Selection of antifungal protein-producing molds from dry-cured meat products

Acosta, Raquel; Rodrı́guez-Martı́n, Andrea; Martı́n, Alberto; Núñez, Félix; Asensio, Miguel A.

doi:10.1016/j.ijfoodmicro.2009.07.020

Cited by 45 publications

(29 citation statements)

References 27 publications

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“…Mycotoxin production was tested after each mold strain was three‐point inoculated per plate on malt extract agar and incubated at 25°C for 21 days. The content of three petri dishes was extracted according to the method described by Sosa et al (), and the extracts were analyzed by chromatographic separation according to Acosta, Rodríguez‐Martín, Martín, Núñez, and Asensio (), using an Agilent 1100 Series chromatograph (Agilent Technologies Inc., Palo Alto, CA, USA) equipped with a thermostated C18 column (Phenomenex, 150 × 4.6 mm, 5 µm) at 40ºC connected to a DAD detector. The mobile phases were composed of deionized water: methanol 55:45 (v/v) (A) and 20:80 (v/v) (B).…”

Section: Methodsmentioning

confidence: 99%

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Use of efficient drying methods to improve the safety and quality of dried fig

Villalobos

Serradilla

Martı́n

et al. 2018

J Food Process Preserv

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This study aimed to evaluate the influence of preselect artificial drying systems for figs on the safety and sensory quality of this product throughout shelf‐life. Fresh figs were dried by different artificial methods and compared to traditional sun drying as the control. The microbial counts were monitored after drying and throughout storage. The fungal population was isolated and identified, and their capacity to produce mycotoxins was evaluated. Quality parameters such as moisture, weight loss, firmness, total soluble solids, titratable acidity, and sensory quality were also assessed monthly. Ultrasound treatment combined with an osmotic solution presented the lowest microbial counts, in contrast with the traditional dried. No relevant differences were found in fungal species between drying systems, being strains of the genus Penicillium the primary mycotoxin producers (penicillic acid, griseofulvin, and roquefortine). Ultrasound treatment was effective for fig drying, providing a stable, safe and good quality product, increasing its potential storability. Practical applications The safety risk in figs naturally dried, such us the presence of mycotoxins, is a big concern. Likewise, the quality of the fig dried by this traditional method is very heterogenic, being highly conditioned by weather conditions. Until now, there is scarce information about alternative methods to natural drying of this appreciate product. In our study, we approach several drying strategies evaluating safety and quality aspects. These findings should be taken account in the development of specific technologies for the dried fig production.

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Section: Methodsmentioning

confidence: 99%

“…The volume of sample injected was 10 µl. The presence in the extracts of more than 20 mycotoxins (including aflatoxins, ochratoxin A, patulin, citrinin, etc.,) was monitored according to the retention times and UV‐Vis spectra of the metabolites compared to standards provided by Sigma (Sigma Chemical, St. Louis, USA) (Acosta et al, ).…”

Section: Methodsmentioning

confidence: 99%

Use of efficient drying methods to improve the safety and quality of dried fig

Villalobos

Serradilla

Martı́n

et al. 2018

J Food Process Preserv

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“…The cellfree medium was applied to an ÄKTA FPLC with a cationic exchange column HiTrap SP HP (Amersham Biosciences, Uppsala, Sweden), equipped with a UV detector at 214 nm. The resulting fraction containing PgAFP was then chromatographed on a HiLoad 26/60 Superdex 75 column for FPLC (Amersham Biosciences, Uppsala, Sweden) as previously described (Acosta et al 2009) to obtain an extract of the purified protein. The extract containing the purified PgAFP from several batches were pooled in a stock solution.…”

Section: Strainsmentioning

confidence: 99%

Impact of the antifungal protein PgAFP from Penicillium chrysogenum on the protein profile in Aspergillus flavus

Delgado

Owens

Doyle

et al. 2015

Appl Microbiol Biotechnol

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Antifungal proteins produced by molds are generally small, highly basic, and cysteine-rich. The best known effects of these proteins include morphological changes, metabolic inactivation, and membrane perturbation on sensitive fungi. Reactive oxygen species (ROS) generation leads to apoptosis, with G -protein playing a key role in transduction of cell death signals. The antifungal protein PgAFP from Penicillium chrysogenum inhibits growth of some toxigenic molds. Here we analyzed the effect of the antifungal protein PgAFP on the growth of Aspergillus flavus. For this, comparative proteomic analysis was used to identify the whole protein profile and protein change in abundance after PgAFP treatment. PgAFP provoked metabolic changes related to reduced energy metabolism, cell wall integrity alteration, and increased stress response due to higher levels of ROS. The observed changes in protein abundance, favoring a higher glutathione concentration as well as the increased abundance in heat shock proteins, do not seem to be enough to avoid necrosis. The decreased chitin deposition observed in PgAFP-treated A. flavus is attributed to a lower relative quantity of Rho1. The reduced relative abundance of a β subunit of G -protein seems to be the underlying reason for modulation of apoptosis in PgAFP-treated A. flavus hyphae. We propose Rho1 and G -protein subunit β CpcB to be the main factors in the mode of action of PgAFP in A. flavus. Additionally, enzymes essential for the biosynthesis of aflatoxin were no longer detectable in A. flavus hyphae at 24 h, following treatment with PgAFP. This presents a promising effect of PgAFP, which may prevent A. flavus from producing mycotoxins. However, the impact of PgAFP on actual aflatoxin production requires further study.

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“…PgAFP was obtained from P. chrysogenum CECT 20922 grown in potato dextrose broth (PDB, Scharlab, Barcelona, Spain) pH 4.5, at 25°C for 21 days, as described previously (Acosta et al 2009). To get cell-free medium, mycelium was removed by filtering through Miracloth (Calbiochem, Darmstadt, Germany) and the culture medium was filtered through a nitrocellulose 0.22 μm pore size (Sartorius, Goettingen, Germany).…”

Section: Purification Of Pgafpmentioning

confidence: 99%

Increased chitin biosynthesis contributes to the resistance of Penicillium polonicum against the antifungal protein PgAFP

Delgado

Owens

Doyle

et al. 2015

Appl Microbiol Biotechnol

Self Cite

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Antifungal proteins from molds have been proposed as a valuable tool against unwanted molds, but the resistance of some fungi limits their use. Resistance to antimicrobial peptides has been suggested to be due to lack of interaction with the mold or to a successful response. The antifungal protein PgAFP produced by Penicillium chrysogenum inhibits the growth of various ascomycetes, but not Penicillium polonicum. To study the basis for resistance to this antifungal protein, localization of PgAFP and metabolic, structural, and morphological changes were investigated in P. polonicum. PgAFP bound the outer layer of P. polonicum but not regenerated chitin, suggesting an interaction with specific molecules. Comparative two-dimensional gel electrophoresis (2D-PAGE) and comparative quantitative proteomics revealed changes in the relative abundance of several proteins from ribosome, spliceosome, metabolic, and biosynthesis of secondary metabolite pathways. The proteome changes and an altered permeability reveal an active reaction of P. polonicum to PgAFP. The successful response of the resistant mold seems to be based on the higher abundance of protein Rho GTPase Rho1 that would lead to the increased chitin deposition via cell wall integrity (CWI) signaling pathway. Thus, combined treatment with chitinases could provide a complementary means to combat resistance to antifungal proteins.

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Selection of antifungal protein-producing molds from dry-cured meat products

Cited by 45 publications

References 27 publications

Use of efficient drying methods to improve the safety and quality of dried fig

Use of efficient drying methods to improve the safety and quality of dried fig

Impact of the antifungal protein PgAFP from Penicillium chrysogenum on the protein profile in Aspergillus flavus

Increased chitin biosynthesis contributes to the resistance of Penicillium polonicum against the antifungal protein PgAFP

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