In vivo thrombin activity in the diatom Phaeodactylum tricornutum: Biotechnological insights

Messaabi, A., Merindol, N., Bohnenblust, L., Fantino, E., Meddeb-Mouelhi, F. et Desgagné-Penix, I. (2024). In vivo thrombin activity in the diatom Phaeodactylum tricornutum: Biotechnological insights. Applied Microbiology and Biotechnology, 108 (1). Article 481. ISSN 0175-7598 1432-0614 DOI 10.1007/s00253-024-13322-z

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Résumé

Abstract

Diatoms are responsible for 20% of global carbon dioxide fixation and have significant potential in various biotechnological and industrial applications. Recently, the pennate diatom Phaeodactylum tricornutum has emerged as a prominent platform organism for metabolic engineering and synthetic biology. The availability of its genome sequence has facilitated the development of new bioengineering tools. In this study, we used in silico analyses to identify sequences potentially encoding thrombin-like proteins, which are involved in recognizing and cleaving the thrombin sequence LVPRGS in P. tricornutum. Protein structure prediction and docking studies indicated a similar active site and ligand positioning compared to characterized human and bovine thrombin. The evidence and efficiency of the cleavage were determined in vivo using two fusion-protein constructs that included YFP to measure expression, protein accumulation, and cleavage. Western blot analysis revealed 50–100% cleavage between YFP and N-terminal fusion proteins. Our findings suggest the existence of a novel thrombin-like protease in P. tricornutum. This study advances the application of diatoms for the synthesis and production of complex proteins and enhances our understanding of the functional role of these putative thrombin sequences in diatom physiology.

Key points:
• Protein structure predictions reveal thrombin-like active sites in P. tricornutum.
• Validated cleavage efficiency of thrombin-like protease on fusion proteins in vivo.
• Study advances bioengineering tools for diatom-based biotechnological applications.

Type de document:	Article
Mots-clés libres:	Cleavage peptide Bioengineering Synthetic biology Microalgae Thrombin-like protease YFP (yellow fluorescent protein)
Date de dépôt:	22 avr. 2025 20:23
Dernière modification:	22 avr. 2025 20:23
Version du document déposé:	Version officielle de l'éditeur
URI:	https://depot-e.uqtr.ca/id/eprint/11874

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