# Biophysical Characterization of a Carotenoprotein from Marine Sponge Tedania ignis Reveals Pigment-Dependent Stability and Antibiotic Interactions

## TL;DR

A blue carotenoprotein from a marine sponge is shown to stabilize the protein structure and enhance antibiotic effects.

## Contribution

The study provides the first comprehensive biophysical characterization of a carotenoprotein from the marine sponge Tedania ignis.

## Key findings

- Carotenoid binding is critical for the structural stability of Ti-CP.
- Ti-CP interacts with multiple antibiotics and enhances their activity against bacteria.
- Both Ti-CP and its apoprotein reduce bacterial viability and biofilm formation.

## Abstract

Carotenoproteins from marine sponges represent an underexplored class of pigment–protein complexes with distinctive structural and functional properties. Here, we report the isolation and biophysical characterization of a blue carotenoprotein from the sponge Tedania ignis, termed Ti-CP. The protein was purified and shown to consist of two closely related isoforms with molecular masses of approximately 27–29 kDa. Reverse-phase chromatography enabled separation of the apoprotein (ApoTi-CP) and its associated carotenoids, which were identified as oxygenated carotenoids consistent with astaxanthin and mytiloxanthin. Circular dichroism analysis revealed that both Ti-CP and ApoTi-CP are dominated by β-sheet secondary structure and display highly similar conformational profiles. In contrast, dynamic light scattering demonstrated that carotenoid binding is critical for protein stability, as the native form exhibited a compact and monodisperse organization, whereas ApoTi-CP showed pronounced aggregation. Isothermal titration calorimetry revealed that Ti-CP, but not ApoTi-CP, interacts with tetracycline, oxacillin, and streptomycin, indicating that pigment-mediated stabilization modulates ligand binding. Both Ti-CP and ApoTi-CP reduced bacterial viability and biofilm formation in a strain-dependent manner and enhanced antibiotic activity, including synergistic effects against resistant bacteria. Together, these results provide a comprehensive description of a previously uncharacterized sponge carotenoprotein and highlight the dual role of carotenoids in structural stabilization and antimicrobial modulation, reinforcing the biotechnological relevance of marine pigment–protein complexes.