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Abstract Background Low-cost cellulase production is vital to sustainable second-generation biorefineries. The catabolically derepressed strain of Penicillium funiculosum NCIM1228 (PfMig188 or ∆Mig1) secretes a superior set of cellulolytic enzymes, that are most suitable for 2G biorefineries. At a 3% (w/w) load, the ∆Mig1 secretome can release > 80% of fermentable sugars from lignocellulose at a 15% (w/v) biomass load, irrespective of the type of biomass and pretreatment. The robustness of the secretome can be further increased by improving the cellulase production capacity of the fungal strain. Results We began by identifying the transcription factor responsible for cellulase production in NCIM1228. An advanced RNA-seq screen identified three genes, clr-2, ctf1a and ctf1b; the genes were cloned under their native promoters and transformed into NCIM1228. Of the three, clr-2 overexpression led to twofold higher cellulase production than the parent strain and was thus identified as the transcriptional activator of cellulase in NCIM1228. Next, we overexpressed clr-2 in ∆Mig1 and expected an exponential increase in cellulolytic attributes accredited to the reinforced activation mechanisms, conjoint with diminished negative regulation. Although clr-2 overexpression increased the transcript levels of cellulase genes in ∆Mig1, there was no increase in cellulase yield. Even a further increase in the transcript levels of clr-2 via a stronger promoter was ineffective. However, when the CaCO3 concentration was increased to 5 g/l in the growth medium, we achieved a 1.5-fold higher activity of 6.4 FPU/ml in the ∆Mig1 strain with clr-2 overexpression. Enthused by the calcium effect, a transcriptomic screen for genes encoding Ca2+-activated kinase identified ssp1, whose overexpression could further increase cellulase yield to ~ 7.5 FPU/ml. Investigation of the mechanism revealed that calcium signaling exclusively enhances the translation and secretion of cellulase in Penicillium funiculosum. Conclusions Our study identifies for the first time that cellulose activates two discrete signaling events to govern cellulase transcription and posttranscriptional processes (translation, processing and secretion) in P. funiculosum NCIM1228. Whereas Clr-2, the transcriptional activator of cellulase, governs transcription, calcium signaling specifically activates cellulase translation and secretion.

Background: Low-cost cellulase production is vital to sustainable second-generation biorefineries. The catabolically derepressed strain of Penicillium funiculosum NCIM1228 (PfMig188 or ∆Mig1) secretes a superior set of cellulolytic enzymes, that are most suitable for 2G biorefineries. At a 3% (w/w) load, the ∆Mig1 secretome can release > 80% of fermentable sugars from lignocellulose at a 15% (w/v) biomass load, irrespective of the type of biomass and pretreatment. The robustness of the secretome can be further increased by improving the cellulase production capacity of the fungal strain. Results: We began by identifying the transcription factor responsible for cellulase production in NCIM1228. An advanced RNA-seq screen identified three genes, clr-2, ctf1a and ctf1b; the genes were cloned under their native promoters and transformed into NCIM1228. Of the three, clr-2 overexpression led to twofold higher cellulase production than the parent strain and was thus identified as the transcriptional activator of cellulase in NCIM1228. Next, we overexpressed clr-2 in ∆Mig1 and expected an exponential increase in cellulolytic attributes accredited to the reinforced activation mechanisms, conjoint with diminished negative regulation. Although clr-2 overexpression increased the transcript levels of cellulase genes in ∆Mig1, there was no increase in cellulase yield. Even a further increase in the transcript levels of clr-2 via a stronger promoter was ineffective. However, when the CaCO3 concentration was increased to 5 g/l in the growth medium, we achieved a 1.5-fold higher activity of 6.4 FPU/ml in the ∆Mig1 strain with clr-2 overexpression. Enthused by the calcium effect, a transcriptomic screen for genes encoding Ca2+-activated kinase identified ssp1, whose overexpression could further increase cellulase yield to ~ 7.5 FPU/ml. Investigation of the mechanism revealed that calcium signaling exclusively enhances the translation and secretion of cellulase in Penicillium funiculosum. Conclusions: Our study identifies for the first time that cellulose activates two discrete signaling events to govern cellulase transcription and posttranscriptional processes (translation, processing and secretion) in P. funiculosum NCIM1228. Whereas Clr-2, the transcriptional activator of cellulase, governs transcription, calcium signaling specifically activates cellulase translation and secretion. [ABSTRACT FROM AUTHOR]

The cellulase enzyme is currently the world's third largest commercial enzyme. Because of its requirement in various industries like textiles, food, waste management, pharmaceuticals, agriculture, pulp paper, biofuels, and others, its demand curve has increased sharply. The response surface methodology (RSM) approach was employed to optimize media components and process parameters in the current investigation, which successfully increased the cellulase production from Penicillium funiculosum NCIM 1228. Statistical optimization for the hyperproduction of cellulases was conducted using RSM. The Plackett–Burman design (PBD) approach was used to investigate the critical factors of the cellulase production medium. Subsequently, the Box–Behnken design (BBD) method was used to statistically estimate optimum values and conditions that substantially impacted cellulase production. The estimated optimal combinations of parameters for cellulase production were urea (0.2%), CaCl2 (0.2%), MgSO4 (0.05%), peptone (1.5%), microcrystalline cellulose (5.0%), wheat bran (2.5%), corn steep liquor (CSL) (2.5%), KH2PO4 (0.15%), inoculum (10.65%), agitation (157 rpm), pH (5.88), and temperature (29.84 °C). Conclusively, experimental validation under optimal conditions detected an increased production of 3.82- and 3.61-fold in filter paper assay (FPase) and β-glucosidase, respectively. Additionally, 1.66- and 1.57-fold enhancement in FPase and β-glucosidase specific activity was observed where an xylanase activity was enhanced by 3.29-fold. Furthermore, the enzyme showed 51.30 per cent hydrolysis efficiency on sugarcane bagasse lignocellulosic biomass (LCB), at a dose of 7 FPase units per g of cellulose. P. funiculosum NCIM 1228 offers the benefit of producing cellulase with an entire cellulolytic system of enzymes that can be synthesized extracellularly, thus acting as a promising biocatalyst for biofuel industries. [ABSTRACT FROM AUTHOR]

Introduction[1]: Sunflower meal is considered as an attractive alternative to soybean meal due to its adaptation to dry and temperate climates as well as its lower anti-nutrient content than other oilseeds. Sunflower meal has a limiter that restricts its use in broiler diets. The fiber in sunflower meal has limited its use in the diet of broilers due to reduced metabolic energy as well as production. Fermentation, which uses beneficial microorganisms, can reduce the chemical composition of the feed and increase the yield of broilers by producing enzymes and using fiber as a feed source. In this study, in order to investigate the effect of fermentation on sunflower meal, growth yield, microbial population composition, blood parameters in broiler chickens were performed. Materials and methods: The research was conducted at the research farm of Sari University of Agricultural Sciences and Natural Resources in 2020. In this study, 200 commercial male Ross 308 broilers were used in a completely randomized design with five treatments, five replications, and eight chickens per replication. The treatments included: 1) Control diet, 2) Diet containing sunflower meal (negative control), 3) Diet containing sunflower meal fermented with Aspergillus Oryzae, 4) Diet containing sunflower meal fermented with Penicillium Funiculosum, and 5) Diet containing sunflower meal fermented with Aspergillus Oryzae and Penicillium Funiculosum. Sunflower meal was obtained from Behpak Behshahr (Mazandaran) company, ground in the animal nutrition laboratory of Sari University of Agricultural Sciences and Natural Resources, and sieved to a size of 2 mm. Aspergillus Oryzae (PTCC5010) and Penicillium Funiculosum (PTCC5301) were purchased as lyophilized vials from the Fungus and Bacteria Collection Center of the Iranian Research Organization for Science and Technology (IROST). During the fermentation process, 1.1 liters of the mixture of distilled water and primer culture (containing at least 105 colony forming units per ml) was added to each kilogram of sunflower meal, and the resulting mixture was thoroughly mixed by hand for 15 minutes. The mixture was fermented in special tanks (with a one-way valve to remove the produced gases and prevent air from entering) for 7 days at 30 °C, and finally, the fermented sunflower meal was dried for three days at room temperature. All chickens were kept under the same breeding conditions during the 39-day period and had free access to feed. Experimental diets were adjusted to three periods: initial (1-10 days), growth (11-24), and final (25-39) using Table 1 of the Ross 308 Catalog of Nutritional Requirements for Broilers. The pH of fermented sunflower meal was determined using the Chiang et al. method. Sunflower meal was sampled before and after fermentation to measure dry matter, ash, crude protein by the Kjeldahl method, crude fat by Soxhlet device, and insoluble fibers in acidic detergent and neutral detergent using a fibrotec device. Total amino acid was measured by the ninhydrin method. Soluble sugar was measured by the intron method. Total phenol content was measured using the Folin-Siocalcu reagent and spectrophotometry. Soluble proteins were measured by the Bradford method. Results and discussion: During the fermentation of sunflower meal, pH, crude fat, soluble protein and soluble sugar decreased. The amount of crude protein, total amino acid, insoluble fiber in acidic detergent, and insoluble fiber in neutral detergent increased. In the whole breeding period, body weight gain was improved in treatments fed with fermented feed. In the treatment of Aspergillus­ Oryza + Penicillium funiculosum, feed consumption decreased compared to control, negative control and Penicillium funiculosum treatments. The feed conversion ratio increased in the negative control treatment at the age of 1-39 days compared to other treatments. In the ileum, coliform bacteria decreased in negative control treatments, Aspergillus Oryzae, Aspergillus Oryzae + Penicillium funiculosum compared to the control treatment, but Penicillium funiculosum treatment increased compared to other treatments. At 39 days of age, the use of fermented and raw feed increased the level of HDL-c in the negative control and Aspergillus Oryzae + Penicillium funiculosum treatments. The amount of LDL-c increased in the treatments of Aspergillus Oryzae and Penicillium funiculosum Conclusion: Based on the results, the use of fermented feed in the feeding of broiler chickens during the entire breeding period has improved weight gain and feed conversion ratio.

The enzymatic conversion of lignocellulosic biomass to bioethanol depends on efficient enzyme systems with β-glucosidase as one of the key components. In this study, we performed in-depth profiling of the various β-glucosidases present in the genome of the hypercellulolytic fungus Penicillium funiculosum using genomics, transcriptomics, proteomics, and molecular dynamics simulation approaches. Of the eight β-glucosidase genes identified in the P. funiculosum genome, three were predicted to be extracellular based on signal peptide prediction and abundance in the secretome. Among the three secreted β-glucosidases, two belonged to the GH3 family and one belonged to the GH1 family. Homology models of these proteins predicted a deep and narrow active site for the GH3 β-glucosidases (PfBgl3A and PfBgl3B) and a shallow open active site for the GH1 β-glucosidase (PfBgl1A). The enzymatic assays indicated that P. funiculosum-secreted proteins showed high β-glucosidase activities with prominent bands on the 4-methylumbelliferyl β-D-glucopyranoside zymogram. To understand the contributory effects of each of the three secreted β-glucosidases (PfBgls), the corresponding gene was deleted separately, and the effect of the deletion on the β-glucosidase activity of the secretome was examined. Although not the most abundant, PfBgl3A was found to be one of the most important β-glucosidases, as evidenced by a 42% reduction in β-glucosidase activity in the ΔPfBgl3A strain. Our results advance the understanding of the genetic and biochemical nature of all β-glucosidases produced by P. funiculosum and pave the way to design a superior biocatalyst for the hydrolysis of lignocellulosic biomass. Commercially available cellulases are primarily produced from Trichoderma reesei. However, external supplementation of the cellulase cocktail from this host with exogenous β-glucosidase is often required to achieve the desired optimal saccharification of cellulosic feedstocks. This challenge has led to the exploration of other cellulase-producing strains. The nonmodel hypercellulolytic fungus Penicillium funiculosum has been studied in recent times and identified as a promising source of industrial cellulases mainly due to its ability to produce a balanced concoction of cellulolytic enzymes, including β-glucosidases. Various genetic interventions targeted at strain improvement for cellulase production have been performed; however, the β-glucosidases of this strain have remained largely understudied. This study, therefore, reports profiling of all eight β-glucosidases of P. funiculosum via molecular and computational approaches. The results of this study provide useful insights that will establish the background for future engineering strategies to transform this fungus into an industrial workhorse. [ABSTRACT FROM AUTHOR]

Fungi cause diverse, serious socio-economic problems, including biodeterioration of valuable products and materials that spawns a biocides industry worth ~$11 billion globally. To help combat environmental fungi that commonly colonise material products, this study tested the hypothesis that combination of an approved fungicide with diverse agents approved by the FDA (Food and Drug Administration) could reveal potent combinatorial activities with promise for fungicidal applications. The strategy to use approved compounds lowers potential development risks for any effective combinations. A high-throughput assay of 1280 FDA-approved compounds was conducted to find those that potentiate the effect of iodopropynyl-butyl-carbamate (IPBC) on the growth of Trichoderma virens; IPBC is one of the two most widely used Biocidal Products Regulations–approved fungicides. From this library, 34 compounds in combination with IPBC strongly inhibited fungal growth. Low-cost compounds that gave the most effective growth inhibition were tested against other environmental fungi that are standard biomarkers for resistance of synthetic materials to fungal colonisation. Trifluoperazine (TFZ) in combination with IPBC enhanced growth inhibition of three of the five test fungi. The antifungal hexetidine (HEX) potentiated IPBC action against two of the test organisms. Testable hypotheses on the mechanisms of these combinatorial actions are discussed. Neither IPBC + TFZ nor IPBC + HEX exhibited a combinatorial effect against mammalian cells. These combinations retained strong fungal growth inhibition properties after incorporation to a polymer matrix (alginate) with potential for fungicide delivery. The study reveals the potential of such approved compounds for novel combinatorial applications in the control of fungal environmental opportunists. Key points: • Search with an approved fungicide to find new fungicidal synergies in drug libraries. • New combinations inhibit growth of key environmental fungi on different matrices. • The approach enables a more rapid response to demand for new biocides. [ABSTRACT FROM AUTHOR]

Abstract The food enzyme cellulase (4‐(1,3;1,4)‐β‐d‐glucan 4‐glucanohydrolase; EC 3.2.1.4) is produced with the non‐genetically modified Penicillium funiculosum strain Lzc35 by Danisco US Inc. The cellulase is intended to be used in distilled alcohol production, baking and brewing processes. Since residual amounts of total organic solids (TOS) are removed by distillation, dietary exposure was only calculated for baking and brewing processes. Based on the proposed maximum use levels, dietary exposure to the food enzyme–TOS was estimated to be up to 0.416 mg TOS/kg body weight (bw) per day. Genotoxicity tests did not raise a safety concern. The systemic toxicity was assessed by means of a repeated dose 90‐day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 84 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure, resulted in a margin of exposure of at least 200. Similarity of the amino acid sequence of the food enzyme to those of known allergens was searched and no match was found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions by dietary exposure cannot be excluded, but the likelihood is considered to be low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

The food enzyme cellulase (4-(1,3;1,4)-b-D-glucan 4-glucanohydrolase; EC 3.2.1.4) is produced with the non-genetically modified strain Lzc35 by Danisco US Inc. The cellulase is intended to be used in distilled alcohol production, baking and brewing processes. Since residual amounts of total organic solids (TOS) are removed by distillation, dietary exposure was only calculated for baking and brewing processes. Based on the proposed maximum use levels, dietary exposure to the food enzyme--TOS was estimated to be up to 0.416 mg TOS/kg body weight (bw) per day. Genotoxicity tests did not raise a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 84 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure, resulted in a margin of exposure of at least 200. Similarity of the amino acid sequence of the food enzyme to those of known allergens was searched and no match was found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions by dietary exposure cannot be excluded, but the likelihood is considered to be low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use. [ABSTRACT FROM AUTHOR]

Lytic polysaccharide monooxygenases (LPMOs) are crucial industrial enzymes required in the biorefinery industry as well as in the natural carbon cycle. These enzymes, known to catalyze the oxidative cleavage of glycosidic bonds, are produced by numerous bacterial and fungal species to assist in the degradation of cellulosic biomass. In this study, we annotated and performed structural analysis of an uncharacterized LPMO from Penicillium funiculosum (PfLPMO9) based on computational methods in an attempt to understand the behavior of this enzyme in biomass degradation. PfLPMO9 exhibited 75% and 36% sequence identities with LPMOs from Thermoascus aurantiacus (TaLPMO9A) and Lentinus similis (LsLPMO9A), respectively. Furthermore, multiple fungal genetic manipulation tools were employed to simultaneously overexpress LPMO and cellobiohydrolase I (CBH1) in a catabolitederepressed strain of Penicillium funiculosum, PfMig188 (an engineered variant of P. funiculosum), to improve its saccharification performance toward acid-pretreated wheat straw (PWS) at 20% substrate loading. The resulting transformants showed improved LPMO and CBH1 expression at both the transcriptional and translational levels, with -200% and -66% increases in ascorbate-induced LPMO and Avicelase activities, respectively. While the secretome of PfMig88 overexpressing LPMO or CBH1 increased the saccharification of PWS by 6% or 13%, respectively, over the secretome of PfMig188 at the same protein concentration, the simultaneous overexpression of these two genes led to a 20% increase in saccharification efficiency over that observed with PfMig188, which accounted for 82% saccharification of PWS under 20% substrate loading. [ABSTRACT FROM AUTHOR]

• The secretome of Penicillium funiculosum demonstrated potent saccharification. • Saccharification by derepressed P. funiculosum (Pf Mig188) was 2-fold higher. • Optimal saccharification was achieved at pH 4.0 and a temperature range of 45-55°C. • The Pf Mig188 secretome hydrolysed variety of alkali and acid pretreated biomasses. • The secretome demonstrated minimal product inhibition at a 20 % biomass load. PfMig188, a catabolically derepressed engineered strain of the hyper-cellulolytic fungus Penicillium funiculosum NCIM1228, was investigated for the efficacy of its secretome for biomass saccharification. An inexpensive version of media containing microcrystalline cellulose, wheat bran and soya protein was optimized for producing a high-quality secretome from the PfMig188 strain in both shake flasks and in a 20-L bioreactor. The activities of four classes of core cellulolytic enzymes required for saccharification in the PfMig188 secretome, namely, cellobiohydrolase (Avicelase activity), endoglucanase (CMCase activity), β-glucosidase (PNPGase activity) and xylanase (xylanase activity), were found to be 2.29 U/mL, 28.24 U/mL, 150 U/mL and 76 U/mL, respectively. The saccharification potential of the PfMig188 secretome was evaluated on rice straw and sugarcane bagasse pretreated with nitric acid and/or ammonium hydroxide. Saccharification performed using a 15 % (w/v) biomass load and a 3% (w/w) enzyme load released >100 g/L sugar in the hydrolysate, irrespective of the type of biomass and pre-treatment, with >80 % hydrolysis. Furthermore, the presence of lignin in nitric acid-pretreated biomass only marginally affected saccharification. Overall, the results demonstrated that the PfMig188 secretome, having relatively broad substrate specificity, is a viable and efficient substitute for T. reesei -based secretomes for diverse biomass saccharification. [ABSTRACT FROM AUTHOR]

Lân là một trong những nguyên tố chính quyết định năng suất cây trồng và nấm hòa tan lân đóng vai trò quan trọng trong gia tăng lượng lân hữu dụng cho cây trồng. Mục tiêu của nghiên cứu nhằm phân lập và định danh một số dòng nấm hòa tan lân cao từ nền đất lúa tưới nước ngập khô xen kẽ kết hợp bón phân hữu cơ ở nhà lưới. Môi trường NBRIP dùng để phân lập và đánh giá khả năng hòa tan lân. Lân hòa tan được xác định bằng phương pháp hiện màu molybdate. Kết quả cho thấy tổng cộng có 37 dòng nấm có khả năng hòa tan lân được phân lập từ 7 mẫu đất. Trong đó, hai dòng nấm ký hiệu B1 và B10 hoà tan lân lần lượt đạt 2104 mg.L-1 và 2618 mg.L-1 sau 3 và 4 ngày thí nghiệm. Hai dòng nấm này đều hòa tan lân tốt ở mức pH từ 5-7, nhiệt độ 25oC-35oC, độ mặn lên đến 0,5-1% NaCl và hòa tan tốt các dạng lân khó tan như AlPO4 và FePO4. Kết quả định danh thông qua đoạn ITS cho thấy hai dòng nấm này được định danh khoa học lần lượt là Penicillium funiculosum B1 và Aspergillus tubingensis B10. Tóm lại, hai dòng nấm này có vai trò và tiềm năng quan trọng trong việc sản xuất chế phẩm vi sinh giúp gia tăng độ phì nhiêu đất, kích thích sinh trưởng cây trồng.

The aim of this work was to optimize the growth conditions and continuous production of the enzyme using free and immobilized cells of inulinase by Penicillium funiculosum. The highest yield of enzyme (163.5U/mL) was obtained when the culture was incubated at 27oC and 200 rpm for 96h in a fermentation medium containing both inulin and peptone as sole carbon and nitrogen source, respectively. When the cells of the P. funiculosum were immobilized on different carriers, especially linen fibers, their production ability was successfully maintained for seven successive batches. When the fermentation was carried out using inulin juice prepared from Jerusalem artichoke tubers (in place of pure inulin), inulinase production could be sustained till the second cultivation batch of the P. funiculosum immobilized on linen fibers, yielding 122 U/mL enzyme. Results proved the feasibility of using crude inulin juice as a simple and economic carbon source for the production of inulinase.

Microscopic fungi can be present on a variety of foodstuff, including cheese. They can be responsible for fungal spoilage, causing sensory changes making food unacceptable for human consumption, and posing severe health concerns. Furthermore, some of these organisms are able to resist antimicrobial preservatives provided for by law. Antifungal activity of 15 chemically defined EOs, alone and in mixture, were checked by a microdilution test against isolates of Penicillium funiculosum and Mucor racemosus cultured from rinds of Marzolino, a typical Italian fresh pecorino cheese. Origanum vulgare yielded the lowest MIC values, followed by Salvia sclarea, Ocimum basilicum and Cymbopogon citratus, while Citrus paradisi and Citrus limon were not active. All mixtures showed antifungal activity at lower concentration with respect to MIC values of each EO component, when not in combination. This study is the first to describe the setting up of EOs mixtures to limit spoiling moulds. [ABSTRACT FROM AUTHOR]

Aim. Study of the ability to synthesize extracellular auxins, cytokinins and abscisic acid (ABA) by endophytic and saprotrophic Penicillium funiculosum strains. Methods. Specific biotesting for auxin and cytokinin activities. Quantitative spectrodensitometric thin-layer chromatography. Results. It was shown that the fungal culture filtrates (CF) of P. funiculosum established auxin and cytokinin activities. As a result of study of qualitative and quantitative content of the three classes of phytohormones, was demonstrated that the endophytic strain synthesized high level of physiologically active form of auxins (indole-3-acetic acid) by 55%, cytokinins (zeatin) by 97% and 76% by abscisic acid than saprotrophic one. Conclusions. The studied P. funiculosum strains from different ecotopes synthesized auxins, cytokinins and ABA. The levels and ratio of hormones-stimulants and ABA synthesized by P. funiculosum strains evidence to the possibility of forming of different types of relationships with plants - mutualistic and/or associative. [ABSTRACT FROM AUTHOR]

Aim. Study of the general phytostimulating and gibberellic activities, as well as qualitative and quantitative composition of extracellular gibberellins of endophytic and saprotrophic Penicillium funiculosum strains. Methods. Microbiological, physiological and physico-chemical (HPLC/MS) research methods. Results. It was established that the phytostimulating activity of endophytic P. funiculosum strain was clearly indicated in undiluted culture filtrate (CF) and in 1: 100 ÷ 1: 500 dilutions, whereas for saprotroph it was significantly lower and was observed only at certain dilutions (1:400 and 1:500). In specific gibberellic biotest the CF extract of endophytic strain was three times more active than saprotrophic one - 30% and 10%, respectively. HPLC/MS analysis showed that endophytic and saprotrophic P. funiculosum strains synthesize gibberellins: GA3 (6.8 ng/g dw and 4.1 ng/g dw, respectively), GA4 (4.9 ng/g dw and 11.6 ng/g dw) and trace amounts of GA7. Conclusions. The studied endophytic and saprotrophic P. funiculosum strains were able to synthesize biologically active gibberellins GA3, GA4 and GA7, that may play an important physiological role in the formation of symbiotic and associative relationships with plants. [ABSTRACT FROM AUTHOR]

Aim. Study of endo-, exoglucanase and xylanase activities on the nutrient medium with wheat straw of Penicillium funiculosum strains from different habitats. Methods. The objects of the study were 20 endophytic and soil P. funiculosum strains from culture collection of microscopic fungi of Department of Physiology and Taxonomy of Micromycetes of D.K. Zabolotny Institute of Microbiology and Virology of NASU. Endo-, exoglucanase and xylanase activities were determined quantitative by method with 3.5-dinitrosalicylic acid. Results. Both trophic groups of P. funiculosum were able to synthesize cellulolytic enzyme complex and xylanase. Endo-, exoglucanase and xylanase activities varied at the strain level and were higher for endophytic strains. The maximum values of endo-, exoglucanase and xylanase activities of endophytic strains reached 0.35 ± 0.07, 0.063 ± ± 0.002 and 96.1 ± 2.42 U/ml, respectively. Conclusions. Cellulase and xylanase activities of endophytic and soil P. funiculosum strains were investigated by quantitative methods for the first time. The level of the studied enzymatic activities depended on fungal strain. Endophytic strains characterized by the maximum and minimum cellulolytic and xylanase activities. [ABSTRACT FROM AUTHOR]

ABSTRACT Two induced mutant strains, obtained from the fungus Penicillium funiculosum after UV treatments, were used for protoplast fusion. These mutants (12 and 18) were selected according to their dextranase activity and stability. Seven fusants having dextranase over-yield than their parental strain were obtained. The best dextranase producing fusant (No. 2) was selected for further studies. Dextranase was purified from cell-free culture of fusant 2 by consecutive column chromatography using Q-sepharose FF, Superose 12 prep. grade 60/600 and Mono Q-FPLC. The purification was estimated by SDS-PAGE as well as isoelectric focusing. Four dextranase components (I, II, III and IV) were separated with an estimated molecular weights of 67 kDa. The pI of the all components were found to be around 3.0 as estimated by gel electrophoresis using both broad and low pI calibration protein kits. The dxetranase components showed pH and optimum temperature of 5 and 55oC, respectively. Dextran was the sole carbon source for dextranase production by the dextranase components. The effect of the dextranase components on different types of dextrans were also studied.

The optimization of growth conditions for the production of inulinase by Penicillium funiculosum cells were studied as well as the continuous production of the enzyme using immobilized cells. The highest amount of enzyme (163.5U/mL) was obtained when the producing cells were incubated for 96 hours at 27oC and 200 rpm in a fermentation medium containing both inulin and peptone as sole carbon and nitrogen sources respectively. However, when the cells of the tested microorganism were adsorbed on different carriers, especially linen fibers, their production ability was also successfully maintained, to different extends, for seven successive batches. Moreover, commercially pure inulin is very expensive in only small quantities, this fermentation medium was later substituted by a crude inulin solution obtained from Jerusalem artichoke tubers (Helianthus tuberosus). The crude inulin juice was able to sustain inulinase production during the second batch cultivation of the P. funiculosum, immobilized by their adsorption on linen fibers, in a satisfactory level of about 122U/mL. Moreover, the use of the previously mentioned crude inulin preparation was also compared to the use of either complete or minimal media, composed solely of 1% pure inulin. The method, adopted in this study for inulinase production, is simple, economic, time saving, non-toxic to the microorganism and the loaded linen pads are reusable.

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. [ABSTRACT FROM AUTHOR]

A promising biotechnological strategy in the management of phosphorus (P) fertilization is the use of phosphate-solubilizing fungi to solubilize rock phosphates and allow the recovery of unavailable P fixed to soil particles. Phosphate-solubilizing rhizosphere fungus, Talaromyces funiculosus SLS8, isolated from Neem ( Azadirachta indica) on saline soil, was tolerant to environmental stressors, salinity and agricultural systemic fungicides. Phosphate solubilization under different nutritional conditions was investigated by culturing T. funiculosus SLS8 in Pikovskaya liquid medium containing different nitrogen sources (ammonium sulfate, casein, urea, potassium nitrate or sodium nitrate) and carbon sources (glucose, fructose, galactose or sucrose), NaCl, and three systemic fungicides. The highest concentration of solubilised phosphate (187 mg P L) was achieved after 5 days of incubation in the medium with glucose and ammonium sulphate. The culture pH decreased from 6.5 to 4.2 and HPLC demonstrated organic acid production. Phosphate solubilized was highly negatively correlated with pH (r = −0.96). Increasing salinity had no effect on phosphate solubilization. The maximum tolerance limits to systemic fungicides carbendazim, mancozeb, and hexaconazole were 12.5 μg mL, 2,000 μg mL and 250 μl mL respectively. At these concentrations carbendazim, mancozeb and hexaconazole were found to decrease phosphate solubilization by 55 %, 37 %, and 30 %, respectively. Our results indicate that T. funiculosus SLS8 may be a potential candidate for the development of a biofertilizer for maintaining available phosphate levels in environmentally stressed soils such as saline agricultural soils impacted by systemic fungicide application or seed treatment. [ABSTRACT FROM AUTHOR]

An absorptive fungal strain for uranium adsorption which is named Q5, was isolated from the drainage of a uranium mine in South China. The morphological, physiological and phylogenetic characterizations of the strain Q5 were investigated. The results showed that, the strain Q5 identified as Penicillium funiculosum (99 % similarity in gene sequence). Furthermore, the adsorption performance of P. funiculosum was greatly improved by the mutational method that combined hydroxylamine hydrochloride and UV light. The results showed that the adsorption capacity of the mutated P. funiculosum for U(VI) was obviously better than the non-mutational one at pH 2.0-9.0. [ABSTRACT FROM AUTHOR]

Garlic oil is a kind of fungicide, but little is known about its antifungal effects and mechanism. In this study, the chemical constituents, antifungal activity, and effects of garlic oil were studied with Penicillium funiculosum as a model strain. Results showed that the minimum fungicidal concentrations (MFCs, v/ v) were 0.125 and 0.0313 % in agar medium and broth medium, respectively, suggesting that the garlic oil had a strong antifungal activity. The main ingredients of garlic oil were identified as sulfides, mainly including disulfides (36 %), trisulfides (32 %) and monosulfides (29 %) by gas chromatograph-mass spectrometer (GC/MS), which were estimated as the dominant antifungal factors. The observation results by transmission electron microscope (TEM) and scanning electron microscope (SEM) indicated that garlic oil could firstly penetrate into hyphae cells and even their organelles, and then destroy the cellular structure, finally leading to the leakage of both cytoplasm and macromolecules. Further proteomic analysis displayed garlic oil was able to induce a stimulated or weakened expression of some key proteins for physiological metabolism. Therefore, our study proved that garlic oil can work multiple sites of the hyphae of P. funiculosum to cause their death. The high antifungal effects of garlic oil makes it a broad application prospect in antifungal industries. [ABSTRACT FROM AUTHOR]

Fungal species present in extreme low pH environments are expected to have adapted for tolerance to high H+ concentrations. However, their adaptability mechanism is unclear. In this study, we isolated an acid-tolerant strain of Penicillium funiculosum, which can grow actively at pH 1.0 and thrived in pH 0.6. A major facilitator superfamily transporter (PfMFS) was isolated from an acid-sensitive random insertional mutant (M4) of the fungus. It encodes a putative protein of 551 residues and contains 14 transmembrane-spanning segments. A targeted mutant (M7) carrying an inactivated copy of PfMFS showed an obvious reduction of growth compared with the wild type (WT) and complementation of M7 with PfMFS restored the wild-type level of growth at pH 1.0. Further data showed that the wild-type showed higher intracellular pH than M7 in response to pH 1. Subcellular localization showed that PfMFS was a cell membrane protein. Homology modeling showed structural similarity with an MFS transporter EmrD from Escherichiacoli. These results demonstrate that the PfMFS transporter is involved in the acid resistance and intracellular pH homeostasis of P. funiculosum. [ABSTRACT FROM AUTHOR]

Long time (5 year observation) monitoring results of nine underground power lines (20kV. three-phase. 9 × 3 = 18 cables) are presented. From the experimental data it can be concluded that the cable insulation ageing is a complex process occurring in more successive steps, in which the first and decisive is the external polymeric layer's biodeterioration caused especially by Aspergillus niger and Pénicillium funiculosum. Also be concluded that cables insulation aging can be prevented by cathodic protection of the cables' metallic screen, and already partially aged insulated cables can be rejuvenate to a limited extent. The obtained results validate the proposed model for the degradation mechanism of the buried cables. [ABSTRACT FROM AUTHOR]