title: Manuscript title: antifungal proteins from moulds: analytical tools and potential application to dry-ripened foods.
abstract: Moulds growing on the surface of dry-ripened foods contribute to their sensory qualities, but some of them are able to produce mycotoxins that pose a hazard to consumers. Small cysteine-rich antifungal proteins (AFPs) from moulds are highly stable to pH and proteolysis and exhibit a broad inhibition spectrum against filamentous fungi, providing new chances to control hazardous moulds in fermented foods. The analytical tools for characterizing the cellular targets and affected pathways are reviewed. Strategies currently employed to study these mechanisms of action include 'omics' approaches that have come to the forefront in recent years, developing in tandem with genome sequencing of relevant organisms. These techniques contribute to a better understanding of the response of moulds against AFPs, allowing the design of complementary strategies to maximize or overcome the limitations of using AFPs on foods. AFPs alter chitin biosynthesis, and some fungi react inducing cell wall integrity (CWI) pathway. However, moulds able to increase chitin content at the cell wall by increasing proteins in either CWI or calmodulin-calcineurin signalling pathways will resist AFPs. Similarly, AFPs increase the intracellular levels of reactive oxygen species (ROS), and moulds increasing G-protein complex β subunit CpcB and/or enzymes to efficiently produce glutathione may evade apoptosis. Unknown aspects that need to be addressed include the interaction with mycotoxin production by less sensitive toxigenic moulds. However, significant steps have been taken to encourage the use of AFPs in intermediate-moisture foods, particularly for mould-ripened cheese and meat products.
authors.affiliation: Food Hygiene and Safety, Institute of Meat Products, University of Extremadura, Cáceres, Spain.
authors.forename: Josué
authors.initials: J
authors.lastname: Delgado
authors.affiliation: Department of Biology, Maynooth University, Maynooth, Co. Kildare, Ireland.
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Owens
authors.affiliation: Department of Biology, Maynooth University, Maynooth, Co. Kildare, Ireland.
authors.forename: Sean
authors.initials: S
authors.lastname: Doyle
authors.affiliation: Food Hygiene and Safety, Institute of Meat Products, University of Extremadura, Cáceres, Spain. masensio@unex.es.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Asensio
authors.affiliation: Food Hygiene and Safety, Institute of Meat Products, University of Extremadura, Cáceres, Spain.
authors.forename: Félix
authors.initials: F
authors.lastname: Núñez
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology*; D002686:Chitin / Q000096:biosynthesis; D005506:Food Contamination / Q000517:prevention & control*; D005516:Food Microbiology; D059022:Food Safety; D005656:Fungal Proteins / Q000494:pharmacology; D005658:Fungi / Q000187:drug effects / Q000378:metabolism*; D009183:Mycotoxins / Q000096:biosynthesis; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D023181:Antimicrobial Cationic Peptides; D005656:Fungal Proteins; D009183:Mycotoxins; D017382:Reactive Oxygen Species; C097609:antifungal protein, Aspergillus; D002686:Chitin
keywords: Antifungal proteins; Food safety; Proteomics; Toxigenic moulds