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Putisolvin I-II

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General info

Original publicationKuiper, 2003
Original sourcePseudomonas putida PCL1445
Other known sources (non-putative)Pseudomonas putida 267 (Kruijt, 2009)
Pseudomonas putida E41 (Bernat, 2019)
Pseudomonas capeferrum HN2-3 (Sheng, 2024)
Pseudomonas sp. COR55 (Muangkaew, 2024)
Pseudomonas vlassakiae WCU_60 (Muangkaew, 2024)
Pseudomonas vlassakiae WCU_64 (Oni, 2020; Muangkaew, 2024)
Pseudomonas capeferrum WCS358T (Muangkaew, 2024)
Pseudomonas sp. NNC7(Oni, 2020; Muangkaew, 2024)
Pseudomonas fulva LMG 11722T (Muangkaew, 2024)
Pseudomonas sp. COR19 (Oni, 2020; Muangkaew, 2024)
Stereochemistry determined byChemical synthesis and NMR fingerprinting (Muangkaew, 2024)

Chemical properties

CASn.a.
Molecular formulaPutisolvin I: C65H113N13O19
Putisolvin II: C66H115N13O19
Molecular weightPutisolvin I: 1380.7 g/mol
Putisolvin II: 1394.7 g/mol
Mono-isotopic massPutisolvin I: 1379.8276 Da
Putisolvin II: 1393.8432 Da
Solubilityn.a.
CMCn.a.
Minimal surface tensionn.a.
3D conformationn.a.
NMR data available in literaturen.a.

Introduction

Putisolvins are cyclic lipodepsipeptides produced by many different soil-dwelling Pseudomonas bacteria. were first described in the context of a study into the biosurfactant properties of bacteria grown on soils polluted with polycyclic aromatic hydrocarbons oil-contaminated soils. More specifically, it was observed that the original putisolvin-producing strain, Pseudomonas putida PCL1445, produces biosurfactant activity which was traced back to the production of two structurally related lipopeptides, putisolvin I and II.

Chemical structure

The structure of putisolvin I was initially determined by tandem mass spectrometry and 1H NMR spectroscopy. The sequence determined by means of MS/MS was complemented by NMR sequential data obtained from ROESY spectra, including those of the isobaric Leu/Ile residues. (Kuiper, 2003) A later study supported the structures of putisolvin I and II by the analysis of the NRPS systems responsible for the lipopeptide production. (Dubern, 2008) However, Muangkaew et al. showed that the structures of putisolvin I and II as published contained errors, and that they should be revised. (Muangkaew, 2024) More so, the structures of putisolvin I and II were found to be identical to those of putisolvin III and IV, respectively.

Putisolvins are CLiPs which possess a peptide moiety of 12 amino acids, whereby cyclization occurs between the side chain of Ser9 and the C-terminal carbonyl. The structure of putisolvin I is shown to be C6 : 0 – L-Leu –D-Glu – D-Leu – L-Leu – D-Gln – D-Ser – D-Val – D-Leu – D-Ser – L-Leu – L-Val – L-Ser. (Muangkaew, 2024) Remarkably, the fatty acid chain lacks the 3-hydroxyl functionality, typifying most Pseudomonas CLiPs. The minor compound produced, putisolvin II, has 14 Da more in mass due to the replacement of a valine to isoleucine on the 11th amino acid position.

Chemical structure of putisolvin I
Schematic representation of putisolvin I-II sequences

Putisolvin II is isobaric to cocoyamide/gacamide, as both compounds have C66H115N13O19 as chemical formula. Hence, they cannot be discriminated based on mass spectrometry alone. Despite the identical chemical formula and molecular mass, both types of lipopeptides are structurally highly distinct, as they differ in amino acid sequence (both identity and length), size of the macrocycle and their fatty acid moiety.

Biological activity

For putisolvins produced by P. putida PCL1445, no antibacterial effects were observed against Pseudomonas fluorescens and Pseudomonas aeruginosa. (Kuiper et al., 2004)

At concentrations of 20–25 mg mL-1, putisolvin immobilizes zoospores from different oomycetes and cause rapid lysis of entire zoospore populations . (Kruijt, 2009; Tran, 2008) Similarly, in vitro assays showed that putisolvin I was able to lyse a zoospore suspension of Phytophthora capsici at concentrations above 10 µM. Furthermore, an in planta assay demonstrated that a 50 μM treatment of putisolvin on a zoospore suspension of Phytophthora capsici led to a significant reduction in Phytophthora blight disease in cucumber plants. (Sheng, 2024)

Putisolvins produced by P. putida inhibited biofilm formation by P. aeruginosa PA14 and P. fluorescens WCS365 (Kuiper et al., 2004).

NMR fingerprint data

It was demonstrated that the planar structure and stereochemistry of CLiPs can be assessed by simple comparison to a reference. (De Roo, 2022; Muangkaew, 2024) More specifically, by matching NMR spectra of a CLiP from a newly isolated bacterial source with those of existing (reference) CLiPs, one can determine whether they are identical or not. A detailed explanation on what NMR fingerprint matching is, and how to use it, can be found here.

Below, we provide the reference NMR data of putisolvin I and II. This data is recorded in DMF-d7 at 298K (25°C), and can be used to asses similarities of newly isolated CLiPs to putisolvin I and II.

References

Bernat, et al. “Characterization of extracellular biosurfactants expressed by a Pseudomonas putida strain isolated from the interior of healthy roots from Sida hermaphrodita grown in a heavy metal contaminated soil.” Current Microbiology76, 11 (2019): https://dx.doi.org/10.1007/s00284-019-01757-x.

De Roo, et al. “An Nuclear Magnetic Resonance Fingerprint Matching Approach for the Identification and Structural Re-Evaluation of Pseudomonas Lipopeptides.” Microbiology Spectrum10, 4 (2022): https://dx.doi.org/doi:10.1128/spectrum.01261-22.

Dubern, et al. “Genetic and functional characterization of the gene cluster directing the biosynthesis of putisolvin I and II in Pseudomonas putida strain PCL1445.” Microbiology (Reading)154, Pt 7 (2008): https://dx.doi.org/10.1099/mic.0.2008/016444-0.

Kruijt, et al. “Functional, genetic and chemical characterization of biosurfactants produced by plant growth-promoting Pseudomonas putida 267.” Journal of Applied Microbiology107, 2 (2009): https://dx.doi.org/10.1111/j.1365-2672.2009.04244.x.

Kuiper, et al. “Characterization of two Pseudomonas putida lipopeptide biosurfactants, putisolvin I and II, which inhibit biofilm formation and break down existing biofilms.” Molecular Microbiology51, 1 (2003): https://dx.doi.org/10.1046/j.1365-2958.2003.03751.x.

Muangkaew, et al. “Breaking Cycles: Saponification-Enhanced NMR Fingerprint Matching for the Identification and Stereochemical Evaluation of Cyclic Lipodepsipeptides from Natural Sources.” Chemistry – A European Journal (2024): https://dx.doi.org/https://doi.org/10.1002/chem.202400667.

Sheng, et al. “The biocontrol roles of cyclic lipopeptide putisolvin produced from Pseudomonas capeferrum HN2-3 on the Phytophthora blight disease in cucumbers.” Journal of Plant Diseases and Protection 131, 2 (2024): https://dx.doi.org/10.1007/s41348-024-00874-5.

 

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