Transcriptomics-Guided Biodiscovery of Peroxidases from Macroalgae

Peroxidases are versatile enzymes with applications in laboratory analytics and industrial processes. Despite their relevance, many applications rely on a few well-characterized terrestrial enzymes, particularly horseradish peroxidase (HRP), whose complex post-translational modifications hinder cost-efficient recombinant production. Marine macroalgae, due to their phylogenetic diversity and ecological adaptations, represent a promising and largely untapped source of novel biocatalysts. The aim of this work was the transcriptomics-guided identification and characterization of novel enzymes, particularly peroxidases, from macroalgae. Adaptable RNA extraction protocols were developed for green, red, and brown algae, as well as seagrass and cyanobacteria. Using a CTAB-based lysis buffer and multistep purification, intact RNA was isolated from most of the 22 tested samples. These extracts and additional datasets were used to assemble and annotate ten transcriptomes comprising over 19,000 contigs. Approximately 70 candidate vanadium-dependent haloperoxidase (vHPO) genes were identified. They showed conserved catalytic motifs but high sequence diversity. Transcriptional analysis in Saccharina latissima under stress conditions indicated upregulation, particularly under oxidative stress, suggesting a role in stress response. In addition, a cytochrome c peroxidase from Bryopsis plumosa (BpCcP) was identified and recombinantly expressed in Escherichia coli as a fusion with a single-domain antibody (VHH). The fusion protein was successfully applied as a reporter enzyme in an ELISA, demonstrating both target binding and a detectable signal. Overall, this work establishes a transcriptome-based approach for enzyme discovery in macroalgae and highlights their potential as a source of biotechnologically relevant peroxidases. The generated transcriptome data provide a valuable resource for future research.

Level thesis:degree_level

thesis.doctoral

Grantor dc:publisher

Christian-Albrechts-Universität zu Kiel

Year

2025

Author dc:creator

• Jensen, Timo

Contributors dc:contributor

• Labes, Antje
• Schulz, Rüdiger

• macroalgae
• peroxidases
• vanadium-dependent haloperoxidases
• transcriptomics
• enzyme discovery
• recombinant expression
• cytochrome c peroxidase
• reporter enzyme

Repository record source_url

https://macau.uni-kiel.de/receive/macau_mods_00008153

OAI identifier oai:identifier

oai:macau.uni-kiel.de:macau_mods_00008153