Purification and characterization of a novel antifungal protein secreted by Penicillium chrysogenum from an Arctic sediment

Chen, Zhiteng; Ao, Jingqun; Yang, Wenchuan; Jiao, Liping; Zheng, Tao; Chen, Xinhua

doi:10.1007/s00253-013-4800-6

Cited by 39 publications

(41 citation statements)

References 31 publications

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“…As previously reported, P. italicum (Abidi et al, 2014), P. chrysogenum (Chen et al, 2013), Botrytis cinerea (Abidi et al, 2011), Graphium putredinis , and Trichoderma harzianum (Savitha et al, 2011) produce proteases with an optimum temperature of 50°C, whereas the serine protease from P. waksmanii (Graminho et al, 2013) exhibits maximum activity at 35°C. A metalloprotease isolated from Thermoascus aurantiacus was found to have high proteolytic activity at 75°C (Merheb-Dini et al, 2009), and that from Termitomyces clypeatus was found to have high proteolytic activity at 45°C (Majumder et al, 2015).…”

Section: Discussionsupporting

confidence: 58%

Analysis of the Specificity and Biochemical Characterization of Metalloproteases Isolated from Eupenicillium javanicum Using Fluorescence Resonance Energy Transfer Peptides

Neto

Oliveira

et al. 2017

Front. Microbiol.

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Enzymes have important features that may facilitate their application in industrial processes and have been used as alternatives to chemical catalysts. In particular, proteases can be isolated from microorganisms, which provide important sources of advantageous enzymes for industrial processes. For example, Eupenicillium javanicum is a filamentous fungus that has been shown to express industrially applicable enzymes and chemical components, such as antifungal compounds. The biotechnological potential of E. javanicum and proteases made us search a novel protease from this microorganism. The macromolecule was isolated, the main biochemical properties was evaluated, and the specificity of the protease subsites was determined. The protease was produced under solid-state bioprocess with wheat bran and isolated by two chromatography steps with yield of 27.5% and 12.4-fold purification. The molecular mass was estimated at 30 kDa. The N-terminal sequence of the first 20 amino acid residues was AVGAGYNASVALALEKALNN. The enzyme presented higher proteolytic activity at pH 6.0 and 60°C. The protease is stable at wide range of pH values and temperatures and in the presence of surfactants. The “primed” side of the catalytic site showed the highest catalytic efficiency of the enzyme isolated from E. javanicum. The S′1 subsite is responsible for catalyzing the protease reaction with substrates with tyrosine in P′1. These findings provide important insights into the biochemical characterization of a highly active protease from E. javanicum and may facilitate the development of industrial processes involving this protease.

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Section: Discussionsupporting

confidence: 58%

Analysis of the Specificity and Biochemical Characterization of Metalloproteases Isolated from Eupenicillium javanicum Using Fluorescence Resonance Energy Transfer Peptides

Neto

Oliveira

et al. 2017

Front. Microbiol.

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“…Some antifungal proteins produced by molds are small, highly basic, and cysteine-rich proteins, such as PAF from Penicillium chrysogenum Q176 (Marx et al 1995), PgAFP from P. chrysogenum RP42C (Rodríguez-Martín et al 2010), PcArctin from P. chrysogenum A096 (Chen et al 2013), AFP from Aspergillus giganteus (Nakaya et al 1990), or NAFP from Neosartorya fischeri . The expression of these proteins is induced in the presence of other fungi and contributes to an ecological advantage (Marx 2004).…”

Section: Introductionmentioning

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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“…Other antifungal proteins produced by ascomycetes are PAF from P. chrysogenum Q176 (Marx et al 1995), PcArctin from P. chrysogenum A096 (Chen et al 2013), BP from Penicillium brevicompactum (Seibold et al 2011), AFP and AFP NN5353 from Aspergillus giganteus (Nakaya et al 1990;Binder et al 2011), Anafp from Aspergillus niger (Gun , AcAFP and AcAMP from Aspergillus clavatus (Skouri-Gargouri and Gargouri 2008; Hajji et al 2010), FPAP from Fusarium polyphialidicum (Galgóczy et al 2013b), and NFAP from Neosartorya fischeri (Kovács et al 2011). Mechanisms of action of antifungal proteins from molds have been described as multifactorial, where membrane permeabilization, changes in actin distribution, chitin biosynthesis inhibition, destabilization of cell wall, Electronic supplementary material The online version of this article (doi:10.1007/s00253-015-7020-4) contains supplementary material, which is available to authorized users.…”

Section: Introductionmentioning

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

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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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Purification and characterization of a novel antifungal protein secreted by Penicillium chrysogenum from an Arctic sediment

Cited by 39 publications

References 31 publications

Analysis of the Specificity and Biochemical Characterization of Metalloproteases Isolated from Eupenicillium javanicum Using Fluorescence Resonance Energy Transfer Peptides

Analysis of the Specificity and Biochemical Characterization of Metalloproteases Isolated from Eupenicillium javanicum Using Fluorescence Resonance Energy Transfer Peptides

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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