Titer improvement of mycophenolic acid in the novel producer strain Penicillium arizonense and expression analysis of its biosynthetic genes.

Mycophenolic acid (MPA) is the active ingredient in the most important immunosuppressive pharmaceuticals. It has antifungal, antibacterial, antiviral, anti-psoriasis, and antitumor activities. Therefore, its overproduction in addition to gene expression analysis was our main target. Through this study, we isolated a novel potent mycophenolic acid (MPA) producer strain of the genus Penicillium from the refrigerated Mozzarella cheese and it was identified with the molecular marker ITS and benA genes as P. arizonenseHEWt1. Three MPA overproducer mutants were isolated by exposing the wild type to different doses of gamma-rays, and the fermentation conditions for the highest production of MPA were optimized. The results indicated that MPA amounts produced by the mutants MT1, MT2, and MT3 were increased by 2.1, 1.7, and 1.6-fold, respectively, compared with the wild-type. The growth of both mutant and wild-type strains on PD broth, adjusted to pH 6 and incubated at 25 °C for 15 d, were the best conditions for maximum production of MPA. In a silico study, five orthologs genes of MPA biosynthesizing gene clusters in P. brevicompactum were predicted from the genome of P. arizonense. Sequencing and bioinformatic analyses proved the presence of five putative genes namely mpaA, mpaC, mpaF, mpaG, and mpaH in the P. arizonense HEWt1 genome. Gene expression analysis by qRT-PCR indicated an increase in the transcription value of all annotated genes in the three mutants over the wild type. A highly significant increase in the gene expression of mpaC, mpaF, and mpaH was observed in P. arizonense-MT1 compared with wild-type. These results confirmed the positive correlation of these genes in MPA biosynthesis and are the first report regarding the production of MPA by P. arizonense.Kew word.Mycophenolic acid, Penicillium arizonense, mutagenesis, gene expression.

Iron stress response and bioaccumulation potential of three fungal strains isolated from sewage-irrigated soil.

AIMS: Contamination with heavy metal (HM) is a severe environmental issue. Therefore, there is a pressing need to create environmentally safe and cost-effective HM bioremediation approaches. METHODS AND RESULTS: Three iron-tolerant fungal strains were isolated from sewage-irrigated soils, molecularly identified and deposited in the GenBank as Aspergillus flavus MT639638, A. terreus MT605370 and Fusarium oxysporum MT605399. The fungal growth, minimum inhibitory concentration (MIC), tolerance index (TI), removal efficiency, bioaccumulation, and enzymatic and non-enzymatic antioxidants were determined. Based on MIC values, A. flavus MT639638 was the most resistant strain. F. oxysporum displayed the highest percent removal efficiency (93.65% at 4000 mg L-1 ) followed by A. flavus (92.92%, at 11,000 mg L-1 ), and A. terreus (91.18% at 3000 mg L-1 ). F. oxysporum was selected based on its highly sensitivity for further characterization of its response to Fe(II) stress using TEM, SEM and EDX, in addition to HPLC analysis of organic acids. These analyses demonstrated the localization of bioaccumulated Fe(II) and ultrastructural changes induced by iron and indicated induction release of organic acids. CONCLUSIONS: Our fungal strains showed an effective capacity for removal of Fe(II) via bioaccumulation and biosorption mechanisms which were supported by instrumental analyses. The iron tolerance potentiality was mediated by induction of selected antioxidative enzymes and biomolecules. SIGNIFICANCE AND IMPACT OF THE STUDY: This study depicts a potential utilization of the three fungal strains for the bioremediation of iron-contaminated soils.

Facile synthesis of magnetic disinfectant immobilized with silver ions for water pathogenic microorganism's deactivation.

One-pot synthesis of a new magnetic disinfectant was achieved through the polymerization of thiourea and formaldehyde in the presence of magnetite nanoparticles (MTUF). The obtained magnetic chelating resin was loaded with Ag(I) ions. This material was tested as a disinfectant for water pathogenic microorganism's deactivation. The toxicity of MTUF before and after Ag(I) loading was estimated. The antimicrobial activity tests of MTUF-Ag were carried out against Escherichia coli, Salmonella Typhimurium, and Pseudomonas aeruginosa as examples of Gram-negative bacteria; Listeria monocytogenes, Staphylococcus aureus, Enterococcus faecalis, and Bacillus subtilis as examples of Gram-positive bacteria; and Candida albicans as representative for fungi. The results showed that the minimum inhibitory dosage (MID) of MTF-Ag against Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, Staphylococcus aureus, and mixed culture were 1.5, 2.0, 1.0, 1.5, and 1.5 mg/mL, respectively, after 40 min of contact time. While C. albicans was more resistant to the magnetic disinfectant, only three log reductions were done at 2.5 mg/mL. The studied MTUF-Ag was successfully tested for water and wastewater pathogenic microorganism's deactivation. It can be concluded that MTUF-Ag could be a good candidate for water disinfection.

Improved production of kojic acid by mutagenesis of Aspergillus flavus HAk1 and Aspergillus oryzae HAk2 and their potential antioxidant activity.

Two wild-type (WT) Aspergillus strains, A. flavus HAk1 and A. oryzae HAk2, were selected for kojic acid (KA) biosynthesis. Malt extract sucrose culture medium (MES) was the best culture medium for maximum production of KA. The maximum production of KA has been estimated at pH 4 after 7 days of incubation at 30 °C. Overproduction of KA was attained by mutagenesis of both A. flavus HAk1 and A. oryzae HAk2 through their exposer to different doses of gamma irradiation. The mutant strains (MT) A. flavus HAk1-M2 and A. oryzae HAk2-M26 were the most stable mutants for maximum production of KA through four generations. Yield of KA by A. oryzae HAk2-M26 and A. flavus HAk1-M2 has been 2.03-fold and 1.9-fold, respectively, higher than their wild-type strains. All WT and MT strains were used for KA production from different agricultural raw materials. Apple peel was the best waste for KA production by WT strains of A. flavus and A. oryzae, while orange peel and rice stalk are best material for KA production by MT strains, A. flavus HAk1-M2 and A. oryzae HAk2-M26, respectively. All experimental strains have the ability to produce considerable amounts of KA from sugarcane molasse (SCM) and sugar-beet molasse (SBM). SBM was better than SCM for KA production by all strains. The antioxidant activity of biosynthesizing KA was strongly affected with production conditions, where the highest antioxidant activity of all strains was recorded at the optimum environmental and nutritional conditions for KA production.