Isolation, Identification, and Characterization of the Algicidal Micromycete Penicillium chrysogenum SR-1.3

Phosphate-solubilizing fungi (PSF) efficiently dissolve insoluble phosphates through the production of organic acids. This study investigates the mechanisms of organic acid secretion by PSF, specifically Penicillium chrysogenum, under tricalcium phosphate (Ca3(PO4)2, Ca-P) and ferric phosphate (FePO4, Fe-P) conditions. Penicillium chrysogenum exhibited higher phosphorus (P) release efficiency from Ca-P (693.6mg/L) than from Fe-P (162.6mg/L). However, Fe-P significantly enhanced oxalic acid (1193.7mg/L) and citric acid (227.7mg/L) production by Penicillium chrysogenum compared with Ca-P (905.7 and 3.5mg/L, respectively). The presence of Fe-P upregulated the expression of genes and activity of enzymes related to the tricarboxylic acid cycle, including pyruvate dehydrogenase and citrate synthase. Additionally, Fe-P upregulated the expression of chitinase and endoglucanase genes, inducing a transformation of Penicillium chrysogenum mycelial morphology from pellet to filamentous. The filamentous morphology exhibited higher efficiency in oxalic acid secretion and P release from Fe-P and Ca-P. Compared with pellet morphology, filamentous morphology enhanced P release capacity by > 40% and > 18% in Ca-P and Fe-P, respectively. This study explored the strategies employed by PSF to improve the dissolution of different insoluble phosphates.

Variation in the carbon to nitrogen ratio in the range 10:1 to 1000:1 had only a small effect on the radial growth rate of four species of fungus on a basic nutrient agar medium. There was no obvious pattern to the differences observed, whether the nitrogen source was organic or inorganic or the medium osmotic potential high or low. The final fresh weight of the colonies was more sensitive to C: N ratio and was depressed by high C: N ratios by as much as 90 % in some treatments. Polyporus versicolor L. ex Fr. was the most efficient accumulator of biomass at C: N of 1000:1, but Penicillium chrysogenum Thorn had the highest mycelial N content. Colony yield did not show the same relationship to external osmotic potential as did the radial growth rate, and the ratio at which the maximum yield at each osmotic potential occurred varied with the different treatments. Turgor potential was reduced by high C:N in the majority of treatments in Polyporus versicolor, Poria vincta (Berk.) Cke. and Phytophthora cinnamomi Rands but not at all in Penicillium chrysogenum. Water content increased with the C: N ratio and decreased with external osmotic potential in all the species except Phytophthora cinnamomi. There was no clear effect of C:N ratio on any of the hyphal solutes measured, that is Na+ , K+ , Cl- , proline or glycerol. Lower concentrations of the organic solutes were present in Phytophthora cinnamomi and Penicillium chrysogenum in these conditions than when KCl and Czapek-Dox with yeast extract medium was used; turgor potential of Penicillium chrysogenum also was 50% lower. Sodium chloride was used to adjust the external osmotic potential and Na+ and Cl- were taken up in increasing amounts as the osmotic potential decreased in all species. Potassium concentrations were similar between species and treatments. The hyphal osmotic potentials, calculated from these solutes alone, were lower (more negative) than the values measured in Poria vincta grown at -1 MPa and Penicillium chrysogenum grown at - 10MPa, but very close to those for Phytophthora cinnamomi grown at - 2.5 MPa.

Over recent decades, a steady increase in the number of allergic diseases has been shown. Current evidence demonstrate a close association between their emergence and exposure to fungal allergens. In this regard, the aim of the present study was to identify frequency and structure of sensitization to the most clinically significant molds in the patients with respiratory allergic diseases. In blood serum of 283 patients with allergic rhinitis and bronchial asthma, we determined total IgE and sIgE to the mold allergens: Penicillium notatum, Cladosporium herbarum, Aspergillus fumigatus and Alternaria alternata by the ImmunoCAP method (Phadia, Sweden). Statistical analysis was carried out by nonparametric methods. The total IgE levels (420 (225.5-641) kU/l) were higher (p 0.05) in patients with sensitization to fungal allergens than in general group (296 (129- 530) kU/l). Multiple sensitization to respiratory allergens was revealed in the patients with allergic rhinitis and bronchial asthma, and sensitization to fungal allergens was associated with increasingly severe manifestations of the disease. In the patients with fungal allergies, sIgE to Alternaria alternata was most often detected (92.5%), with average level of 3.52 (0.635-19.525) kUA/l. Sixteen patients (40%) were sensitized to Aspergillus fumigatus (0.14 (0.06-0.63) kUA/l). In 19 patients (47.5%), we found increased levels of sIgE to Cladosporium herbarum (0.29 (0.045-1.005) kUA/l). Sensitization to Penicillium notatum was detected in 12 patients (30%), the sIgE levels were 0.125 (0.01-0.5) kUA/l. Detection rates in the total group of fungus-allergic patients with respiratory allergies were as follows: Penicillium notatum, 4.2%; Cladosporium herbarum, 6.7%; Aspergillus fumigatus, 5.6%; Alternaria alternata, 13.07%. We found a significant correlation (p 0.05) between the sIgE contents to different fungal allergens. The levels of IgE antibodies to Alternaria alternata correlated with the levels of sIgE to other fungi (Aspergillus fumigates, r = 0.45; Cladosporium herbarum, r = 0.39; Penicillium notatum, r = 0.39). These findings allow us to suggest that sensitization to Alternaria alternata (13.07%) and Cladosporium herbarum (6.7%) is most common among the patients with allergic rhinitis and bronchial asthma, whereas fungal sensitization aggravates clinical course of these diseases. Determination of sIgE to Alternaria alternata can serve as a marker for the presence of potential cross-sensitization to other fungal allergens, i.e., Aspergillus fumigatus, Cladosporium herbarum, and Penicillium notatum.

Ten moulds isolated from municipal waste soil sample were screened for pectinolytic enzyme production when grown on pectin containing (YPSS) solid media. Penicillium chrysogenum was selected based on clearance zones and pectinase enzyme production was carried out in submerged fermentation. Enzyme production by Penicillium chrysogenum was higher at pH 6.5 and a temperature of 35°C using sucrose and ammonium per sulphate as carbon source and nitrogen source, respectively. The maximal activity of P. chrysogenum pectinase was at 50°C, pH 6.5 and was thermostable up to 40°C. MgCl2 and CaCl2 ions had little effect on pectinase activity. Km and Vmax values were 1.0 mg/mL and 85 U/mg protein, respectively and an apparent molecular weight of 31 kDa on SDS-PAGE. Keywords: Penicillium chrysogenum; submerged fermentation, pectinase.

The influence of different growth media and different isolates of Stachybotrys chartarum on monoclonal antibody (Mab) reactivity

The heterologous expression in Aspergillus nidulans of a gene involved in tryptophan biosynthesis from Penicillium chrysogenum is described. With the chimeric plasmid pPC-31, which carries the cloned trpC gene, approximately 10-40 "stable" transformants per microgram of DNA were obtained, with selection for complementation of the mutant allele. This frequency was increased 10-fold by the insertion of the ans1 fragment into the transformation vector. Southern hybridization analysis revealed that transformation occurred as a consequence of the integration of vector sequences into the host chromosome at a variety of sites within the genome.

Role of the carbohydrate moiety of phospholipase B from Penicillium notatum in enzyme activity

Allergy to ingested mycoprotein in a patient with mold spore inhalant allergy

The bioreceptivity, and the consequent biodeterioration of contemporary glass, used by artists worldwide, was studied. The two main objectives were: first, to verify if fungi with some culture media would produce more damages than the same fungi without a nutritional source, and to verify if the two genera of fungi produce the same damage on the same glass. Colourless glass samples with Spruce Pine 87 Batch (SPB-87) composition were inoculated with two distinct fungal species, Penicillium chrysogenum and Aspergillus niger, separately: (i) half with fungal spores (simulating primary bioreceptivity), and (ii) half with fungi in a small portion of culture media (simulating organic matter that can be deposited on exposed glassworks, i.e., secondary bioreceptivity). The alteration of glass surfaces were analysed by Optical Microscopy, SEM-EDS and µ-Raman. The mycelium of Penicillium chrysogenum generated a higher amount of fingerprints, stains and iridescence, whereas Aspergillus niger produced more biopitting and crystals on the glass surface. However, both species damaged the glass to different degrees in 4 and 6 months after the inoculation, producing physico-chemical damage (e.g., iridescence, biopitting), and chemical alterations (e.g., depletion and deposition of elements and crystals). The primary bioreceptivity experiment of glass samples inoculated with Aspergillus niger results in less damage than in the case of secondary bioreceptivity, being almost similar for Penicillium chrysogenum. The new and in-depth understanding of the bioreceptivity and deterioration of post-modern glass art and cultural heritage provided here is of paramount importance for the scientific, conservation and artistic communities—to protect glass cultural materials, or seen by artists as innovative and inspirational ways of creating glass art in the future.

ACUTE RESPIRATORY FAILURE WITH FATAL OUTCOME DUE TO PENICILLIUM CHRYSOGENUM IN A MAN FROM GUATEMALA WITH A SEMINOMA (GERM CELL TUMOR )

Penicillium chrysogenum was immobilized in polyacrylamide gel prepared from 5% acrylamide monomers (85% acrylamide and 15% N,N'-methylene bisacrylamide). Penicillin produced from glucose by the immobilized mycelium was 17% of that produced by washed mycelium. However, the activity of penicillin production of the washed mycelium decreased with repeated use. On the other hand, the activity of the immobilized mycelium increased initially and decreased gradually with repeated use. The rate of oxygen uptake of the immobilized mycelium was about 30% of that of the washed mycelium. The immobilized mycelium required oxygen for the production of penicillin.

The final step in the biosynthesis of beta-lactam antibiotics in Penicillium chrysogenum and Aspergillus nidulans involves removal of the L-alpha-aminoadipyl side chain from isopenicillin N (IPN) and exchange with a nonpolar side chain. The enzyme catalyzing this reaction, acyl-coenzyme A:isopenicillin N acyltransferase (acyltransferase), was purified from P. chrysogenum and A. nidulans. Based on NH2-terminal amino acid sequence information, the acyltransferase gene (penDE) from P. chrysogenum and A. nidulans were cloned. In both organisms, penDE was located immediately downstream from the isopenicillin N synthetase gene (pcbC) and consisted of four exons encoding an enzyme of 357 amino acids (approximately 40 kilodaltons [kDa]). The DNA coding sequences showed approximately 73% identity, while the amino acid sequences were approximately 76% identical. Noncoding DNA regions (including the region between pcbC and penDE) were not conserved. Acyltransferase activity from Escherichia coli producing the 40-kDa protein accepted either 6-aminopenicillanic acid or IPN as the substrate and made a penicillinase-sensitive antibiotic in the presence of phenylacetyl coenzyme A. Therefore, a single gene is responsible for converting IPN to penicillin G. The active form of the enzyme may result from processing of the 40-kDa monomeric precursor to a heterodimer containing subunits of 11 and 29 kDa.

This study was conducted to characterize and explore the endophytic fungi of Withania somnifera plant for their bioactive potential. A total of 70 segments of W. somnifera were screened and the highest species richness, as well as frequency of colonization was found in root determined as 25%. Endophytic fungi shown significant antibacterial activity against Staphylococcus aureus, E.coli, Proteus sp, Klebsiella pneumoniae and Pseudomonas sp. and were identified as Corynespora cassicola, Fusarium solani and Penicillium chrysogenum. The minimum inhibitory concentration (MIC) by the micro dilution broth was in the range of 21-82 μg/mL. Isolates revealed the presence of different phytochemicals, which includes, terpenoids, tannins, alkaloids and flavonoids. Based on its antibacterial, antioxidant and cytotoxic potential, Fusarium solani was selected, characterized genotypically by 28S rDNA typing and the sequence was deposited in NCBI database under accession number KJ193849. Fusarium solani was found to possess higher relative percentage reducing power of about 90.62% than that of Corynespora cassicola (57.47%) and Penicillium chrysogenum (52.37%) and the absorbance value obtained is comparable with the control. Percentage cell viability on cultured HeLa cell lines was determined as 6.25 μg/mL (64.74%), 12.5 μg/mL (61.08%), 25 μg/mL (58.68%), 50 μg/mL (53.24%), and 100 μg/mL (49.94%).There was significant reduction in the viability of cancer cells at 100 μg/mL. GC-MS analysis of the HPTLC fractions of Fusarium solani showed a wide range of compounds and the compounds taraxasterol, acenaphthylene, 3,4-oxadiazole, benzenamine were pharmaceutically important.

Combination of DNA barcoding, targeted metabolite profiling and multispectral imaging to identify mold species and metabolites in sliced bread

This study demonstrates the removal of fly ash with Penicillium chrysogenum, a newly isolated species of fungus, and acute toxicity assessment with Daphnia magna. In the study, two different removal mechanisms were compared, both bio-removal and bio-sorption. Six different ash and three different biomass concentrations were used simultaneously. Although other fungal species in the literature failed at such a high concentration of fly ash, P. chrysogenum was able to tolerate it even at 10% concentration. The highest bio-removal yield was recorded as 100% at 0.5% fly ash concentration. Maximum bio-sorption yield was 95.27% after 24th hour. The evaluation results of fly ash bio-toxicity by D. magna showed that the no observed effect level (NOEL) was 0.2 mg/L and the low observed effect level (LOEL) was 0.5 mg/L. The element analysis, determined by ED-XRF, clarified that Ca, Si, Fe and S were the common elements in this ash. This is the first study in the literature where fly ash removal was carried out using P. chrysogenum for both bio-removal and bio-sorption and needs to be developed in the future.