ABSTRACT

Metaproteomics enables the large-scale characterization of microbial community proteins, offering crucial insights into their taxonomic composition, functional activities, and interactions within their environments. By directly analyzing proteins, metaproteomics offers insights into community phenotypes and the roles individual members play in diverse ecosystems. Although database-dependent search engines are commonly used for peptide identification, they rely on pre-existing protein databases, which can be limiting for complex, poorly characterized microbiomes. De novo sequencing presents a promising alternative, which derives peptide sequences directly from mass spectra without requiring a database. Over time, this approach has evolved from manual annotation to advanced graph-based, tag-based, and deep learning-based methods, significantly improving the accuracy of peptide identification. This Viewpoint explores the evolution, advantages, limitations, and future opportunities of de novo sequencing in metaproteomics. We highlight recent technological advancements that have improved its potential for detecting unsequenced species and for providing deeper functional insights into microbial communities.

PMID:39888246 | DOI:10.1002/pmic.202400321