Characterization of the proteolytic activity of starter cultures of Penicillium roqueforti for production of blue veined cheeses

Larsen, M

doi:10.1016/s0168-1605(98)00114-7

Cited by 61 publications

(23 citation statements)

References 16 publications

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“…However, identification programs are of value for the selection of new strains to be used as starters for the production of blue cheeses (Larsen, Kristiansen, & Hansen, 1998;Dupont et al, 1999;Geisen et al, 2001). The same species found in the present study, and their distribution between the surface and interior samples, have been reported by other authors who have analysed Cabrales cheese (Nu´n˜ez, Medina, Gaya, Dias-Amado, 1981).…”

Section: Discussionsupporting

confidence: 88%

Morphotypic and molecular identification of filamentous fungi from Spanish blue-veined Cabrales cheese, and typing of Penicillium roqueforti and Geotrichum candidum isolates

Flórez

Álvarez‐Martín

López-Dı́az

et al. 2007

International Dairy Journal

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Section: Discussionsupporting

confidence: 88%

Morphotypic and molecular identification of filamentous fungi from Spanish blue-veined Cabrales cheese, and typing of Penicillium roqueforti and Geotrichum candidum isolates

Flórez

Álvarez‐Martín

López-Dı́az

et al. 2007

International Dairy Journal

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“…The proteolytic activity of B. clausii supernatants (OC-SN) was tested preliminarily using agar casein hydrolysis on agar plates containing yeast nitrogen base (YNB) agar medium (Difco Laboratory, Detroit, MI, USA) supplemented with 0.5% casein (35).…”

Section: Methodsmentioning

confidence: 99%

Secreted Compounds of the Probiotic Bacillus clausii Strain O/C Inhibit the Cytotoxic Effects Induced by Clostridium difficile and Bacillus cereus Toxins

Ripert

Racedo

Elie

et al. 2016

Antimicrob Agents Chemother

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Although the use of probiotics based on Bacillus strains to fight off intestinal pathogens and antibiotic-associated diarrhea is widespread, the mechanisms involved in producing their beneficial effects remain unclear. Here, we studied the ability of compounds secreted by the probiotic Bacillus clausii strain O/C to counteract the cytotoxic effects induced by toxins of two pathogens, Clostridium difficile and Bacillus cereus, by evaluating eukaryotic cell viability and expression of selected genes. Coincubation of C. difficile and B. cereus toxic culture supernatants with the B. clausii supernatant completely prevented the damage induced by toxins in Vero and Caco-2 cells. The hemolytic effect of B. cereus was also avoided by the probiotic supernatant. Moreover, in these cells, the expression of rhoB, encoding a Rho GTPase target for C. difficile toxins, was normalized when C. difficile supernatant was pretreated using the B. clausii supernatant. All of the beneficial effects observed with the probiotic were abolished by the serine protease inhibitor phenylmethylsulfonyl fluoride (PMSF). Suspecting the involvement of a secreted protease in this protective effect, a protease was purified from the B. clausii supernatant and identified as a serine protease (M-protease; GenBank accession number Q99405). Experiments on Vero cells demonstrated the antitoxic activity of the purified protease against pathogen supernatants. This is the first report showing the capacity of a protease secreted by probiotic bacteria to inhibit the cytotoxic effects of toxinogenic C. difficile and B. cereus strains. This extracellular compound could be responsible, at least in part, for the protective effects observed for this human probiotic in antibiotic-associated diarrhea.T he gut, one of the biggest surfaces of exchange between the body's internal and external environment, is in permanent contact with antigens. Indeed, the intestinal barrier constitutes an important playground for numerous intestinal pathogen bacteria and toxins.Among them, Clostridium difficile, an anaerobic Gram-positive bacterium, is a major cause of diarrhea, pseudomembranous colitis, and septicemia, and it can lead to death. Hospital stay and use of antibiotics are major risk factors for contracting this disease (1). Over the past decade, the frequency and severity of C. difficile infections have increased markedly due to the emergence of socalled hypervirulent strains that overproduce toxins (2-4). The C. difficile toxins modify target cell proteins to cause disassembly of the actin cytoskeleton and induce severe inflammation (5).Expression of several host genes, mainly those involved in cellular processes, cell-cell interactions, apoptosis, and inflammation, is modified during C. difficile infection (6). These include host genes encoding RhoB (regulating actin cytoskeleton and interactions between cells), ZO-1 (tight-junction protein), and ROCK2 (actin cytoskeleton and signaling protein regulator) (7-9).Equally, many other pathogens, such as Salmonella species, E...

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“…INBCC 300. Larsen et al (1998) described that only high extracellular protease producers, with volumetric values of activity in excess of 70 U/ml at pH 6.0, can be detected by qualitative assays. Our positive control Penicillium sp.…”

mentioning

confidence: 99%

“…The number of transformants is the average of three independent experiments P. roqueforti P20 7.1 ± 1.2 P. camemberti and P. roqueforti were grown in liquid CAC medium (Moralejo et al 2002) to induce proteolytic activity. Aliquots (200-400 ll) were taken at 96 h, and proteolytic activity was measured at pH 6.0 using the azocasein method (Larsen et al 1998). Volumetric activity (U/mL) was defined as by Larsen et al (1998), and normalized for total protein content measured by the Bradford method (Bradford 1976), thus giving specific activity.…”

mentioning

confidence: 99%

“…Aliquots (200-400 ll) were taken at 96 h, and proteolytic activity was measured at pH 6.0 using the azocasein method (Larsen et al 1998). Volumetric activity (U/mL) was defined as by Larsen et al (1998), and normalized for total protein content measured by the Bradford method (Bradford 1976), thus giving specific activity. Values are the average of three independent experiments…”

mentioning

confidence: 99%

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Evaluation of properties of several cheese-ripening fungi for potential biotechnological applications

et al. 2010

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Strains of Penicillium camemberti and Penicillium roqueforti were tested for properties that could be important for future biotechnological applications of these fungi. Penicillium camemberti CECT 2267 and P. roqueforti NRRL 849 showed the most promising performances in terms of growth, protoplast production, and protoplast regeneration abilities. Transformation of these strains with a plasmid containing gene encoding phleomycin resistance showed that they also have a high transformation frequency. In addition, both strains showed low extracellular proteolytic activity. Thus, these strains have all the characteristics to make them suitable for future genetic improvement, recombinant protein production, and other potential biotechnological applications.This work was supported by FONDECYT Grant No. 11060003 and DICYT-USACH

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Characterization of the proteolytic activity of starter cultures of Penicillium roqueforti for production of blue veined cheeses

Cited by 61 publications

References 16 publications

Morphotypic and molecular identification of filamentous fungi from Spanish blue-veined Cabrales cheese, and typing of Penicillium roqueforti and Geotrichum candidum isolates

Morphotypic and molecular identification of filamentous fungi from Spanish blue-veined Cabrales cheese, and typing of Penicillium roqueforti and Geotrichum candidum isolates

Secreted Compounds of the Probiotic Bacillus clausii Strain O/C Inhibit the Cytotoxic Effects Induced by Clostridium difficile and Bacillus cereus Toxins

Evaluation of properties of several cheese-ripening fungi for potential biotechnological applications

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