# Diving into the chemistry of sponge exo-metabolomes: contribution of brominated specialized metabolites

Morgane Mauduit, Stephane Greff, Thierry Pérez, Charlotte Simmler

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Morgane Mauduit, Stephane Greff, Thierry Pérez, Charlotte Simmler. Diving into the chemistry of sponge exo-metabolomes: contribution of brominated specialized metabolites. Ecole thématique METABIODIVEX, Jun 2022, Aussois, France. 10.5281/zenodo.7254114. hal-03857554

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Submitted on 17 Nov 2022

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<figure><img src="image_1.png" /></figure>

Distributed under a Creative Commons Attribution 4.0 International License
# Diving into the chemistry of sponge exo-metabolomes: contribution of brominated specialized metabolites

Morgane MAUDUIT¹, Stéphane GREFF¹, Thierry PEREZ¹, Charlotte SIMMLER¹

¹ Institut Méditerranéen de Biodiversité et d'Écologie Marine et Continentale, UMR 7263
Endoume Marine Station, rue de la batterie des lions, 13007 Marseille
(e-mail: morgane.mauduit@imbe.fr)

SESAM
project

Chemical Diversity
Chemical Knowledge

<figure><img src="image_2.png" /><figcaption>EXO-METABOLOME</figcaption></figure>

## Context

* Sponges produce **structurally diverse specialized metabolites** known for their biological/pharmacological potentials.¹,² In particular, the Mediterranean sponges *Aplysina cavernicola* and *Agelas oroides* produce **brominated** alkaloids with antimicrobial/anti-biofilm properties.³,⁴

* A few targeted studies, performed on Caribbean species from the same genus, reported that the most abundant brominated alkaloid (e.g. aerothionin, oroidin) were detected in the seawater surrounding the sponges (●●●).⁵,⁶

* Hence, such exo-metabolites (EM) could be collected from the seawater without destroying the sponge biomass.

<figure><img src="image_3.png" /></figure>

What is the proportion (diversity & quant.) of specialized metabolites released by *A. cavernicola* & *A. oroides* in their surroundings?

<figure><img src="image_4.png" /><figcaption>*Aplysina cavernicola* (Vacelet, 1959)</figcaption></figure>

<figure><img src="image_5.png" /><figcaption>Bromotyrosine
spiroisoxazoline alkaloids</figcaption></figure>

<figure><img src="image_6.png" /><figcaption>*Agelas oroides* (Schmidt, 1864)</figcaption></figure>

<figure><img src="image_7.png" /><figcaption>Bromopyrrole alkaloids</figcaption></figure>

<caption>METABOLOME</caption>

## ① Sample preparation

<figure><img src="image_8.png" /></figure>

## ② MS² acquisition & processing

<figure><img src="image_9.png" /><figcaption>LC-HRMS</figcaption></figure>

<figure><img src="image_10.png" /></figure>

<figure><img src="image_11.png" /></figure>

<figure><img src="image_12.png" /></figure>

.CSV

<figure><img src="image_13.png" /></figure>

.mgf

## ③ Identification & Quantification

<figure><img src="image_14.png" /><figcaption>Molecular Network</figcaption></figure>

<figure><img src="image_15.png" /></figure>

## Results

### 1 Exo-metabolites (EM) concentration

<figure><img src="image_16.png" /></figure>

* Comparison of different polymeric resins → Solid phase selection according sponge chemistry

* Variation of the experiment duration

* Metabolite adsorption with/without stress

### 2 Chemical diversity of *A. cavernicola* EM

<figure><img src="image_17.png" /></figure>

Br

<figure><img src="image_18.png" /></figure>

.CSV

Br₂

<figure><img src="image_19.png" /></figure>

Br₄

<figure><img src="image_20.png" /></figure>

<figure><img src="image_21.png" /><figcaption>Analysis of Mzmine extracted features
MS isotopic pattern</figcaption></figure>

Reproducibly
detected

<figure><img src="image_22.png" /><figcaption>Chemical superclass
Sirius score > 100</figcaption></figure>

* 11% of the total detected EM features were found to be **brominated** (structures with 2/4 Br) & could correspond to *A. cavernicola* specialized metabolites.

* 79% of the reproducibly detected EM features belong to the class of lipids and lipid like molecules.

### 3 Identification of brominated exo-metabolites

FBMN : Bromotyrosine
spiroisoxazoline

<figure><img src="image_23.png" /><figcaption>¹H NMR spectrum of aerothionin (CD₃OD, 600 MHz)</figcaption></figure>

<figure><img src="image_24.png" /></figure>

* A total of 6 characteristic **brominated** alkaloids identified by MS and NMR analyses after purification were detected in the seawater, including aerothionin.

* All their MS and NMR data will be shared in open access database.

## Conclusions & Perspectives

* The present experimental and analytical workflow enable us to determine the proportion & identity of brominated exo-metabolites (EM) recovered from the seawater surrounding *A. cavernicola*. The same workflow was implemented with *A. oroides*. Quantitative analysis are in progress. Collectively, those EM are possibly released through the sponge metabolic activities & cellular turnover.⁷

* Such results will serve as a stepping stone to: • guide **in situ** field work to study the release of specialized metabolites by both sponge species

• propose a **valorisation of sponge chemical diversity without collecting their biomass**

## References:

1. Carroll, A. R., et al. Nat. Prod. Rep. (2022) 2. Pawlik, J., et al. Mar. Ecol. Prog. Ser. 127, 183–194 (1995) 3. Peng, J., et al. Alkaloids Chem Biol. 61: 59–262 (2005) 4. Richards, J., et al. Bioorg Med Chem Lett. 18(15):4325-7 (2008) 5. Walker, R. P. et al. Mar Biol 88, 27-32 (1985), 6. Richelle-Maurer, E, et al. Biochemical Systematics and Ecology 31, 1073–1091, (2003) 7. de Goeij, J. M., et al. Science 342, 108–110 (2013)