Showing papers on "Penicillium griseofulvum published in 2008"

Low occurrence of patulin- and citrinin-producing species isolated from grapes.

Abstract: Aims: To assess the ability of fungi isolated from grapes to produce patulin and citrinin. Methods and Results: A total of 446 Aspergillus isolates belonging to 20 species and 101 Penicillium isolates were inoculated in Czapek yeast extract agar and yeast extract sucrose agar and incubated for 7 days at 25� C. Extracts were analysed for patulin and citrinin by thin-layer chromatography. None of the isolates of Aspergillus spp. produced either patulin or citrinin. Patulin was produced by three isolates of Penicillium expansum and two of Penicillium griseofulvum. Citrinin was produced by five isolates of P. expansum, two of Penicillium citrinum and one of Penicillium verrucosum. Conclusions: Our results show that the Aspergillus and Penicillium species commonly isolated from grapes are not a source of the mycotoxins, patulin and citrinin. Significance and Impact of the Study: The possibility of co-occurrence of patulin and citrinin with ochratoxin A in grapes and grape products remain low, owing to the low frequency of isolation of potentially producing species.

Structure-based mutagenesis of Penicillium griseofulvum xylanase using computational design

Abstract: Penicillium griseofulvum xylanase (PgXynA) belongs to family 11 glycoside hydrolase. It exhibits unique amino acid features but its three-dimensional structure is not known. Based upon the X-ray structure of Penicillium funiculosum xylanase (PfXynC), we generated a three-dimensional model of PgXynA by homology modeling. The native structure of PgXynA displayed the overall beta-jelly roll folding common to family 11 xylanases with two large beta-pleated sheets and a single alpha-helix that form a structure resembling a partially closed right hand. Although many features of PgXynA were very similar to previously described enzymes from this family, crucial differences were observed in the loop forming the "thumb" and at the edge of the binding cleft. The robustness of the xylanase was challenged by extensive in silico-based mutagenesis analysis targeting mutations retaining stereochemical and energetical control of the protein folding. On the basis of structural alignments, modeled three-dimensional structure, in silico mutations and docking analysis, we targeted several positions for the replacement of amino acids by site-directed mutagenesis to change substrate and inhibitor specificity, alter pH profile and improve overall catalytic activity. We demonstrated the crucial role played by Ser44(PgXynA) and Ser129(PgXynA), two residues unique to PgXynA, in conferring distinct specificity to P. griseofulvum xylanase. We showed that the pH optimum of PgXynA could be shifted by -1 to +0.5 units by mutating Ser44(PgXynA) to Asp and Asn, respectively. The S44D and S44N mutants showed only slight alteration in K(m) and V(max) whereas a S44A mutant lost both pH-dependence profile and activity. We were able to produce PgXynA S129G mutants with acquired sensitivity to the Xylanase Inhibitor Protein, XIP-I. The replacement of Gln121(PgXynA), located at the start of the thumb, into an Arg residue resulted in an enzyme that possessed a higher catalytic activity.

A gene having sequence homology to isoamyl alcohol oxidase is transcribed during patulin production in Penicillium griseofulvum.

Abstract: The genes for the patulin biosynthetic pathway are most likely arranged in a cluster, as is often the case for other mycotoxins. With this in mind, GeneWalking has been performed to identify genes both upstream and downstream of the isoepoxydon dehydrogenase (idh) gene. A gene present in Penicillium griseofulvum NRRL 2159A had high sequence homology to the isoamyl alcohol oxidase (iao) gene and was detected downstream of the idh gene and in the same orientation. By virtue of the presence of a signal peptide sequence, the newly identified gene coded for a secreted protein with an FAD-binding domain and potential for N-glycosylation. An open reading frame consisted of 1946 nucleotides, containing four putative introns and encoding a 22 amino acid signal peptide. The 571 amino acid mature protein contained nine cysteine residues and had 11 potential N-linked glycosylation sites. Searches using GenBank indicated that Aspergillus terreus, A. oryzae, A. fumigatus, and Gibberella zeae contain genes coding for a putative isoamyl alcohol oxidase. When the translated query was compared with the translated database, the highest scores were seen with A. clavatus (E value of 0.00), A. fumigatus (E value of 8e−142), and A. oryzae and A. terreus (each having an E value of 2e−141). Reverse transcription-polymerase chain reaction analysis confirmed that the iao gene was transcribed. The amplified products were sequenced for confirmation of their identities. This is the first report of an isoamyl alcohol oxidase gene in a species of the genus Penicillium.

Inhibition of diacylglycerol acyltransferase by phenylpyropenes produced by Penicillium griseofulvum F1959.

Abstract: Phenylpyropenes A, B, and C, isolated from Penicillium griseofulvum F1959, inhibited DGAT in rat liver microsomes with IC50 values of 787+/-16, 217+/-02, and 1104+/-02 mM, respectively In addition, a kinetic analysis using a Lineweaver-Burk plot revealed that phenylpyropene C was a noncompetitive inhibitor of DGAT The apparent Michaelis constant (Km) value and inhibition constant (Ki) value were calculated to be 8 mM and 104 mM, respectively Moreover, phenylpyropene C inhibited triglyceride formation in HepG2 cells

Effect of gamma irradiation on the growth and patulin production of Penicillium griseofulvum in an apple model system

Abstract: The effect of gamma irradiation on the prevention of breeding a patulin-producing mold and reducing patulin content was evaluated in an apple model system. Penicillium griseofulvum, a patulin-producing standard mold strain was artificially inoculated into apples and a gamma irradiation was performed. The D10-values of the conidia of P. griseofulvum in an aqueous suspension and the apple model system were calculated at 0.28 and 0.48 kGy, respectively. The viable cell counts of the inoculated conidia in the apples showed 2 decimal point reductions at a dose of 1 kGy. Breeding and growth of the survived conidia was prevented during 10 weeks of post-irradiation storage period, especially at 4℃. The concentration of patulin in the non-irradiated apples was gradually increased and reached about 950 ppm at 25℃ and 410 ppm at 4℃, but the production of patulin was not observed during storage after 1 kGy of gamma irradiation.

Penicillium griseofulvum F1959, high-production strain of pyripyropene a, specific inhibitor of acyl-CoA: cholesterol acyltransferase 2.

Abstract: Acyl-coenzyme A: cholesterol acyltransferase (ACAT) catalyzes cholesterol esterification and plays an important role in the intestinal absorption of cholesterol, hepatic production of lipoproteins, and accumulation of cholesteryl ester within cells. During the course of screening to find ACAT inhibitors from microbial sources, the present authors isolated pyripyropene A from Penicillium griseofulvum F1959. Pyripyropene A, an ACAT2-specific inhibitor, has already been produced from Aspergillus fumigatus. Yet, Aspergillus fumigatus is a pathogen and only produces a limited amount of pyripyropene A, making the isolation of pyripyropene A troublesome. In contrast, Penicillium griseofulvum F1959 was found to produce approximately 28 times more pyripyropene A than Aspergillus fumigatus, plus this report also describes the ideal conditions for the production of pyripyropene A by Penicillium griseofulvum F1959 and its subsequent purification.