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Nature Synthesis volume 3, pages 99–110 (2024 )Cite this article Pentane Cyclopentane
Naturally occurring β-lactone compounds are a class of strained four-membered heterocycles and can act as highly reactive electrophiles. Despite intensive studies for several decades, fewer than 30 β-lactones, many of which have high clinical potential, have been characterized from microorganisms. Here we report the discovery of a β-lactone compound, globilactone A through heterologous expression of a polyketide synthase/non-ribosomal peptide synthetase-like biosynthetic gene cluster (glo) in Streptomyces albus J1074. Biosynthetic pathway studies revealed that the polyketide synthase part can synthesize a polyunsaturated polyketide chain. While the downstream non-ribosomal peptide synthetase-like module, comprising condensation, FkbH, peptidyl carrier protein and thioester reductase domains (C–FkbH–PCP–R), incorporates a three-carbon pyruvate unit, and mediates formation of two carbon–carbon bonds between the polyketide and pyruvate to give a cyclopentane intermediate tethered on acyl carrier protein. A downstream esterase, GloD, plays a direct role in the β-lactone ring formation and releases the cyclopentane–β-lactone from the assembly line.
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Dale F. Kreitler, Erin M. Gemmell, … Andrew M. Gulick
Joleen Masschelein, Pauline K. Sydor, ... Gregory L. Challis
Asfandyar Sikandar, Laura Franz, … Jesko Koehnke
Data supporting the findings of this work are available within the paper and its Supplementary Information files. The DNA sequence of gene cluster glo has been deposited in GenBank with the accession number OP620071 (https://www.ncbi.nlm.nih.gov/nuccore/OP620071.1/).The source data underlying Figs. 4a–d and 5c,d; Extended Data Figs. 2, 6a,b, 8a–c, 9a–c and 10a,b are provided as a Source Data file. Source data are provided with this paper.
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This research was financially supported by MOST (2022YFC2303100, 2022YFC2804100 and 2018YFA0902000), NSFC (81925033, 22193071, 22107048, 22077062, 81991522 and 81991524). We thank H. Zheng (China Pharmaceutical University) for his generous gift of expression plasmid NDM-1.
State Key Laboratory of Pharmaceutical Biotechnology, Department of Neurology, Nanjing Drum Tower Hospital, School of Life Sciences, Chemistry and Biomedicine Innovation Center (ChemBIC), Nanjing University, Nanjing, China
Zi Fei Xu, Ye Lei Zhou, Sheng Tao Bo, Jing Shi, Lang Xiang, Meng Yu Xi, Bo Zhang, Ren Xiang Tan & Hui Ming Ge
State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, China
Shou Qi Zhang & Zheng Ren Xu
Department of Chemistry, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation and Technology, University of Florida, Jupiter, FL, USA
Dong Yang & Ben Shen
Natural Products Discovery Center, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation and Technology, University of Florida, Jupiter, FL, USA
Dong Yang & Ben Shen
Department of Molecular Medicine, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation and Technology, University of Florida, Jupiter, FL, USA
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Z.F.X., Y.L.Z. and S.T.B. carried out experiments. L.X., M.Y.X., J.S., B.Z. and R.X.T. assisted in NMR and MS data measurement and analysis. S.Q.Z., Z.R.X., D.Y. and B.S. contributed materials. Z.F.X. and H.M.G. wrote the paper. H.M.G. supervised the work. All authors discussed the results and analysed the data.
Correspondence to Ren Xiang Tan or Hui Ming Ge.
The authors declare no competing interests.
Nature Synthesis thanks the anonymous reviewers for their contribution to the peer review of this work. Primary Handling Editor: Thomas West, in collaboration with the Nature Synthesis team.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
cis-3-octyl-4-nonyloxetan-2-one (a), obafluorin (b), vibralactone (c), salinosporamide A (d).
An IC50 value against class A β-lactamase of 1 was 15.05 μM.
a Reaction scheme for the GloE activity assay. b HPLC analysis of the enzymatic assays of GloE with 4. c HPLC analysis of the enzymatic assays of GloE with 1.
Heavy bars denote contiguous 13C labels of connected atoms, dots denote single 13C labels.
Reaction scheme depicting chemoenzymatic synthesis and subsequent acylation assay to generate 8-S-ACP (GloB), 8-S-ACP-C-FkbH (GloB) (a) and 13-S-GloC-GloD (b).
a LC-MS analysis of the one-pot enzymatic total biosynthesis of 10. i) boiled holo-GloC-GloD, 8-S-ACP-C-FkbH (GloB), 1,3-BPG, NADPH and N-acetyl-L-cysteine; ii) holo-GloC-GloD, 8-S-ACP-C-FkbH (GloB), 1,3-BPG and N-acetyl-L-cysteine; iii) holo-GloC-GloD, 8-S-ACP-C-FkbH (GloB), 1,3-BPG, NADPH and N-acetyl-L-cysteine. b high-resolution MS (HRMS) data of 10.
Reaction scheme depicting chemoenzymatic synthesis and subsequent acylation assay to generate 11-S-ACP-C-FkbH (GloB).
a holo-ACP (GloB). b ACP-tethered linear substrate (8-S-ACP) from pantetheine-activated (8c) via in vitro CoA biosynthesis and Sfp-mediated loading. c holo-GloC-GloD, holo-ACP-C-FkbH (GloB), 1,3-BPG, NADPH and 8-S-ACP. Additional peaks marked by asterisks represent adventitious gluconylation (+178 Da) of N-terminal His6-tag during expression of the recombinant protein in E. coli.
a holo-ACP (GloB). b ACP-tethered linear substrate (8-S-ACP) from pantetheine-activated (8c) via in vitro CoA biosynthesis and Sfp-mediated loading. c holo-GloC-GloD, holo-ACP-C-FkbH (GloB), 1,3-BPG, NADPH and 8-S-ACP.
a LC-MS analysis of the one-pot enzymatic total biosynthesis of 3. b Mass spectrometric analysis of production of acylated ACP (12). Additional peaks marked by asterisks represent adventitious gluconylation (+178 Da) of N-terminal His6-tag during expression of the recombinant protein in E. coli.
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Xu, Z.F., Zhou, Y.L., Bo, S.T. et al. Discovery and biosynthetic pathway analysis of cyclopentane–β-lactone globilactone A. Nat. Synth 3, 99–110 (2024). https://doi.org/10.1038/s44160-023-00414-3
DOI: https://doi.org/10.1038/s44160-023-00414-3
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