Penicillium Janthinellum Research Articles

Concurrent methylation and demethylation of arsenic by fungi and their differential expression in the protoplasm proteome

Microbial methylation and demethylation are central to arsenic's (As) biogeochemical cycling. Here, the transformations of monomethylarsonic acid (MMA(V)) (50 mg L−1) for 15 days in cells of As-methylating fungi, Fusarium oxysporum CZ-8F1, Penicillium janthinellum SM-12F4, and Trichoderma asperellum SM-12F1, were evaluated, and trace concentrations of As(III) and As(V) were observed in fungal cell extracts. Trace amounts of DMA(V) were also detected in MMA(V) and P. janthinellum SM-12F4 incubations. In situ X-ray absorption near edge structure (XANES) indicated that after exposure to MMA(V) (500 mg L−1) for 15 days, 28.6–48.6% of accumulated As in fungal cells was DMA(V), followed by 18.4–30.3% from As(V), 0–28.1% from As(III), and 4.8–28.9% from MMA(V). The concurrent methylation and demethylation of As occurs in fungal cells. Furthermore, a majority of proteins involved in metabolism, transport, ATP activity, biosynthesis, signal transduction, DNA activity, translation, and oxidative stress were upregulated in T. asperellum SM-12F1 cells after MMA(V) exposure compared to As(III), As(V), and DMA(V). The detoxification process of T. asperellum SM-12F1 was As species-specific. Methylenetetrahydrofolate reductase (R7YMH0) donation of a methyl group for S-adenosylmethionine (SAM) generation significantly increased following MMA(V) exposure.

Penicilones A-D, Anti-MRSA Azaphilones from the Marine-Derived Fungus Penicillium janthinellum HK1-6.

Four new azaphilones, penicilones A-D (1-4), were isolated from the mangrove rhizosphere soil-derived fungus Penicillium janthinellum HK1-6. Their planar structures and absolute configurations were determined by extensive analysis of NMR spectroscopic data, ECD spectra, the modified Mosher's method, and chemical conversions. Interestingly, 1 and 2 had the opposite configuration at C-7 compared to the closely related chloro analogues 3 and 4. Ester hydrolysis of 2 and 4 afforded their parental azaphilones, named penicilones E (5) and F (6). Compounds 1-6 were evaluated for their antibacterial activities in vitro. Penicilones B-D (2-4) showed potent anti-MRSA (Staphylococcus aureus ATCC 43300, ATCC 33591) activities with MIC values ranging from 3.13 to 6.25 μg/mL.

Biotransformation of capsaicin by Penicillium janthinellum AS 3.510

Biocatalysis of capsaicin (1) was performed by Penicillium janthinellum AS 3.510. Nine metabolites including four new compounds were afforded, and their structures were elucidated as (8S)-trans-8-hydroxy-8-hydroxymethyl-N-vanillyl-6-nonenamide (2), 6-hydroxy-8-methyl-N-vanillyl-7-nonenamide (3), trans-8-methoxy-8-methyl-N-vanillyl-6-nonenamide (4), 6-methoxy-8-methyl-N-vanillyl-7-nonenamide (5), dihydrocapsaicin (6), ω-1-hydroxydihydrocapsaicin (7), ω-1-hydroxycapsaicin (8), ω-hydroxycapsaicin (9), N-(4-hydroxy-3-methoxybenzyl)-5-[3-(propan-2-yl)oxiran-2-yl]pentanamide (10) by 1D and 2D NMR and HRESIMS spectra. The biotransformation processes include hydroxylation, methylation, reduction, and epoxylation.

Trichodermamides D–F, heterocyclic dipeptides with a highly functionalized 1,2-oxazadecaline core isolated from the endophytic fungus Penicillium janthinellum HDN13-309

Six trichodermamides, including three new ones, were isolated from an endophytic fungus, and two of them could function as antioxidants.

Penicisulfuranols A-F, Alkaloids from the Mangrove Endophytic Fungus Penicillium janthinellum HDN13-309.

Six new epipolythiodioxopiperazine (ETP) alkaloids, penicisulfuranols A-F (1-6), were isolated from the mangrove endophytic fungus Penicillium janthinellum HDN13-309. All structures including absolute configurations were elucidated on the basis of comprehensive spectroscopic data and ECD calculations. They belong to the unusual family of ETPs containing sulfur atoms on both α- and β-positions of amino acid residues and a rare 1,2-oxazadecaline core moiety. In addition, compounds 1-6 also possess a rare spiro-furan ring and 1-3 showed cytotoxicity with IC50 values ranging from 0.1 to 3.9 μM.

Induction of cellulases by disaccharides or their derivatives in Penicillium janthinellum EMS-UV-8 mutant

ABSTRACTDisaccharides or their derivatives were analyzed for the induction of cellulases by Penicillium janthinellum EMS-UV-8 mutant. Among seven (Crocin, Gentiobiose, Cellobiose, α-D-Cellobiose octaacetate, Trehalose, Melibiose, Turanose) different compounds tested, α-D-Cellobiose octaacetate (COA) was found to be promising for induction of cellulases. The maximum of 0.8 IU/mL filter paper units, 4.7 IU/mL β–glucosidase and 1.0 mg/mL protein were produced using 0.1% (weight/volume) COA as an inducer and 1% glucose as a carbon source for growth. Next to COA the gentiobiose, trehalose, melibiose and turanose also induce cellulase but at a low level. The induction of cellulase was also analyzed in the presence and absence of light and it was found that the absence of light supports more growth and less cellulase induction, while in the presence of light mycelial growth was restricted but more cellulases were induced. The knowledge of cellulase induction by COA and light could be beneficial for large-scale cellulase production for conversion of cellulosic biomass to biofuels.

Antimicrobial, anti-inflammatory activity and cytotoxicity of Funtumia africana leaf extracts, fractions and the isolated methyl ursolate

Funtumia africana is used to treat and manage diverse ailments including fever, inflammation, malaria, cancer and urinary incontinence in South Africa. In this study, the antibacterial, antifungal, anti-inflammatory activities and cytotoxicity of the crude extracts, fractions and an isolated compound were determined. Serial microplate dilution and bioautography methods were used to determine the antimicrobial activities. The bacteria tested were ATCC reference strains of Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa and Staphylococcus aureus. The fungal test organisms used were three clinical isolates (Aspergillus fumigatus, Cryptococcus neoformans and Candida albicans), C. albicans ATCC 10231 and three phytopathogenic fungi (Fusarium oxysporum, Penicillium janthinellum and Rhizoctonia solani). The anti-inflammatory activity was determined using cyclooxygenase (COX) enzymes and the MTT assay was used to determine cellular toxicity against Vero and human liver (C3A) cells. The crude extract had MICs as low as 80μg/ml against both bacteria and fungi. The chloroform fraction had the lowest MIC of 20μg/ml against P. aeruginosa. The hexane and chloroform fractions had MIC of 40μg/ml against C. albicans ATCC 10231. The crude extract, hexane and chloroform fractions had moderate activity against both COX-1 and COX-2. The chloroform fraction was more active than the crude extract (59.7%) with an inhibition of 68.2% against COX-1. Bioassay-guided fractionation using column chromatography led to the isolation of methyl ursolate (MU) with an MIC of 62.5μg/ml against F. oxysporum. It was relatively toxic against Vero cells with an IC50 of 10.4μg/ml. The antimicrobial and anti-inflammatory activities of the crude extract provide some support for the traditional use of the plant.

ANTIOXIDANT AND ANTIFUNGAL ACTIVITY OF SELECTED MEDICINAL PLANT EXTRACTS AGAINST PHYTOPATHOGENIC FUNGI.

Background:Medicinal plants are used by many ethnic groups as a source of medicine for the treatment of various ailments in both humans and domestic animals. These plants produce secondary metabolites that have antimicrobial properties, thus screening of medicinal plants provide another alternative for producing chemical fungicides that are relatively non-toxic and cost-effective.Materials and methods:Leaf extracts of selected South African plant species (Bucida buceras, Breonadia salicina, Harpephyllum caffrum, Olinia ventosa, Vangueria infausta and Xylotheca kraussiana) were investigated for activity against selected phytopathogenic fungi (Aspergillus niger, Aspergillus parasiticus, Colletotricum gloeosporioides, Penicillium janthinellum, P. expansum, Trichoderma harzianum and Fusarium oxysporum). These plant fungal pathogens causes major economic losses in fruit industry such as blue rot on nectaries and postharvest disease in citrus. Plant species were selected from 600 evaluated inter alia, against two animal fungal pathogens (Candida albicans and Cryptococcus neoformans). Antioxidant activity of the selected plant extracts were investigated using a qualitative assay (2, 2-diphenyl-1-picrylhydrazyl (DPPH)). Bioautography assay was used to determine the number of antifungal compounds in plant extracts.Results:All plant extracts were active against the selected plant phytopathogenic fungi. Moreover, Bucida buceras had the best antifungal activity against four of the fungi, with minimum inhibitory concentration (MIC) values as low as 0.02 mg/ml and 0.08 mg/ml against P. expansum, P. janthinellum, T. harzianum and F. oxysporum. The plant extracts of five plant species did not possess strong antioxidant activity. However, methanol extract of X. kraussiana was the most active radical scavenger in the DPPH assay amongst the six medicinal plants screened. No antifungal compounds were observed in some of the plant extracts with good antifungal activity as shown in the microdilution assay, indicating possible synergism between the separated metabolites.Conclusion:The results showed that acetone was the best extractant. Furthermore, our findings also confirm the traditional use of Breonadia salicina and demonstrate the potential value of developing biopesticides from plants.

Production, purification and characterization of an exo-polygalacturonase from Penicillium janthinellum sw09

A soil isolate, Penicillium janthinellum sw09 has been found to produce significant amounts of an extracellular pectinase subsequently characterized as exo-polygalacturonase (exo-PG). By optimizing growth conditions, P. janthinellum sw09 produced high amount of exo-PG (16.54 units/mL). The crude enzyme was purified by gel filtration chromatography and two exo-PG activity peaks (designated as PGI and PGII) were revealed. On SDS-PAGE analysis, purified PGII using DEAE-Sepharose FF column, was found to be a single band with a molecular mass of 66.2 kDa. The purified PGII exhibited maximal activity at the temperature of 45 oC and pH 5.0. The stability profiles show that PGII is more stable in the pH range of 4.0-8.0 and below 60 oC. The Km and Vmax for the enzyme was 1.74 mg/mL and 18.08 μmol/ (mL•min), respectively. Due to this enzymatic characterization, this pectinase is an attractive candidate for applications in degradation of pectin.

Chemical characterization of fatty acids, alkanes, n-diols and alkyl esters produced by a mixed culture of Trichoderma koningii and Penicillium janthinellum grown aerobically on undecanoic acid, potatoe dextrose and their mixture

ABSTRACTLittle is known about the mixed fungal synthesis of high-value aliphatics derived from the metabolism of simple and complex carbon substrates. Trichoderma koningii and Penicillium janthinellum were fed with undecanoic acid (UDA), potatoe dextrose broth (PDB), and their mixture. Pyrolysis Field Ionization Mass Spectrometry (Py-FIMS) together with 1H and 13C Nuclear Magnetic Resonance (NMR) characterized CHCl3 soluble aliphatics in the fungal cell culture. Data from NMR and Py-FIMS analysis were complementary to each other. On average, the mixed fungal species produced mostly fatty acids (28% of total ion intensity, TII) > alkanes (2% of TII) > n-diols (2% of TII) > and alkyl esters (0.8% of TII) when fed with UDA, PDB or UDA+PDB. The cell culture accumulated aliphatics extracellularly, although most of the identified compounds accumulated intracellularly. The mixed fungal culture produced high-value chemicals from the metabolic conversion of simple and complex carbon substrates.

Improvement of wheat straw hydrolysis by cellulolytic blends of two Penicillium spp.

Co-culture of fungal strains Penicillium janthinellum EMS-UV-8 (E), Penicillium funiculosum strain P (P) and Aspergillus sp. strain G (G) and blending of their crude cellulase were evaluated for improvements in cellulase activities as well as for enhanced hydrolysis of dilute acid pretreated wheat straw (PWS). The blending of crude enzymes of P and E enhanced the hydrolysis of PWS more effectively due to synergism in cellulolytic enzyme activities. Here, three types of blends were made on the basis of equal FPUs, equal protein content or fixed volume containing different proportions of individual enzymes, the former blend hydrolyzed 42.6% of PWS due to the 98%,62%, 64% and 34% synergistic enhancement in endo-glucanase, cellulase (FPU), β-glucosidase and xylanase activities, respectively. Hydrolysis at 10% solid loading of PWS in roller bottle reactor with this blend further enhanced hydrolysis yield to 74% within 24 h, which was much better than the corresponding hydrolysis yields of individual (38.1% by E and 61.5% by P) or the commercial enzyme (62.3%). This study proved that synergistic blends of cellulases from two Penicillium spp. are cost-effective tools for efficient wheat straw hydrolysis for on-site biofuel production.

Self-immobilization of a magnetic biosorbent and magnetic induction heated dye adsorption processes

Biosorbents for wastewater treatments is one of the most widely studied techniques in environmental and bioresource research. A marine-derived Penicillium janthinellum strain fungus was used in this study to prepare self-immobilized mycelial pellets. Fe3O4 nanoparticles were added in the culturing media, which were automatically incorporated in the mycelial pellets during culturing process to endow the pellets with magnetic properties. These magnetic mycelial pellets were used as biosorbents for water treatments, and their morphology was characterized by microscopic observation, Energy dispersive X-ray analysis, Fourier transform infrared spectroscopy and thermal gravimetric analysis. The effects of nanoparticles in culture media on pellet formation were discussed and the content of nanoparticle in culturing media was optimized. Study on Congo red adsorption from aqueous solutions showed that the pellets with 71.6wt% Fe3O4 nanoparticles had a maximum adsorption capacity of 102.4mg/g pellets (360.6mg/g mycelia), and the solution heated by magnetic induction showed faster adsorption rates than that of bulk liquid heating processes under the same adsorption temperature. These pellets were easily separated by magnetic forces and the magnetic induction heating is beneficial in energy saving when comparing to traditional heating processes.

Separation of a compound effective against Biomphalaria alexandrina snails from the filtrate of Penicillium janthinellum

Biomphalaria alexandrina snails, as intermediate hosts of schistosomiasis, play a central role in dissemination of the disease. Control of these snails by chemical molluscicides adversely affects the aquatic environment, causing toxic and carcinogenic effects on non-target organisms. Searching for promising substances from biological origin becomes an urgent need to overcome these drawbacks. Screening tests were carried out on 236 fungal genera isolated from the habitat of freshwater snails in four Egyptian governorates. Twenty species were effective against B. alexandrina snails, but the most potent was Penicillium janthinellum as the value of LC50 was 1.03%. Chemical analyses of this filtrate resulted in the separation of a compound effective against snails; it was identified as methyl gallate. Protein electrophoresis showed that both fungal filtrate and methyl gallate affect the protein pattern of snails’ haemolymph. Little or no mortality of Daphnia pulex individuals was observed after their exposure to sub lethal concentrations of each treatment.

Untreated wheat straw: Potential source for diverse cellulolytic enzyme secretion by Penicillium janthinellum EMS-UV-8 mutant

Study describes the production of cellulases by Penicillium janthinellum EMS-UV-8 using untreated wheat straw (WS), treated WS (acid, alkali, steam exploded, organo-solv) and pure cellulosic substrates (avicel, cellulose-II and carboxymethyl cellulose). Severely pretreated WS and cellulose-II produced more cellulolytic enzymes than untreated samples. XRD and FTIR analysis revels that the increase in the amorphous structure of pretreated WS/cellulose increases enzyme production. Enzyme samples prepared using different substrates were used for the hydrolysis of dilute acid treated wheat straw (DATWS), steam exploded wheat straw (SEWS) and avicel. The enzyme prepared using untreated WS gave more hydrolysis of DATWS and SEWS than the enzyme prepared using pretreated WS or pure cellulosic substrates. This revels that more diverse/potential enzymes were secreted by P. janthinellum EMS-UV-8 mutant using untreated WS. This study may contribute in production of efficient enzyme mixture/cocktail by single fungal strain for economic conversion of biomass to sugars.

Paths and determinants for Penicillium janthinellum to resist low and high copper.

Copper (Cu) tolerance was well understood in fungi yeasts but not in filamentous fungi. Filamentous fungi are eukaryotes but unlike eukaryotic fungi yeasts, which are a collection of various fungi that are maybe classified into different taxa but all characterized by growth as filamentous hyphae cells and with a complex morphology. The current knowledge of Cu resistance of filamentous fungi is still fragmental and therefore needs to be bridged. In this study, we characterized Cu resistance of Penicillium janthinellum strain GXCR and its Cu-resistance-decreasing mutants (EC-6 and UC-8), and conducted sequencing of a total of 6 transcriptomes from wild-type GXCR and mutant EC-6 grown under control and external Cu. Taken all the results together, Cu effects on the basal metabolism were directed to solute transport by two superfamilies of solute carrier and major facilitator, the buffering free CoA and Acyl-CoA pool in the peroxisome, F-type H+-transporting ATPases-based ATP production, V-type H+-transporting ATPases-based transmembrane transport, protein degradation, and alternative splicing of pre-mRNAs. Roles of enzymatic and non-enzymatic antioxidants in resistance to low and high Cu were defined. The backbone paths, signaling systems, and determinants that involve resistance of filamentous fungi to high Cu were determined, discussed and outlined in a model.

Crop Residue Degradation by Autochthonous Fungi Isolated from Cropping System Management Scenarios

In the rice-wheat system dominated belt of India (Indo-Gangetic plains; IGP), paddy leaves, about 8 to 9 t/ha of loose and anchored residue in the field, are mechanically harvested. Farmers prefer to burn this residue to clear the field for the timely preparation of conventional wheat sowing. In-situ degradation by autochthonous fungi can be a better option for the disposal of unwanted crop residues. Autochthonous fungi isolated from conservation agriculture-based crop management fields were screened and assessed for their residue degradation potential. Nineteen isolates were selected for detailed enzymatic analysis in submerged fermentation, responsible for lignocellulosic residue degradation. Out of these five fungal isolates RPW 1/3 (Aspergillus flavus), RPW 1/6 (Aspergillus terreus), RPW 1/9 (Aspergillus terreus), RPWM 2/2 (Penicillium janthinellum) and RZWM 3/1 (Aspergillus niger) showed higher activities of cellobiase, CMCase, FPase, xylanase, and laccase enzymes in solid state fermentation. Further two isolates RPW 1/3 and 1/6 showed approximately 30% degradation of straw residue after 10 days incubation.

Biotransformation of alisol G by Penicillium janthinellum and the hCE2 inhibitory effects of its metabolites

Alisol G, also named 25-anhydroalisol A, is a major triterpene obtained from dried rhizomes of Alisma orientalis. This paper investigated the transformation of alisol G by fungi and evaluated the hCE2 inhibitory effects of the transformed products. By screening whole cells of 10 strains of filamentous fungi, Penicillium janthinellum AS 3.510 exhibited a special capability to transform alisol G. Four metabolites were obtained, which were determined to be new compounds on the basis of spectroscopic data, including UV, 1D- and 2D-NMR, and HRESIMS. In addition, in an in vitro bioassay, metabolites 1–4 showed significant hCE2 inhibitory activities with IC50 values of 6.81, 16.66, 3.38, and 6.33 μM, respectively.

Demethylation of arsenic limits its volatilization in fungi

Arsenic (As) biomethylation is increasingly being regarded as a promising method to volatize As from the environment; however, the As volatilization efficiency of most microorganisms is low. Here, the speciation transformation of dimethylarsinic acid (DMA) as an important methylation intermediate in the cells of Fusarium oxysporum CZ-8F1, Penicillium janthinellum SM-12F4, and Trichoderma asperellum SM-12F1 were investigated. These fungal strains have been certified to volatilize As from As-loaded environment. In situ X-ray absorption near edge structure (XANES) indicated that demethylation of DMA with methylarsonic acid (MMA), arsenate [As(V)], and arsenite [As(III)] as intermediates or products occurred in fungal cells after exposure to DMA for 15 days. 36.7–55.7% of the original DMA could lose one or two methyl groups and be changed into MMA or inorganic As. Chromatographic separation of the cell lysates also supported these findings. Thus it comes that demethylation might be a remarkable internal factor limiting As volatilization efficiency.

Phytohormones enabled endophytic fungal symbiosis improve aluminum phytoextraction in tolerant Solanum lycopersicum: An examples of Penicillium janthinellum LK5 and comparison with exogenous GA3

This work investigates the potentials of fungal-endophyte Penicillium janthinellum LK5 (PjLK5) and its inherent gibberellic acid (GA3) as reference to enhance aluminum (Al) induced toxicity in tolerant tomato (Solanum lycopersicum) plants. Initial screening showed significantly higher uptake of Al by PjLK5. Aluminum stress (100μM) significantly retarted plant growth in control plants. Conversely PjLK5 and GA3 application significantly increased morphological attributes of Al-tolerant tomato plants with or without Al-stress. PjLK5 inoculation with and without Al-stress maintained the plant growth whilst extracting and translocating higher Al in shoot (∼192mg/kg) and root (∼296mg/kg). This was almost similar in GA3 treatments as well. In addition, PjLK5 inoculation extended protective effects to tomato plants by maintaining reduced cellular superoxide anions in Al stress. Al-induced oxidative stress was further reduced due to significantly higher activity of metal-responsive reduced glutathione. The functional membrane was less damaged in PjLK5 and GA3 treatments because the plants synthesized reduced levels of malondialdhyde, lenolenic and linoleic acids. Defense-related endogenous phytohormone salicylic acid was significantly up-regulated to counteract the adverse effects of Al-stress. In conclusion, the PjLK5 possess a similar bio-prospective potential as of GA3. Application of such biochemically active endophyte could increase metal phytoextraction whilst maintaining crop physiological homeostasis.

Selection and molecular characterization of cellulolytic–xylanolytic fungi from surface soil-biomass mixtures from Black Belt sites

Plant biomass is an abundant renewable natural resource that can be transformed into chemical feedstocks. Enzymes used in saccharification of lignocellulosic biomass are a major part of biofuel production costs. A cocktail of cellulolytic and xylanolytic enzymes are required for optimal saccharification of biomass. Accordingly, thirty-two fungal pure cultures were obtained from surface soil-biomass mixtures collected from Black Belt sites in Alabama by culturing on lignocellulosic biomass medium. The fungal strains were screened for the coproduction of cellulolytic and xylanolytic enzymes. Strains that displayed promising levels of cellulolytic and xylanolytic enzymes were characterized by molecular analysis of DNA sequences from the large subunit and internal transcribed spacer (ITS) of their ribosomal RNA gene. Nucleotide sequence analysis revealed that two most promising isolates FS22A and FS5A were most similar to Penicillium janthinellum and Trichoderma virens. Production dynamics of cellulolytic and xylanolytic enzymes from these two strains were explored in submerged fermentation. Volumetric productivity after 120h incubation was 121.08units/L/h and 348units/L/h for the filter paper cellulase and xylanase of strain FS22A, and 90.83units/L/h and 359units/L/h, respectively for strain FS5A. Assays with 10 times dilution of enzymes revealed that the activities were much higher than that observed in the crude culture supernatant. Additionally, both FS22A and FS5A also produced amylase in lignocellulose medium. The enzyme profiles of these strains and their activities suggest potential applications in cost effective biomass conversion and biodegradation.