.; Gibson, D. M. J. Nat. Prod. 2012, 75, 175−180.
(15) Bergmann, S.; Schumann, J.; Scherlach, K.; Lange, C.; Brakhage,
A. A.; Hertweck, C. Nat. Chem. Biol. 2007, 3, 213−217.
(16) Schümann, J.; Hertweck, C. J. Am. Chem. Soc. 2007, 129, 9564−
9565.

■
REFERENCES
(1) Hawksworth, D. L. Mycol. Res. 2001, 105, 1422−1432.
(2) (a) Hornby, J. M.; Jensen, E. C.; Lisec, A. D.; Tasto, J. J.; Jahnke,
B.; Shoemaker, R.; Dussault, P.; Nickerson, K. W. Appl. Environ.
Microbiol. 2001, 67, 2982−2992. (b) Hogan, D. A. Eukaryotic Cell
2006, 5, 613−619.
(3) Kusari, S.; Hertweck, C.; Spiteller, M. Chem. Biol. 2012, 19, 792−
798.
(17) Niu, X.; Chen, L.; Yue, Q.; Wang, B.; Zhang, J.; Zhu, C.; Zhang,
K.; Bills, G. F.; An, Z. Org. Lett. 2014, 16, 3744−3747.
(18) Boettger, D.; Hertweck, C. ChemBioChem 2013, 14, 28−42.
(19) (a) Gao, X.; Haynes, S. W.; Ames, B. D.; Wang, P.; Vien, L. P.;
Walsh, C. T.; Tang, Y. Nat. Chem. Biol. 2012, 8, 823−830. (b) Zhang,
J.; Liu, N.; Cacho, R. A.; Gong, Z.; Liu, Z.; Qin, W.; Tang, C.; Tang,
Y.; Zhou, J. Nat. Chem. Biol. 2016, 12, 1001−1003.
(20) Guo, J. P.; Zhu, C. Y.; Zhang, C. P.; Chu, Y. S.; Wang, Y. L.;
Zhang, J. X.; Wu, D. K.; Zhang, K. Q.; Niu, X. M. J. Am. Chem. Soc.
2012, 134, 20306−20309.

1566 DOI: 10.1021/acs.orglett.8b00293

Org. Lett. 2018, 20, 1563−1567
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(21) Blin, K.; Wolf, T.; Chevrette, M. G.; Lu, X.; Schwalen, C. J.;
Kautsar, S. A.; Suarez Duran, H. G.; de los Santos, E. L. C.; Kim, H. U.;
Nave, M.; Dickschat, J. S.; Mitchell, D. A.; Shelest, E.; Breitling, R.;
Takano, E.; Lee, S. Y.; Weber, T.; Medema, M. H. Nucleic Acids Res.
2017, 45, W36−W41.
(22) (a) Sbaraini, N.; Guedes, R. L.; Andreis, F. C.; Junges, A.; de
Morais, G. L.; Vainstein, M. H.; de Vasconcelos, A. T.; Schrank, A.
BMC Genomics 2016, 17, 736. (b) Gao, Q.; Jin, K.; Ying, S.-H.; Zhang,
Y.; Xiao, G.; Shang, Y.; Duan, Z.; Hu, X.; Xie, X.-Q.; Zhou, G.; Peng,
G.; Luo, Z.; Huang, W.; Wang, B.; Fang, W.; Wang, S.; Zhong, Y.; Ma,
L.-J.; St. Leger, R. J.; Zhao, G.-P.; Pei, Y.; Feng, M.-G.; Xia, Y.; Wang,
C. PLoS Genet. 2011, 7, e1001264.

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