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Antibacterial pyrrolidinyl and piperidinyl substituted 2,4-diacetylphloroglucinols from Pseudomonas protegens UP46

The Journal of Antibiotics volume 73, pages 739–747 (2020)Cite this article

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Abstract

In the search for new antibiotic compounds, fractionation of Pseudomonas protegens UP46 culture extracts afforded several known Pseudomonas compounds, including 2,4-diacetylphloroglucinol (DAPG), as well as two new antibacterial alkaloids, 6-(pyrrolidin-2-yl)DAPG (1) and 6-(piperidin-2-yl)DAPG (2). The structures of 1 and 2 were determined by nuclear magnetic resonance spectroscopy and mass spectrometry. Compounds 1 and 2 were found to have antibacterial activity against the Gram-positive bacteria Staphylococcus aureus and Bacillus cereus, with minimal inhibitory concentration (MIC) 2 and 4 μg ml−1, respectively, for 1, and 2 μg ml−1 for both pathogens for 2. The MICs for 1 and 2, against all tested Gram-negative bacteria, were >32 μg ml−1. The half maximal inhibitory concentrations against HepG2 cells for compounds 1 and 2 were 11 and 18 μg ml−1, respectively, which suggested 1 and 2 be too toxic for further evaluation as possible new antibacterial drugs. Stable isotope labelling experiments showed the pyrrolidinyl group of 1 to originate from ornithine and the piperidinyl group of 2 to originate from lysine. The P. protegens acetyl transferase (PpATase) is involved in the biosynthesis of monoacetylphloroglucinol and DAPG. No optical rotation was detected for 1 or 2, and a possible reason for this was investigated by studying if the PpATase may catalyse a stereo-non-specific introduction of the pyrrolidinyl/piperidinyl group in 1 and 2, but unless the PpATase can be subjected to major conformational changes, the enzyme cannot be involved in this reaction. The PpATase is, however, likely to catalyse the formation of 2,4,6-triacetylphloroglucinol from DAPG.

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Acknowledgements

Ultupharma AB is gratefully acknowledged for funding of JJL, JB, CN, BG, and AB.

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Authors and Affiliations

  1. Department of Molecular Sciences, Uppsala BioCenter, Swedish University of Agricultural Sciences, P.O. Box 7015, SE-750 07, Uppsala, Sweden

    Jolanta J. Levenfors, Christina Nord, Joakim Bjerketorp, Jerry Ståhlberg & Anders Broberg

  2. Ultupharma AB, Södra Rudbecksgatan 13, SE-752 36, Uppsala, Sweden

    Jolanta J. Levenfors, Joakim Bjerketorp & Bo Öberg

  3. Department of Medical Sciences, Uppsala University, SE-751 85, Uppsala, Sweden

    Rolf Larsson

  4. Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, P.O. Box 7036, SE-750 07, Uppsala, Sweden

    Bengt Guss

  5. Department of Medicinal Chemistry, Uppsala University, P.O. Box 574, SE-751 23, Uppsala, Sweden

    Bo Öberg

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Levenfors, J.J., Nord, C., Bjerketorp, J. et al. Antibacterial pyrrolidinyl and piperidinyl substituted 2,4-diacetylphloroglucinols from Pseudomonas protegens UP46. J Antibiot 73, 739–747 (2020). https://doi.org/10.1038/s41429-020-0318-1

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