Development of Advanced Bioinformatic Profiles to Improve the Detection and Functional Understanding of Fungal Acid Phosphatases

We have retrieved approximately 9,000 protein sequences annotated as fungal acid phosphatase or phytase from the UniProtKB database. After stringent quality filtering, a curated dataset comprising 3,058 high-confidence sequences was assembled. Phylogenetic analysis revealed that these enzymes segregate into eight distinct clades, representing distinct groups of fungal acid phosphatases, purple acid phosphatases, phytases and groups containing both phytases and acid phosphatases annotations. Based on this classification, we have developed three representative protein profiles referred to as Pfr-A-Fungal_phos, Pfr-B-Fungal_phos and Pfr-C-Fungal_phos, each designed to capture the phylogenetic and functional of the enzyme families. The specificity and breadth of these profiles were validated through comprehensive heat-map analyses. When deployed on public protein and metagenomic databases, these profiles demonstrated high sensitivity and specificity, enabling the identification of hundreds of previously uncharacterized fungal proteins. These proteins spanned a broad taxonomic distribution, with notable prevalence in the Ascomycota and Basidiomycota phyla. Collectively, these findings highlight the utility of the newly developed profiles to uncover a novel and taxonomically widespread family of fungal enzymes. This work provided valuable insights into the evolutionary diversity and ecological significance of fungal enzymes involved in phosphorous cycling.