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We previously identified a novel application for chitinases wherein the <jats:italic>Candida albicans<\/jats:italic> cell wall surface chitinase 3 (Cht3) was shown to have potential in vaccine applications as a subunit antigen against fungal infections. In the present study, this enzyme was investigated further, developing production and purification protocols, enriching our understanding of its properties, and advancing its application potential. Cht3 was heterologously expressed in <jats:italic>Pichia pastoris<\/jats:italic> and a 4\u2010step purification protocol developed and optimised: this involves activated carbon treatment, hydrophobic interaction chromatography, ammonium sulphate precipitation, and gel filtration chromatography. The recombinant enzyme was shown to be mainly <jats:italic>O<\/jats:italic>\u2010glycosylated and to retain the epitopes of the native protein. Functional studies showed it to be highly specific, displaying activity on chitin, chitosan, and chito\u2010oligosaccharides larger than chitotriose only. Furthermore, it was shown to be a stable enzyme, exhibiting activity, and stability over broad pH and temperature ranges. This study represents an important step forward in our understanding of Cht3 and contributes to its development for application.<\/jats:p>","DOI":"10.1002\/biot.202300219","type":"journal-article","created":{"date-parts":[[2023,10,25]],"date-time":"2023-10-25T01:40:29Z","timestamp":1698198029000},"source":"Crossref","is-referenced-by-count":7,"title":["<i>Candida albicans<\/i> chitinase 3 with potential as a vaccine antigen: production, purification, and characterisation"],"prefix":"10.1002","volume":"19","author":[{"given":"Augusto","family":"Costa\u2010Barbosa","sequence":"first","affiliation":[{"name":"Centre of Molecular and Environmental Biology (CBMA)\/Aquatic Research Network (ARNET), Department of Biology University of Minho  Braga Portugal"},{"name":"Institute of Science and Innovation for Bio\u2010Sustainability (IB\u2010S) University of Minho  Braga Portugal"}]},{"given":"Diogo","family":"Ferreira","sequence":"additional","affiliation":[{"name":"Centre of Molecular and 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