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Published on: January 21, 2014

Egg Hatching, Peptide Pheromones, and Endoproteinases in Barnacles.

Desa Bolger¹, Joshua Osterberg¹, Beatriz Orihuela¹

¹Duke University Marine Laboratory, Nicholas School of the Environment, 135 Duke Marine Lab Road, Beaufort, NC 28516, USA.

International Journal of Molecular Sciences

|December 11, 2025

Summary

Barnacles use enzymes to clean eggs and release larvae, acting as pheromones. This study identifies key barnacle endoproteinases, crucial for understanding reproductive processes and microbiome interactions.

Keywords:

barnacles egg brooding egg hatching endoproteinase exoproteinases microbiomes proteomics

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Area of Science:

Marine Biology
Biochemistry
Proteomics

Background:

The striped barnacle (Amphibalanus amphitrite) is a simultaneous hermaphrodite crustacean that broods eggs.
Barnacles use enzymatic processes within the mantle to clean eggs during incubation, preventing biofouling.
Enzyme products function as pheromones, regulating egg hatching and larval release, with known pheromones being peptides from trypsin activity.

Purpose of the Study:

To characterize barnacle brooding endoproteinases using proteomic analysis.
To identify specific enzyme sequences and cleavage sites involved in egg hatching and larval release.
To compare enzyme activity and patterns between barnacle samples and seawater.

Main Methods:

Proteomic analysis of peptides generated from pure protein hydrolysis.
High-resolution LC electrospray MS/MS for peptide identification.
Characterization of endoproteinases based on C-terminus amino acids and enzyme activity.

Main Results:

Enzyme activity was significantly higher (2.22–2.79 times) in barnacle and microbiome samples compared to seawater.
Distinct enzyme patterns were observed: barnacle samples showed higher proline- and asparagine-cutting enzymes, while seawater had more elastase.
At least 13 endoproteinases were identified, primarily serine proteases, with complete sequence identification around cleavage sites.

Conclusions:

The study successfully characterized barnacle endoproteinases involved in reproduction.
The findings highlight the role of the barnacle microbiome in enzyme production.
The proteomic approach offers a powerful method for detailed endoproteinase analysis in various microbiome assemblages.