Abstract
Advances in genomics and metabolomics have made clear in recent years that microbial biosynthetic capacities on Earth far exceed previous expectations. This is attributable, in part, to the realization that most microbial natural product (NP) producers harbor biosynthetic machineries not readily amenable to classical laboratory fermentation conditions. Such "cryptic" or dormant biosynthetic gene clusters (BGCs) encode for a vast assortment of potentially new antibiotics and, as such, have become extremely attractive targets for activation under controlled laboratory conditions. We report here that coculturing of a Rhodococcus sp. and a Micromonospora sp. affords keyicin, a new and otherwise unattainable bis-nitroglycosylated anthracycline whose mechanism of action (MOA) appears to deviate from those of other anthracyclines. The structure of keyicin was elucidated using high resolution MS and NMR technologies, as well as detailed molecular modeling studies. Sequencing of the keyicin BGC (within the Micromonospora genome) enabled both structural and genomic comparisons to other anthracycline-producing systems informing efforts to characterize keyicin. The new NP was found to be selectively active against Gram-positive bacteria including both Rhodococcus sp. and Mycobacterium sp. E. coli-based chemical genomics studies revealed that keyicin's MOA, in contrast to many other anthracyclines, does not invoke nucleic acid damage.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3093-3102 |
| Number of pages | 10 |
| Journal | ACS Chemical Biology |
| Volume | 12 |
| Issue number | 12 |
| DOIs | |
| State | Published - Dec 15 2017 |
Bibliographical note
Funding Information:This work was supported by funding from the University of Wisconsin-Madison School of Pharmacy and from the University of Wisconsin Institute for Clinical and Translational Research funded through NIH/NCATS UL1TR000427. This work was also funded by the NIH through the administration of NIGMS Grants R01GM104192 (to T.S.B.) and R01GM104975 (to C.L.M.), NIDDK R01DK071801 (to L.L.), and HGRI Grants R01HG005084 (to C.L.M) and 1R01HG0050 (to C.L.M. and J.S.P.) as well as U19-Al109673 (to C.R.C.). NIH Biotechnology training grants T32GM008347 (to S.W.S.) and T32GM008505 (to M.C.) are also gratefully acknowledged. S.W.S. is also supported by an NSF Graduate Research Fellowship (00039202). We thank the Analytical Instrumentation Center at the University of Wisconsin-Madison for the facilities to acquire spectroscopic data. Orbitrap instruments were purchased through the support of an NIH shared instrument grant (NIH-NCRR S10RR029531 to L.L.) and the Office of the Vice Chancellor for Research and Graduate Education at the University of Wisconsin-Madison. This study made use of the National Magnetic Resonance Facility at Madison (NMRFAM) which is supported by NIH Grant P41GM103399. Additional equipment was purchased with funds from the University of Wisconsin, the NIH (RR02781, RR08438), the NSF (DMB-8415048, OIA-9977486, BIR-9214394) and the USDA. L.L. acknowledges a Vilas Distinguished Achievement Professorship and a Janis Apinis Professorship with funding provided by the Wisconsin Alumni Research Foundation and University of Wisconsin-Madison School of Pharmacy.
Funding Information:
This work was supported by funding from the University of WisconsinMadison School of Pharmacy and from the University of Wisconsin Institute for Clinical and Translational Research funded through NIH/NCATS UL1TR000427. This work was also funded by the NIH through the administration of NIGMS Grants R01GM104192 (to T.S.B.) and R01GM104975 (to C.L.M.), NIDDK R01DK071801 (to L.L.), and HGRI Grants R01HG005084 (to C.L.M) and 1R01HG0050 (to C.L.M. and J.S.P.), as well as U19-Al109673 (to C.R.C.). NIH Biotechnology training grants T32GM008347 (to S.W.S.) and T32GM008505 (to M.C.) are also gratefully acknowledged. S.W.S. is also supported by an NSF Graduate Research Fellowship (00039202). We thank the Analytical Instrumentation Center at the University of WisconsinMadison for the facilities to acquire spectroscopic data. Orbitrap instruments were purchased through the support of an NIH shared instrument grant (NIH-NCRR S10RR029531 to L.L.) and the Office of the Vice Chancellor for Research and Graduate Education at the University of WisconsinMadison. This study made use of the National Magnetic Resonance Facility at Madison (NMRFAM), which is supported by NIH Grant P41GM103399. Additional equipment was purchased with funds from the University of Wisconsin, the NIH (RR02781, RR08438), the NSF (DMB-8415048, OIA-9977486, BIR-9214394), and the USDA. L.L. acknowledges a Vilas Distinguished Achievement Professorship and a Janis Apinis Professorship with funding provided by the Wisconsin Alumni Research Foundation and University of WisconsinMadison School of Pharmacy. We also thank D. Demaria for assistance with sample collection and express our appreciation to H. Mori (Nara Institute of Science and Technology, Ikoma, Japan) for generous provision of the barcoded E. coli deletion collection.
Publisher Copyright:
© 2017 American Chemical Society.