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Water-soluble carbohydrates in raw lignocellulosic biomass are converted into hydroxymethylfurfural (HMF) in the acid-based pretreatment, thus increasing the detoxification intensity and reducing the fermentation efficiency of cellulosic ethanol. Therefore, reducing water-soluble carbohydrates in raw corn stover is crucially important to reduce the inhibitors' generation and improve the ethanol fermentation efficiency. In this study, aerobic solid-state fermentation of corn stover by inoculating Aspergillus niger spores converted 83% of the endogenous water-soluble carbohydrates into citric acid, leading to the decrease of 41% of HMF generation and 8% of sulfuric acid usage during the dry acid pretreatment. The reduced inhibitor generation improved the ethanol fermentability by 11% more ethanol than that of the corn stover without water-soluble carbohydrates' removal. This suggests that the removal of the water-soluble carbohydrates before pretreatment significantly reduced the inhibitors' generation in pretreatment and improved the fermentation efficiency of cellulosic ethanol.

Microalgae are photosynthetic eukaryotes which are primary producers in the food chain and also excellent sources for bioactive compounds such as alkaloids, flavonoids, phenols, saponins and other fine chemicals. In the present study, the microalga Scenedesmus bajacalifornicus BBKLP-07 was subjected to soxhlet extraction using solvents like chloroform, acetone, ethanol, methanol and aqueous solvents. All the solvents were tested for the presence of phytochemical constituents such as alkaloids, flavonoids, glycosides, phenols, lignin's, saponins, sterols, tannins, anthraquinone and reducing sugar using the standard procedures. Furthermore, all the crude extracts were subjected to antidiabetic, antioxidant, anti-inflammatory and antimicrobial activities. Antidiabetic activity of the microalgal extracts was observed maximum in Aqueous extract. Methanolic extracts have shown maximum antioxidant activity and chloroform extracts have exhibited highest anti-inflammatory effects. Antimicrobial activities were tested against E.coli, S, typhi, C.perfringens and B.subtilis bacteria and fungi A.niger, and C. albicans. Therefore, the green microalga Scenedesmus bajacalifornicus BBKLP-07 is a rich source of biological active compounds and nutraceuticals and can be exploited for commercial applications.

The high cost of cellulases remains the most significant barrier to the economical production of bio-ethanol from lignocellulosic biomass. The goal of this study was to optimize cellulases and xylanase production by a local indigenous fungus strain (Aspergillus niger DWA8) using agricultural waste (oil palm frond [OPF]) as substrate. The enzyme production profile before optimization indicated that the highest carboxymethyl cellulose (CMCase), filter paper (FPase), and xylanase activities of 1.06 U/g, 2.55 U/g, and 2.93 U/g were obtained on day 5, day 4, and day 5 of fermentation, respectively. Response surface methodology was used to study the effects of several key process parameters in order to optimize cellulase production. Of the five physical and two chemical factors tested, only moisture content of 75% (w/w) and substrate amount of 2.5 g had statistically significant effect on enzymes production. Under optimized conditions of 2.5 g of substrate, 75% (w/w) moisture content, initial medium of pH 4.5, 1 × 10 6 spores/mL of inoculum, and incubation at ambient temperature (±30°C) without additional carbon and nitrogen, the highest CMCase, FPase, and xylanase activities obtained were 2.38 U/g, 2.47 U/g, and 5.23 U/g, respectively. Thus, the optimization process increased CMCase and xylanase production by 124.5 and 78.5%, respectively. Moreover, A. niger DWA8 produced reasonably good cellulase and xylanase titers using OPF as the substrate when compared with previous researcher finding. The enzymes produced by this process could be further use to hydrolyze biomass to generate reducing sugars, which are the feedstock for bioethanol production., (© 2019 American Institute of Chemical Engineers.)

In this study, the ability of the geoactive fungus Aspergillus niger to colonize and transform manganese nodules from the Clarion-Clipperton Zone in both solid and liquid media was investigated. Aspergillus niger was able to colonize and penetrate manganese nodules embedded in solid medium and effect extensive transformation of the mineral in both fragmented and powder forms, precipitating manganese and calcium oxalates. Transformation of manganese nodule powder also occurred in a liquid medium in which A. niger was able to remove the fine particles from suspension which were accumulated within the central region of the resulting mycelial pellets and transformed into manganese oxalate dihydrate (lindbergite) and calcium oxalate dihydrate (weddellite). These findings contribute to an understanding of environmental processes involving insoluble manganese oxides, with practical relevance to chemoorganotrophic mineral bioprocessing applications, and, to the best of our knowledge, represent the first demonstration of fundamental direct and indirect interactions between geoactive fungi and manganese nodules., (© 2019 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd.)

The AraR transcription factor of Aspergillus niger encodes a Zn(II) 2 Cys 6 transcription factor required for the induction of genes encoding arabinolytic enzymes. One of the target genes of AraR is abfA, encoding an arabinofuranosidase. The expression of abfA as well as other L-arabinose-induced genes in A. niger requires the presence of L-arabinose or its derivative L-arabitol as an inducer to activate AraR-dependant gene expression. In this study, mutants were isolated that express L-arabinose-induced genes independently of the presence of an inducer under derepressing conditions. To obtain these mutants, a reporter strain was constructed in a ΔcreA background containing the L-arabinose-responsive promoter (PabfA) fused to the acetamidase (amdS) gene. Spores of the ΔcreA PabfA-amdS reporter strain were UV-mutagenized and mutants were obtained by their ability to grow on acetamide without the presence of inducer. From a total of 164 mutants, 15 mutants were identified to contain transacting mutations resulting in high arabinofuranosidase activity in the medium after growth under non-inducing conditions. Sequencing of the araR gene of the 15 constitutive mutants revealed that 14 mutants carried a mutation in AraR. Some mutations were found more than once and in total nine different point mutations were identified in AraR. The AraR N806I point mutation was reintroduced into a parental strain and confirmed that this point mutation leads to inducer-independent expression of AraR target genes. The inducer independent of L-arabinose-induced genes in the AraR N806I mutant was found to be sensitive to carbon catabolite repression, indicating that the CreA-mediated carbon catabolite repression is dominant over the AraR N806I mutant allele. These mutations in AraR provide new opportunities to improve arabinase production in industrial fungal strains.

The antifungal properties of chitosan-catechin coating and the effect of fruit preservation were studied. We used catechin to modify chitosan to prepare a coating. The purpose of the study was to use chitosan-catechin coating to prolong the preservation time of satsuma oranges. In vitro experiments, the results showed that the antifungal activity of chitosan-catechin increased with increasing concentration, and the results are also significantly effect of comparing to chitosan and catechin alone (*p < 0.05). In vivo studies, chitosan-catechin coating treatment significantly reduced rot caused by Penicillium Citrinum and Aspergillus niger. The physiological and biochemical indexes of the chitosan-catechin coating treatment group were significantly higher than those of the control group (*p ≤ 0.05). In the toxicity test, mice injected with chitosan-catechin solution showed no significant difference compared to the control group. These results indicate that this chitosan-catechin coating may be useful as an antifungal and preserving agent for satsuma oranges. PRACTICAL APPLICATIONS: The fruit after harvest every year is a large loss due to improper storage, and the preservation of fruits is an effective way to reduce losses. The traditional fruit wrap is not degradable, and the preservation effect is relatively general. The chitosan film is a new type of edible fruit wrap, which has the advantages of being edible and easily degradable, and can effectively reduce environmental pollution. Adding catechin to the preparation process of chitosan film can better improve the fresh-keeping effect and prolong the preservation time of the fruit., (© 2019 Wiley Periodicals, Inc.)

Ochratoxin A (OTA) is a mycotoxin produced by filamentous fungi with high impact in food safety due to its toxicity. In the last decade, the presence of OTA was widely reported in different foods. In this study, the ability of Lactobacillus (L.) plantarum CRL 778 to control growth and OTA production by Aspergillus (A.) niger 13D strain, at different water activity (a w ) values (0.955, 0.964, 0.971, 0.982, and 0.995) was determined in vitro. Both parameters were significantly (p<0.05) reduced by the lactobacilli and the effect depended on a w . Greatest growth rate inhibition (46.9%) was obtained at a w =0.995, which is the most suitable value for growth and production of antifungal metabolites (lactic acid, acetic acid, phenyllactic and hydroxyl-phenyllactic acids) by L. plantarum CRL 778. Besides, morphological changes and inhibition of melanin synthesis were observed in colonies of A. niger 13D in presence of L. plantarum CRL 778 at a w ranged between 0.971 and 0.995. In addition, maximum reduction (90%) of OTA production took place at a w =0.971, while inhibition of fungi growth was more evident at a w =0.995. These findings suggest that L. plantarum CRL 778 could be used for control of ochratoxigenic fungal growth and OTA contamination in different fermented foods with a w values between 0.971 and 0.995., (Copyright © 2018 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

Filtering nonwovens that constitute the base material for filtering facepiece respirators (FFRs) used for the protection of the respiratory system against bioaerosols may, in favourable conditions, promote the development of harmful microorganisms. There are no studies looking at the impact that different types of filtering nonwovens have on microorganism survival, which is an important issue for FFR producers and users. Five commercial filtering nonwovens manufactured using diverse textile technologies (i.e., needle-punching, melt-blown, spun-bonding) with different structural parameters and raw material compositions were used within our research. The survival of microorganisms on filtering nonwovens was determined for E. coli , S. aureus , B. subtilis bacteria; C. albicans yeast and A. niger mould. Samples of nonwovens were collected immediately after inoculum application (at 0 h) and after 4, 8, 24, 48, 72, and 96 h of incubation. The tests were carried out in accordance with the AATCC 100-1998 method. Survival depended strongly on microorganism species. E. coli and S. aureus bacteria grew the most on all nonwovens tested. The structural parameters of the nonwovens tested (mass per unit area and thickness) and contact angle did not significantly affect microorganism survival.

Agroforestry industries in the world generate lignocellulosic wastes that can be a huge problem of pollution, or the wastes can be used for different biotechonological applications such as substrates for microorganism growth and enzyme production. Fungi such as Aspergillus niger can grow in almost every substrate and produce hydrolytic enzymes such as endoxylanases, giving added value to agroforestry wastes generated by industries in the northeast of Argentina. In this context, the aim of this work was to use agroforestry wastes as substrates for the production of endoxylanases by Aspergillus niger and to optimize nitrogen sources and physical variables for the highest endoxylanase activity. A. niger LBM 055 and A. niger LBM 134 produced high endoxylanase levels when they were grown with sugarcane and cassava bagasses as carbon sources. A. niger LBM 134 reached the highest endoxylanase activity when nitrogen sources and physical variables were optimized. The fungus exhibited up to 110 U mL -1 of endoxylanase activity when it was grown with sugarcane bagasse and more than 160 U mL -1 with cassava bagasse. Therefore, endoxylanase production was optimized using agricultural bagasses and cost 20 times less than enzyme production using synthetic xylan.

We studied localization and physiological activities of a lectin showing specific binding to N-acetylhexosamines, termed HOL-18, purified from Japanese black sponge (Halichondria okadai). Antiserum against the lectin was generated in rabbit and applied for immunohistochemical analyses. HOL-18 was expressed specifically around water pores and on spicules of sponge tissues. It showed strong binding to a variety of N-acetylhexosamines: N-acetyl D-glucosamine, N-acetyl D-galactosamine, N-acetyl mannosamine, N-acetyl muramic acid, and N-acetyl neuraminic acid. Hemagglutination induced by the lectin was inhibited by lipopolysaccharides and a peptidoglycan. HOL-18 inhibited growth of a gram-positive bacterium (Listeria monocytogenes), gram-negative bacteria (Escherichia coli, Shigella boydii, Pseudomonas aeruginosa), and a fungus (Aspergillus niger). It displayed anti-biofilm activity against P. aeruginosa. HOL-18 was internalized into conidiophores of A. niger, and displayed notable antifungal activity. Fluorescence microscopy revealed binding and incorporation of the lectin into human cancer cell lines HeLa, MCF-7, and T47D, but not Caco-2. HOL-18 displayed dose-dependent cytotoxic effects against HeLa, MCF-7, and T47D, with respective IC 50 values 40, 52, and 63 μg/mL. In HeLa cells, it activated phosphorylation of MAPK pathway molecule (ERK 1/2 ) and activated caspase-3 to trigger apoptosis. HOL-18 thus has the potential to upregulate metabolic pathways in higher animal cells through binding to N-acetylhexosamines., (Copyright © 2018. Published by Elsevier B.V.)

Ochratoxin A (OTA) is a nephrotoxic mycotoxin which may contaminate various foods and feed products worldwide. Aspergillus niger is one of the species responsible for OTA contamination in grapes and derived products. This species has recently been split into A. niger and Aspergillus welwitschiae. Both species can not be distinguished by phenotypic or extrolite profiles and to date there is no ecophysiological information of A. welwitschiae. The aim of this study was to determine the effects of water activity (a w ) (0.90; 0.95 and 0.98-0.99), culture media (Yeast Extract Sucrose Broth (YESB); Synthetic Grape Juice Medium (SGM); White grape juice (WGJ)) and temperature (15 °C, 25 °C and 35 °C) on the growth and OTA production of four strains of A. niger and six strains of A. welwitschiae. The assay was performed in microtiter plates, determining the absorbance at 530 nm and the concentration of OTA at 1, 2, 4 and 10 days. No significant differences were observed in absorbance and OTA values between the two species under study. The highest absorbance values were recorded in YESB, followed by SGM and WGJ. Absorbance values increased with increasing a w and temperature. The highest OTA values were obtained at 0.98-0.99 a w and the best culture media for OTA production was YESB, followed by WGJ and SGM. The studied strains of A. niger produced the highest mean OTA level at 25 °C whereas A. welwitschiae strains produced the highest mean OTA concentration at 15 °C, although not differing significantly from concentration produced at 25 °C. To our knowledge, this is the first report on the impact of some environmental factors on growth and OTA production by A. welwitschiae., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this study, the myco-reduction potential of fungi isolated from soil was ascertained by Norkrans shake flask experiment contaminated with chromium(VI). Fungal tolerance assay and induced tolerance training of the fungi were also carried out. Aspergillus niger, Penicillium, and Saccharomyces strains were isolated from the soil samples using culture based technique. Norkrans samples were collected and analyzed for Cr(VI) concentration using diphenylcarbazide spectrophotometric method. Penicillium strain was observed to be most effect at Cr(VI) concentrations of 16.1 and 8.1 mg L -1 since it was able to reduce Cr(VI) more than Saccharomyces strain and A. niger on day 20. Bio-sorption kinetics for this study was better described by pseudo second order model while Langmuir isotherm model fitted better to the equilibrium data. There was virtually steady increase in fungal growth for all the treatments through-out the experimental period. Significant negative correlation (p < 0.05) was observed between fungal growth and Cr(VI) reduction rate. The results from the induced tolerance training showed that Penicillium had the highest tolerance index (TI) values at 18, 20, and 25 mg L -1 concentrations of Cr(VI) compared to A. niger and Saccharomyces strain. These results demonstrated that these fungi have the potential to bio-absorb Cr(VI) and if properly harnessed, could be used in place of conventional remediation technology to clean-up the Cr(VI) contaminant in the field., (© 2018 The Authors. Journal of Basic Microbiology Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The development of novel solutions to fight microbial food contaminants rests upon two pillars, which are the development of resistant strains and consumers' desire for a reduced consumption of synthetic drugs. Natural antimicrobial peptides possess the qualities to overcome these issues. De novo synthesis of novel antifungal compounds is a major progress that has been facilitated by the identification of parameters involved in the antimicrobial activity. A 14-residue peptide named KK14, with the sequence KKFFRAWWAPRFLK-NH 2 , was designed and inhibited conidial germination and fungal growth of food contaminants within the range 6.25 to 50 μg/ml and 6.25 to 100 μg/ml, respectively. The study of three analogues of the peptide highlighted the role of some residues in the structural conformation of the peptide and its antifungal activity. The substitution of a Pro residue with Arg increased the helical content of the peptide not only its antifungal activity but also its cytotoxicity. The insertion of an unnatural bulky residue β-diphenylalanine or a full d-enantiomerization overall increased the antifungal potency. The four peptides showed similar behaviour towards salt increase, heat treatment, and pH decrease. Interestingly, the denantiomer remained the most active at high pH and after proteolytic digestion. The four peptides did not present haemolytic activity up to 200 μg/ml but had different behaviours of cytotoxicity. These differences could be crucial for potential application as pharmaceutical or food preservatives., (© 2018 European Peptide Society and John Wiley & Sons, Ltd.)

Lanthanum is an important rare earth element and has many applications in modern electronics and catalyst manufacturing. However, there exist several obstacles in the recovery and cycling of this element due to a low average grade in exploitable deposits and low recovery rates by energy-intensive extraction procedures. In this work, a novel method to transform and recover La has been proposed using the geoactive properties of Aspergillus niger. La-containing crystals were formed and collected after A. niger was grown on Czapek-Dox agar medium amended with LaCl 3 . Energy-dispersive X-ray analysis (EDXA) showed the crystals contained C, O, and La; scanning electron microscopy revealed that the crystals were of a tabular structure with terraced surfaces. X-ray diffraction identified the mineral phase of the sample as La 2 (C 2 O 4 ) 3 ·10H 2 O. Thermogravimetric analysis transformed the oxalate crystals into La 2 O 3 with the kinetics of thermal decomposition corresponding well with theoretical calculations. Geochemical modelling further confirmed that the crystals were lanthanum decahydrate and identified optimal conditions for their precipitation. To quantify crystal production, biomass-free fungal culture supernatants were used to precipitate La. The results showed that the precipitated lanthanum decahydrate achieved optimal yields when the concentration of La was above 15 mM and that 100% La was removed from the system at 5 mM La. Our findings provide a new aspect in the biotransformation and biorecovery of rare earth elements from solution using biomass-free fungal culture systems.

The veA gene is a key regulator governing morphogenetic development and secondary metabolism in many fungi. Here, we characterized and disrupted a veA orthologue in an ochratoxigenic Aspergillus niger strain. Morphological development, ochratoxin A (OTA) biosynthesis, and oxidative stress tolerance in the wild-type and veA disruption strains were further analyzed. Accordingly, the link between the veA gene and development of specific gene brlA, OTA biosynthesis key gene pks, and oxidative-stress-tolerance-related gene cat was explored. Results demonstrated that the veA gene acts as a positive regulator of conidia production, OTA biosynthesis, and oxidative stress tolerance in A. niger, regardless of light conditions. Darkness promoted conidial production and OTA biosynthesis in the A. niger wild-type strain. Our results contribute to a better understanding of the veA regulatory mechanism and suggest the veA gene as a potential target for developing control strategies for A. niger infection and OTA biosynthesis.

The remarkable ability of microorganisms to develop resistance to conventional antibiotics is one of the biggest challenges that the pharmaceutical industry currently faces. Recent studies suggest that antimicrobial peptides discovered in spider venoms may be useful resources for the design of structurally new anti-infective agents effective against drug-resistant microorganisms. In this work, we found an anionic antibacterial peptide named U₁-SCRTX-Lg1a in the venom of the spider Loxosceles gaucho . The peptide was purified using high-performance liquid chromatography (HPLC), its antimicrobial activity was tested through liquid growth inhibition assays, and its chemical properties were characterized using mass spectrometry. U₁-SCRTX-Lg1a was found to show a monoisotopic mass of 1695.75 Da, activity against Gram-negative bacteria, a lack of hemolytic effects against human red blood cells, and a lack of cytotoxicity against human cervical carcinoma cells (HeLa). Besides this, the sequence of the peptide exhibited great similarity to specific regions of phospholipases D from different species of Loxosceles spiders, leading to the hypothesis that U₁-SCRTX-Lg1a may have originated from a limited proteolytic cleavage. Our data suggest that U₁-SCRTX-Lg1a is a promising candidate for the development of new antibiotics that could help fight bacterial infections and represents an exciting discovery for Loxosceles spiders.

With increasing demand for novel and potent antimicrobial agents to combat cross-infections and infectious diseases, silver and copper based nanoparticles (NPs) deposited over supports such as montmorillonite (MMT) are playing a crucial role in shaping the current research scenario. Although materials based on Ag NP and Cu NP on MMT have been reported, its toxicological properties on human cell lines have not been accounted for. This paper reports a comparative study on synthesis, antibacterial, antifungal and toxicological behavior of Ag and Cu NPs deposited over MMT nanosheets synthesized by employment of different reduction media. The effect of synthesized NP-MMT hybrids on human erythrocytes and fibroblast cells has been evaluated. The NP formation was facilitated using borohydride and ethyl alcohol (wet chemical route) and photo-reduction and thermal treatment (physical reduction route). The NP-MMT hybrids showed NP formation over supporting silicate layers with particle size ~10-50 nm confirmed by TEM micrographs and loading of ~6-22 wt% of metallic element by EDX analysis. The MMT layers were peeled apart to accommodate NPs inside its galleries, confirmed by increased d-value in powder WAXD. The NP-hybrids showed excellent inhibition zone against bacteria E.coli and S. aureus and fungi A. niger. RBC hemolysis and cytocompatibility assay were performed in vitro to advocate its safety to live human cells. These hybrid materials are potential candidates for new generation advanced antimicrobial materials with less toxicity and highly potent behavior., (Copyright © 2018. Published by Elsevier B.V.)

The present paper aims to elucidate the structural, thermal and mechanical properties of soybean polysaccharide (SSPS)/Cloisite 30B. Tensile strength of the nanocomposite films improved with incorporation of nanoparticles, whereas elongation at break decreased. Surface roughness of the samples increased with the addition of nanoclay. Neat SSPS film and SSPS-1% Cloisite 30B had a relatively smooth surface with no irregularities, while for the samples containing 3 and 7% Cloisite 30B, the surface was rough. DSC analysis demonstrated that following an increase in nanoparticles content, the melting temperature of the nanocomposite elevated, whereas, glass transition temperature decreased. The results of antibacterial activity indicated that Cloisite 30B could inhibit the growth of Salmonella typhi PTCC 1609, Staphylococcus epidermis PTCC 1114 (ATCC 12228) and Listeria monocytogenes PTCC 1165. SSPS-Cloisite 30B nanocomposite could not inhibit the growth of Aspergillus niger. The results demonstrated that the migration of nanoparticles might happen into deionized water as a food simulant, but they could not migrate into bread as a food model. Furthermore, it was found that Cloisite 30B nanoparticles had cytotoxicity effect, and thus, it is recommended that Cloisite 30B/SSPS nanocomposites be used only for the packaging of solids foods such as bread., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The present study compares for the first time the effects of h-MoO3 and α-MoO3 against two fungal strains: Aspergillus niger and Aspergillus flavus. The h-MoO3 nanoparticles were more toxic to both fungi than α-MoO3. The toxic effects of h-MoO3 were more pronounced toward A. flavus, which presented a growth inhibition of 67.4% at 200 mg L-1. The presence of the nanoparticles affected drastically the hyphae morphology by triggering nuclear condensation and compromising the hyphae membrane. Further analysis of the volatile organic compounds (VOCs) produced by both fungi in the presence of the nanomaterials indicated important metabolic changes related to programmed cell death. These nanomaterials induced the production of specific antifungal VOCs, such as β-Elemene and t-Cadinol, by the fungi. The production of essential enzymes involved in fungal metabolism, such as acid phosphatase, naphthol-As-BI-phosphohydrolase, β-galactosidase, β-glucosidase and N-acetyl-β-glucosaminidase, reduced significantly in the presence of the nanomaterials. The changes in enzymatic production and VOCs corroborate the fact that these nanoparticles, especially h-MoO3, exert changes in the fungal metabolism, triggering apoptotic-like cell death responses in these fungi.

Background: Pectinase enzymes present a high priced category of microbial enzymes with many potential applications in various food and oil industries and an estimated market share of $ 41.4 billion by 2020., Results: The production medium was first optimized using a statistical optimization approach to increase pectinase production. A maximal enzyme concentration of 76.35 U/mL (a 2.8-fold increase compared with the initial medium) was produced in a medium composed of (g/L): pectin, 32.22; (NH 4 ) 2 SO 4 , 4.33; K 2 HPO 4 , 1.36; MgSO 4 .5H 2 O, 0.05; KCl, 0.05; and FeSO 4 .5H 2 O, 0.10. The cultivations were then carried out in a 16-L stirred tank bioreactor in both batch and fed-batch modes to improve enzyme production, which is an important step for bioprocess industrialization. Controlling the pH at 5.5 during cultivation yielded a pectinase production of 109.63 U/mL, which was about 10% higher than the uncontrolled pH culture. Furthermore, fed-batch cultivation using sucrose as a feeding substrate with a rate of 2 g/L/h increased the enzyme production up to 450 U/mL after 126 h., Conclusions: Statistical medium optimization improved volumetric pectinase productivity by about 2.8 folds. Scaling-up the production process in 16-L semi-industrial stirred tank bioreactor under controlled pH further enhanced pectinase production by about 4-folds. Finally, bioreactor fed-batch cultivation using constant carbon source feeding increased maximal volumetric enzyme production by about 16.5-folds from the initial starting conditions.

Phenolic compounds are part of the protection system of plants against all kinds of pests and are therefore considered a natural alternative to conventional plant protection agents. Five phenolic compounds (phlorizin, resveratrol, ferulic acid, 5-n-alkylresorcinols, and quercetin) were used as active ingredients in the preparation of a bioactive emulsion and screened for their ability to inhibit the growth of four phytopathogenic fungi - Aspergillus niger, Botrytis cinerea, Penicillium expansum, and Fusarium culmorum. By using design of experiments, several interactions between the phenols were determined, however, ferulic acid was identified as highly effective against the growth of Botrytis cinerea. The growth-inhibitory effect of this emulsion was enhanced by adapting the ferulic acid concentration (0.085%, m/v) with the help of response surface methodology. Subsequent tests confirmed the activity against the growth of nine fungicide-resistant Botrytis cinerea isolates and showed promising results against the infection of injured and contaminated grapes., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

To further understand fermentation kinetics of sodium gluconate (SG) production by Aspergillus niger with different inlet oxygen concentrations, logistic model for cell growth and two-step models for SG production and glucose consumption were established. The results demonstrated that the maximum specific growth rate (µ m ) presented exponential relationship with inlet oxygen concentration and the maximum biomass (X m ) exhibited linear increase. In terms of SG production, two-step model with Luedeking-Piret equation during growth phase and oxygen-dependent equation during stationary phase could well fit the experimental data. Notably, high inlet oxygen concentration exponentially improved SG yield (Y P/S ), whereas biomass yield to glucose (Y X/S ) and cell maintenance coefficient (m) were almost independent on inlet oxygen concentration, indicating that high oxygen supply enhancing SG synthesis not only functioning as a substrate directly, but also regulating glucose metabolism towards SG formation. Finally, the applicability and predictability of the proposed models were further validated by additional experiments.

Development of efficient ocular drug delivery system for antifungal drugs becomes a must nowadays to face and eradicate the widely spread ophthalmic fungal infections. Itraconazole, a triazole antifungal, is struggling to penetrate the cornea and subsequently, its efficacy is limited. The aim of this study was to enhance itraconazole corneal penetration through utilizing the minimum surfactant amount in presence of β-cyclodextrin which acted as a dissolution and permeation enhancer. β-Cyclodextrin consolidated micellar dispersions (CCMD) were prepared after an initial screening to select the composition of surfactant(s). The preparation was done according to a modified melt dispersion technique. The prepared CCMD were characterized through the analysis of their particle size, zeta potential and solubilization efficiency. The optimum formula was chosen based on a factorial response surface analysis and it was composed of 17:1 w/w surfactant/drug, 30:1 w/w cyclodextrin/drug ratios and 0.02% polyethylene oxide. This formula was subjected to in vitro characterization including release, imaging by transmission electron microscope, mucoadhesion, stability, in addition to the determination of the minimum inhibitory concentration. Moreover, the ex vivo/in vivo permeation, safety and efficacy profiles were determined. The optimized CCMD formula was found to be significantly safe, stable, mucoadhesive and efficient to permeate the drug through rabbits' corneas. Consequently, the optimized CCMD formulation can be a promising, safe and efficient platform for the transcorneal delivery of lipophilic drugs including most antifungals., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Coffee husk (CH), a waste obtained from processing of coffee cherries via dry method, causes serious environmental problems. In this study, strategies were designed to utilize CH for succinic acid (SA) production. Three different CH hydrolysis methods: thermal, thermochemical and crude enzymes obtained by solid state fermentation of Aspergillus niger and Trichoderma reesei, were evaluated to generate fermentable feedstock for SA production using Actinobacillus succinogenes. The feasibility of these pretreatment methods was investigated. Accordingly, thermochemical hydrolysis using H 2 SO 4 at 121 °C for 30 min, appeared the most effective method for CH hydrolysis, producing 24.4 g/L of reducing sugars (RS). Finally, 19.3 g/L of SA with yield and productivity of 0.95 g SA/g RS and 0.54 g/L/h, respectively, were obtained using CH hydrolysate. The current study revealed an alternative way of utilization coffee waste for value addition while mitigating environmental problems caused by its disposal.

This study aims at identifying the populations of filamentous fungi present on a Brazilian contemporary painting, whose conservation status was compromised and showed evident signs of deterioration by microbial action. In addition, to correlate the biodeterioration potential of the isolated fungal strains, cellulolytic activity testing was performed and, finally, the biocide treatment against microbial growth was carried out. A total of nine isolates of filamentous fungi were detected and three distinct taxa were identified: Aspergillus flavus, Aspergillus niger and Penicillium citrinum. The detection of the enzymatic activity of the isolates by cellulolytic plate assay revealed the potential of filamentous fungal species in causing the deterioration of paintings. Our results showed that the presence of each strain of filamentous fungi correlated with the distribution of the paint colour, suggesting a tropism of certain species for specific dyes used. In addition, strains of A. niger showed a lower enzymatic activity index, despite the aesthetic damage that this fungal specie caused on the artwork. In conclusion, this study demonstrated that the identification of the microbiota obtained from the painting may help contrasting its biodeterioration and it described a successful antifungal treatment on a contemporary art piece. Contemporary art in Brazil arose from postmodernism, through the artistic manifestations included in the Neoconcrete Manifest of 1959, as well as with the use of innovative techniques. It was a consequence of the interaction between concept and language, displayed by the artists on a set of unconventional materials and objects. Despite being a 20th century painting, the concern of the artist to keep the art object in a good conservation state has driven the realization of this work and these microbial prospecting studies, associated with artwork deterioration, may contribute to the preservation of the Brazilian contemporary heritage., (© 2018 The Society for Applied Microbiology.)

The coordination properties of 2-picolinehydroxamic acid towards cobalt(II) in aqueous solution were determined by a pH-metric method and confirmed by spectroscopic (UV-Vis and ESI-MS) studies. The results show the formation of mononuclear complexes, as well as of metallacrowns (MC). All methods indicate a high tendency of 2-picolinehydroxamic acid to form cobalt(II) metallacrown 12-MC-4. ESI-MS additionally confirms 15-MC-5 and 18-MC-6, stabilized by a sodium ion and methanol. The complexes observed in the speciation model at a pH about 7.2 were studied for their DNA-binding ability. The decrease of absorbance in the range of ca 310-400 nm indicates effective binding to calf thymus DNA by 2-picolinehydroxamic acid complexes, via intercalative mode. The antimicrobial properties of 2-picolinehydroxamic acid, cobalt(II) ions and of the complexes formed in the Co(II) - ligand system were determined against Gram-positive bacteria (Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Bacillus subtilis), Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli, Helicobacter pylori) and fungal strains (Candida, Aspergillus niger). The results indicate that the complexes demonstrate greater antibacterial and antifungal activity for most strains than the ligand. Both the complexes and the ligand induce a slight decrease in the metabolic activity of cells, while the complexes do not damage the cell nuclei. The 2-picolinehydroxamic acid complexes activate the human monocytic cells, suggesting they have immunomodulating properties, which are particularly important in combating infections caused by strains resistant to other drugs., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Itaconic acid is a five-carbon dicarboxylic acid with an unsaturated alkene bond, frequently used as a building block for the industrial production of a variety of synthetic polymers. It is also one of the major products of fungal "overflow metabolism" which can be produced in submerged fermentations of the filamentous fungus Aspergillus terreus. At the present, molar yields of itaconate are lower than those obtained in citric acid production in Aspergillus niger. Here, we have studied the possibility that the yield may be limited by the oxygen supply during fermentation and hence tested the effect of the dissolved oxygen concentration on the itaconic acid formation rate and yield in lab-scale bioreactors. The data show that a dissolved oxygen concentration of 2% saturation was sufficient for maximal biomass formation. Raising it to 30% saturation had no effect on biomass formation or the growth rate, but the itaconate yield augmented substantially from 0.53 to 0.85 mol itaconate/mol glucose. Furthermore, the volumetric and specific rates of itaconic acid formation ameliorated by as much as 150% concurrent with faster glucose consumption, shortening the fermentation time by 48 h. Further increasing the dissolved oxygen concentration over 30% saturation had no effect. Moreover, we show that this increase in itaconic acid production coincides with an increase in alternative respiration, circumventing the formation of surplus ATP by the cytochrome electron transport chain, as well as with increased levels of alternative oxidase transcript. We conclude that high(er) itaconic acid accumulation requires a dissolved oxygen concentration that is much higher than that needed for maximal biomass formation, and postulate that the induction of alternative respiration allows the necessary NADH reoxidation ratio without surplus ATP production to increase the glucose consumption and the flux through overflow metabolism.

Effects of Aspergillus niger was investigated on the strength of cotton textile specimens impregnated with nano-wollastonite, and then compared with normal specimens. Cotton strips were cut and prepared in warp and wept directions according to the standard specifications ASTM D-5035. Results showed that incubation of A. niger on specimens for three months resulted in a significant decrease in tensile stress as well as weight mass change in both directions. Impregnating specimens with NW ameliorated the negative effects of fungal attack on tensile stress to a considerable extent. Moreover, weight change was significantly decreased. It is concluded that NW positively protect cotton textile against A. nigra; the ultimate NW-content should be studied in complimentary studies.

Here, the authors describe a simple method to formulate the nanodispersion of hexaconazole (hexa); henceforth, referred to as nanohexaconazole (N-hexa) that is water soluble and effective against several species of Aspergillus . Size and shape of the prepared nanocomposite was determined with high-resolution transmission electron microscopy and field-emission scanning electron microscopy. Nanohexaconazole structure was further confirmed by Fourier-transform infrared spectroscopy and gas chromatography-mass spectrometry. The antifungal efficacy of nanohexaconazole (N-hexa) was studied in vitro , compared with micronised hexaconazole (M-hexa) at different doses (5 ppm, 10 ppm and control) against two food pathogenic fungi: Aspergillus niger (MTCC 282, MTCC 2196 and BDS 113) and Aspergillus fumigatus through poisoned food technique. A dose-dependent significant growth inhibition was observed in nanohexaconazole (N-hexa) treated fungal sample compared with that of micronised hexaconazole (M-hexa). Micrographic studies for the morphological analysis of control and nanohexaconazole (N-hexa) treated fungal samples were done, exhibited an alternation in fungal morphology. Results showed that nanohexaconazole (N-hexa) is more efficacious than commercially available micronised hexaconazole (M-hexa). In future nanohexaconazole (N-hexa) could be a possible candidate for modern medical science and also reduce damage to the environment from injudicious use of pesticides.

Plant defensins are small, cysteine-rich antimicrobial peptides of the immune system found in several organs during plant development. A synthetic peptide, KT43C, a linear analogue of the native Cp-thionin II found in cowpea seeds, was evaluated for its antifungal potential. It was found that KT43C displayed antifungal activity against Fusarium culmorum, Penicillium expansum and Aspergillus niger. Like native plant defensins, KT43C showed thermostability up to 100 °C and cation sensitivity. The synthetic peptide decreased the fungal growth without inducing morphogenic changes in the fungal hyphae. Non-inhibitory concentrations of the peptide induced permeabilization of the fungal membrane. In addition, high concentrations of KT43C induced the production of reactive oxygen species in the granulated cytoplasm. To investigate potential applications, the peptide was used as an additive in the preparation of dough which did not contain yeast. This peptide delayed the development of fungal growth in the dough by 2 days. Furthermore, KT43C did not induce red blood cell lysis up to a concentration of 200 μg.ml -1 . These results highlight the potential for the use of synthetic antimicrobial defensins for shelf-life extension of food products., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

DKxanthenes are a class of yellow secondary metabolites produced by myxobacterial genera Myxococcus and Stigmatella . We identified a putative 49.5 kb DKxanthene biosynthetic gene cluster from Myxococcus stipitatus DSM 14675 by genomic sequence and mutational analysis. The cluster was comprisedof 15 genes (MYSTI&#95;06004-MYSTI&#95;06018) encoding polyketide synthases, non-ribosomal peptide synthases, and proteins with unknown functions. Disruption of the genes by plasmid insertion resulted in defects in the production of yellow pigments. High-performance liquid chromatography and liquid chromatography-tandem mass spectrometry analysis indicated that the yellow pigments produced by M. stipitatus DSM 14675 might be noble DKxanthene derivatives. M. stipitatus did not require DKxanthenes for the formation of heat-resistant viable spores, unlike Myxococcus xanthus . Furthermore, DKxanthenes showed growth inhibitory activity against the fungi Aspergillus niger , Candida albicans , and Rhizopus stolonifer .

The impact of formulation and desiccation on the shelf life of phosphate (P)-solubilising microorganisms is often under-studied, particularly relating to their ability to recover P-solubilisation activity. Here, Penicilllium bilaiae and Aspergillus niger were formulated on vermiculite (V) alone, or with the addition of protectants (skimmed milk (V + SM) and trehalose (V + T)), and on sewage sludge ash with (A + N) and without nutrients (A), and dried in a convective air dryer. After drying, the spore viability of P. bilaiae was greater than that of A. niger. V formulations achieved the highest survival rates without being improved by the addition of protectants. P. bilaiae formulated on V was selected for desiccation in a fluidised bed dryer, in which several temperatures and final water activities (aw) were tested. The highest spore viability was achieved when the formulation was dried at 25 °C to a final aw >0.3. During three months' storage, convective air dried formulations were stable for both strains, except in the presence of skimmed milk for P. bilaiae which saw a decrease in spore viability. In the fluidised bed-dried formulations, when aw >0.3, the loss in viability was higher, especially when stored at 20 °C, than at aw <0.1. P-solubilisation activity performed on ash was preserved in most of the formulations after desiccation and storage. Overall, a low drying temperature and high final aw positively affected P. bilaiae viability, however a trade-off between higher viability after desiccation and shelf life should be considered. Further research is needed to optimise viability over time and on more sustainable carriers., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Efficient hydrolysis of holocellulose depends on a proper balance between cellulase (endoglucanase, exoglucanase, β-glucosidase) and xylanase activities. The present study aimed to induce the production of cellulases and xylanases using liquid cultures (one, two, three, and four fungal strains on the same bioreactor) of wild strains of Trichoderma harzianum, Aspergillus niger, and Fusarium oxysporum. The strains were identified by amplification and analysis of the ITS rDNA region and the obtained sequences were deposited in Genbank. Enzymes (endoglucanase, exoglucansae, β-glucosidase, and xylanase activities) and the profile of extracellular protein isoforms (SDS-PAGE) produced by different fungal combinations (N = 14) were analyzed by Pearson's correlation matrix and principal component analysis (PCA). According to our results, induction of endoglucanase (19.02%) and β-glucosidase (6.35%) were obtained after 4 days when A. niger and F. oxysporum were cocultured. The combination of A. niger-T. harzianum produced higher endoglucanase in a shorter time than monocultures. On the contrary, when more than two strains were cultured in the same reactor, the relationships of competition were established, trending to diminish the amount of enzymes and the extracellular protein isoforms produced. The xylanase production was sensible to stress produced by mixed cultures, decreasing their activity. This is important when the aim is to produce cellulase-free xylanase. In addition, exoglucanase activity did not change in the combinations tested.

Background: Pu-erh tea is a traditional Chinese tea and produced by natural solid-state fermentation. Several studies show that the natural microbiota influence caffeine level in pu-erh tea. Our previous research also found that the caffeine declined significantly (p < 0.05) in the fermentation, which suggested that the caffeine level could be influenced by specific strains. The purpose of this study was to isolate and identify microorganisms for caffeine degradation, and this research explored the degradation products from caffeine and optimal condition for caffeine degradation., Results: 11 Fungi were isolated from pu-erh tea fermentation and 7 strains could survive in caffeine solid medium. Two superior strains were identified as Aspergillus niger NCBT110A and Aspergillus sydowii NRRL250 by molecular identification. In the substrate tests with caffeine, A. niger NCBT110A could use caffeine as a potential carbon source while glucose is absent, A. sydowii NRRL250 could degrade 600 mg/L caffeine completely in a liquid medium. During the degradation product analysis of A. sydowii NRRL250, theophylline and 3-methlxanthine were detected, and the level of theophylline and 3-methlxanthine increased significantly (p < 0.05) with the degradation of caffeine. The single factor analysis showed that the optimum conditions of caffeine degradation were 1) substrate concentration of 1200 mg/L, 2) reaction temperature at 30 °C, and 3) pH of 6. In the submerged fermentation of tea infusion by A. sydowii NRRL250, 985.1 mg/L of caffeine was degraded, and 501.2 mg/L of theophylline was produced., Conclusions: Results from this research indicate that Aspergillus sydowii NRRL250 was an effective strain to degrade caffeine. And theophylline and 3-methlxanthine were the main caffeine degradation products.

This study systematically investigated the degradation kinetics and changes in the composition of phenolic compounds in Spanish-style Chalkidiki green olive processing wastewaters (TOPWs) during treatment using Aspergillus niger B60. The fungal growth and phenol degradation kinetics were described sufficiently by the Logistic and Edward models, respectively. The maximum specific growth rate (2.626 1/d) and the maximum degradation rate (0.690 1/h) were observed at 1500 mg/L of total polar phenols, indicating the applicability of the process in TOPWs with a high concentration of phenolic compounds. Hydroxytyrosol and the other simple phenols were depleted after 3-8 days. The newly formed secoiridoid derivatives identified by HPLC-DAD-FLD and LC-MS are likely produced by oleoside and oleuropein aglycon via the action of fungal β-glucosidase and esterase. The treated streams were found to be less phytotoxic with reduced chemical oxygen demand by up to 76%. Findings will provide useful information for the subsequent treatment of residual contaminants.

Currently, there is an absolute concern for all nations in agricultural productivity to meet growing demands of human population. In recent time, biosurfactants produced by diverse group of microorganisms are used to achieve such demands as it is known for its ecofriendly use in elimination of plant pathogens and for increasing the bioavailability of nutrients for plants. Endophytic fungi are the important source of secondary metabolites and novel bioactive compounds for different biological applications. In the present study, endophytic fungi Xylaria regalis (X. regalis) recovered from the cones of Thuja plicata was evaluated for its biosurfactant producing ability and plant growth-promoting abilities through various screening methods and also via its antagonistic activity against phytopathogens like Fusarium oxysporum and Aspergillus niger . In addition, X. regalis was also tested in vivo for a various range of growth parameters in chilli under greenhouse conditions. Significant increase in shoot and root length, dry matter production of shoot and root, chlorophyll, nitrogen, and phosphorus contents of chilli seedlings was found, which reveals its ability to improve the growth of crop plants. Hence, this study suggests the possibility of biosurfactant producing endophytic fungi X. regalis as a source of novel green biosurfactant for sustainable agriculture to achieve growing demands.

Background: Mung bean is a rich source of protein, carbohydrates and fiber content. It also exhibits a high level of antioxidant activity due to the presence of phenolic compounds. Aspergillus flavus and A. niger are the two major fungal strains associated with stored mung bean that lead to post-harvest losses of grains and also cause serious health risks to human beings. Thus there is a need to explore an economical decontamination method that can be used without affecting the biochemical parameters of grains., Results: It was observed that infrared (IR) treatment of mung bean surface up to 70 °C for 5 min at an intensity of 0.299 kW m -2 led to complete visible inhibition of fungal growth. Scanning electron microscopy revealed that surface irregularities and physical disruption of spores coat are the major reasons behind the inactivation of IR-treated fungal spores. It was also reported that IR treatment up to 70 °C for 5 min does not cause any negative impact on the biochemical and physical properties of mung bean., Conclusion: From the results of the present study, it was concluded that IR treatment at 70 °C for 5 min using an IR source having an intensity of 0.299 kW m -2 can be successfully used as a method of fungal decontamination. The fungal spore population was reduced (approximately 5.3 log 10 CFU g -1 reductions) without significantly altering the biochemical and physical properties of grains. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

The efficient use of renewable lignocellulosic feedstocks to obtain biofuels and other bioproducts is a key requirement for a sustainable biobased economy. This requires novel and effective strategies to reduce the cost contribution of the cellulolytic enzymatic cocktails needed to convert the carbohydrates into simple sugars, in order to make large-scale commercial processes economically competitive. Here, we propose the use of the whole solid-state fermentation (SSF) medium of mixed filamentous fungi as an integrated one-pot strategy for on-site enzyme production, biomass hydrolysis, and ethanol production. Ten different individual and mixed cultivations of commonly used industrial filamentous fungi (Aspergillus niger, Aspergillus oryzae, Trichoderma harzianum, and Trichoderma reesei) were performed under SSF and the whole media (without the extraction step) were used in the hydrolysis of pretreated sugarcane bagasse. The cocultivation of T. reesei with A. oryzae increased the amount of glucose released by around 50%, compared with individual cultivations. The release of glucose and reducing sugars achieved using the whole SSF medium was around 3-fold higher than obtained with the enzyme extract. The addition of soybean protein (0.5% w/w) during the hydrolysis reaction further significantly improved the saccharification performance by blocking the lignin and avoiding unproductive adsorption of enzymes. The results of the alcoholic fermentation validated the overall integrated process, with a volumetric ethanol productivity of 4.77 g/L.h, representing 83.5% of the theoretical yield. These findings demonstrate the feasibility of the proposed one-pot integrated strategy using the whole SSF medium of mixed filamentous fungi for on-site enzymes production, biomass hydrolysis, and ethanol production. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:671-680, 2018., (© 2018 American Institute of Chemical Engineers.)

In nature, the fungus Aspergillus niger degrades plant biomass polysaccharides to monomeric sugars, transports them into its cells, and uses catabolic pathways to convert them into biochemical building blocks and energy. We show that when grown in liquid cultures, A. niger takes up plant-biomass derived sugars in a largely sequential manner. Interestingly, this sequential uptake was not mediated by the fungal general carbon catabolite repressor protein CreA. Furthermore, transcriptome analysis strongly indicated that the preferential use of the monomeric sugars is arranged at the level of transport, but it is not reflected in transcriptional regulation of sugar catabolism. Therefore, the results indicate that the regulation of sugar transport and catabolism are separate processes in A. niger.

This contribution investigates aluminium mobilization from main aluminium pools in soils, phyllosilicates and oxyhydroxides, by acidic and chelating exometabolites of common soil fungi Aspergillus niger and A. clavatus. Their exometabolites' acidity as well as their ability to extract aluminium from solid mineral phases differed significantly during incubation. While both strains are able to mobilize aluminium from boehmite and aluminium oxide mixture to some extent, A. clavatus struggles to mobilize any aluminium from gibbsite. Furthermore, passive and active fungal uptake of aluminium enhances its mobilization from boehmite, especially in later growth phase, with strong linear correlation between aluminium bioaccumulated fraction and increasing culture medium pH. We also provide data on concentrations of oxalate, citrate and gluconate which are synthesized by A. niger and contribute to aluminium mobilization. Compared to boehmite-free treatment, fungus reduces oxalate production significantly in boehmite presence to restrict aluminium extraction efficiency. However, in presence of high phyllosilicates' dosages, aluminium is released to an extent that acetate and citrate is overproduced by fungus. Our results also highlight fungal capability to significantly enhance iron and silicon mobility as these elements are extracted from mineral lattice of phyllosilicates by fungal exometabolites alongside aluminium., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Aspergillus niger forms conidia that contain melanin in their cell wall. This black pigment has been shown to protect fungi against UV radiation, and experimental evidence has indicated that it also protects against drought and high salt concentrations. In this study, growth of A. niger was evaluated at low water activity (a w ) and after changes in relative humidity (RH). In addition, deletion strains of A. niger affected in the melanin synthesis pathway were compared. Germination of conidia of the wild-type and deletion strains was observed at 0·81 a w and germ tubes continued growth at a w  ≥ 0·83. Conidia and microcolonies of the different strains were incubated for 1 week at lowered RH (33-84%). Conidia of all strains germinated and formed colonies after exposure to RH ≥33% when transferred back to malt extract medium at a w 0·98. Conidia germinated and showed limited growth at 84% RH. Microcolonies of all strains did not survive an incubation of 1 week at RH ≤75%, but continued growth after exposure to 84% RH. Together, this is the first genetic evidence that melanin does not play a role during germination and radial extension of fungi at low water conditions., Significance and Impact of the Study: Aspergillus niger, a cosmopolitan fungus with melanized conidia, is used here as a model system for fungal growth at low water activity (a w ) and humidity dynamics. From this study it becomes clear that melanin, contrary to what has been suggested before, is not a key factor in survival and growth during situations that mimic indoor conditions. Indoor fungal growth can lead to cosmetic damage to building materials and health problems. This knowledge makes clear that novel ways to limit indoor fungal growth have to be based on interference with other cellular traits of fungi., (© 2018 The Society for Applied Microbiology.)

Oxalic acid has potent nematicidal activity against the root-knot nematode Meloidogyne incognita. In this study, fermentation parameters for oxalic acid production in submerged culture of Aspergillus niger F22 at 23, 25, and 30 °C were optimized in 5-L jar fermenters. The viscosity of the culture broth increased with increasing temperature. There was a negative correlation between oxalic acid production and the apparent viscosity; high volumetric productivity of oxalic acid was obtained at low apparent viscosity (less than 1000 cP), with a productivity of more than 100 mg/L h. When the apparent viscosity was over 2500 cP, the volumetric productivity decreased below 50 mg/L h. In addition, the volumetric mass transfer coefficient, K L a, positively correlated with volumetric productivity. When the K L a value increased from 0.0 to 0.017 /s, the volumetric productivity proportionally increased up to 176 mg/L h. When the temperature decreased, K L a increased due to the decrease in viscosity, leading to increased volumetric productivity. The highest productivity of 7453.3 mg/L was obtained at the lowest temperature, i.e., 23 °C. The nematicidal activity of culture filtrate was proportional to the content of oxalic acid. Based on a constant impeller tip speed, oxalic acid production was successfully scaled up to a 500-L pilot vessel, producing a final concentration comparable to that in the 5-L jar.

Forty Bacillus isolates obtained from maari (used as condiment in Burkina Faso) including 17 B. subtilis, 4 B. circulans, 7 B. pumilus and 6 B. licheniformis were investigated for use as starter cultures in maari production. The isolates were screened by PCR for the sfp gene responsible for the production of the lipopeptide biosurfactant, surfactin. The sfp gene was detected in all of the seventeen B. subtilis isolates, in 2 out of 7 B. pumilus, in 4 out of 6 B. licheniformis whereas no B. circulans was positive for the sfp gene by PCR screening. Furthermore, all the 40 Bacillus spp. were screened for biosurfactant production and inhibitory activity against Aspergillus flavus, A. niger, A. versicolor and Rhizopus oryzae. Results demonstrated a relationship between the presence of the sfp gene and the antifungal activity and biosurfactant production of Bacillus isolates. In addition, molecular typing of the 17 B. subtilis isolates by Multilocus Sequence Typing (MLST) resulted in 15 Sequence Types, one of them included three strains. Randomly Amplified Polymorphic DNA-PCR (RAPD-PCR), used for B. licheniformis, B. megaterium, B. circulans and B. pumilus revealed that the inhibitory activity and biosurfactant production were strain-dependent. Finally, the detection of chitinase (chi) and β-glucanase (glu) biosynthesis genes was found to be associated with the antifungal activity for 16 B. subtilis isolates. The present work provides a greater understanding of the antifungal activity and biosurfactant production ability within the Bacillus spp. isolated from maari and contributes to the selection of Bacillus isolates to be used as starter cultures for controlled production of maari., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Byproducts of food processing can be utilized for the production of high-value-added enzyme cocktails. In this study, we utilized integrated functional omics technology to analyze composition and functional characteristics of extracellular enzymes produced by Aspergillus niger grown on food processing byproducts. The results showed that oligosaccharides constituted by arabinose, xylose, and glucose in wheat bran were able to efficiently induce the production of extracellular enzymes of A. niger. Compared with other substrates, wheat bran was more effective at inducing the secretion of β-glucosidases from GH1 and GH3 families, as well as >50% of proteases from A1-family aspartic proteases. Compared with proteins induced by single wheat bran or soybean dregs, the protein yield induced by their mixture was doubled, and the time required to reach peak enzyme activity was shortened by 25%. This study provided a technical platform for the complex formulation of various substrates and functional analysis of extracellular enzymes.

In its natural environment, the filamentous fungus Aspergillus niger grows on decaying fruits and plant material, thereby enzymatically degrading the lignocellulosic constituents (lignin, cellulose, hemicellulose, and pectin) into a mixture of mono- and oligosaccharides. To investigate the kinetics and stoichiometry of growth of this fungus on lignocellulosic sugars, we carried out batch cultivations on six representative monosaccharides (glucose, xylose, mannose, rhamnose, arabinose, and galacturonic acid) and a mixture of these. Growth on these substrates was characterized in terms of biomass yields, oxygen/biomass ratios, and specific conversion rates. Interestingly, in combination, some of the carbon sources were consumed simultaneously and some sequentially. With a previously developed protocol, a sequential chemostat cultivation experiment was performed on a feed mixture of the six substrates. We found that the uptake of glucose, xylose, and mannose could be described with a Michaelis-Menten-type kinetics; however, these carbon sources seem to be competing for the same transport systems, while the uptake of arabinose, galacturonic acid, and rhamnose appeared to be repressed by the presence of other substrates.

Two new compounds [1-2] were purified from ethyl acetate fraction of Quercus incana. The structure of these compounds is mainly established by using advanced spectroscopic technique such as UV, IR, one-dimensional (ID) and two-dimensional (2D) NMR techniques, and EI mass. The structural formula was deduced to be 4-hydroxydecanoic acid [1] and 4-hydroxy-3-(hydroxymethyl) pentanoic acid [2]. Both isolated compounds were tested for their antimicrobial potential and showed promising antifungal activity against Aspergillus niger and Aspergillus flavus .

The design of novel efficient antimicrobial peptides (AMPs) faces several issues, such as cost of synthesis, proteolytic stability or cytotoxicity. The identification of key determinants involved in the activity of AMPs, such as cationicity and amphipathicity, allowed the synthesis of short peptides with optimized properties. An ultrashort peptide made of the sequence H-Orn-Orn-Trp-Trp-NH2 (O3TR) showed antifungal activity against several contaminants from food products. This peptide inhibited the growth of the filamentous fungi Fusarium culmorum, Penicillium expansum and Aspergillus niger within a range of concentration of 12.5-50μg/ml. In addition, O3TR inhibited the growth of the yeast Saccharomyces cerevisiae, Zygosaccharomyces bailii, Zygosaccharomyces rouxii, Debaryomyces hansenii and Kluyveromyces lactis within the range 12.5-50μg/ml. A derivative peptide, called C12O3TR, made by the addition of lauric acid at the N-terminus of O3TR was 2- to 8-fold more active than O3TR against every species. In addition to the inhibition of conidial germination, O3TR and C12O3TR killed F. culmorum hyphae at 100 and 50μg/ml respectively. The MIC of the two peptides against F. culmorum and Z. bailii after heat treatment at 100°C for 60 min and within the pH range 3-10, were not changed. However, the activity of O3TR against F.culmorum and Z. bailii was strongly reduced in salt solutions, whereas the lauric acid peptide kept its antifungal activity and resistance to proteolytic digestion. The conjugation with lauric acid reduced the random coiled structure and increased the α-helical content of O3TR. After conjugation with the dye tetramethylrhodamine (TMR), both peptides entered F. culmorum spores. They also both induced permeabilization of F. culmorum hyphae but only C12O3TR permeabilized Z. bailii membrane. In contrast to the lipopeptide, O3TR did not show haemolytic or cytotoxic activity when applied at the concentrations that exhibited antifungal potency. The two peptides were challenged against a yeast cocktail of S. cerevisiae and Z. bailii, and A. niger in different commercial beverages. After 7 days, O3TR was able to inhibit the yeast cocktail in a commercial lager and carbonated drink. Due to its antifungal potency, high stability and low cytotoxicity, the tetrapeptide could represent a promising starting point of a novel food preservative., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Cellulases and other enzymes are needed for saccharification of plant biomass in the biorefinery industry. Expression, characterization, and eventual large-scale production of known and novel cellulases requires the ability to express and secrete heterologous enzymes in relevant protein production platforms like Aspergillus niger. A method for cloning and expression of genes for these desirable enzymes in A. niger is presented in this Chapter.

Strategies for achieving global food security include identification of alternative feedstock for use as animal feed, to contribute towards efforts at increasing livestock farming. The presence of theobromine in cocoa pod husks, a major agro-waste in cocoa-producing countries, hinders its utilisation for this purpose. Cheap treatment of cocoa pod husks to remove theobromine would allow largescale beneficial use of the millions of metric tonnes generated annually. The aim of this study was to isolate theobromine-degrading filamentous fungi that could serve as bioremediation agents for detheobromination of cocoa pod husks. Filamentous fungi were screened for ability to degrade theobromine. The most promising isolates were characterized with respect to optimal environmental conditions for theobromine degradation. Secretion of theobromine-degrading enzymes by the isolates was investigated. Theobromine degradation was monitored by HPLC. Of fourteen theobromine-degrading isolates collected and identified by rDNA 5.8S and ITS sequences, seven belonged to Aspergillus spp. and six were Talaromyces spp. Based on the extent of theobromine utilization, four isolates; Aspergillus niger, Talaromyces verruculosus and two Talaromyces marneffei, showed the best potential for use as bioagents for detheobromination. First-time evidence was found of the use of xanthine oxidase and theobromine oxidase in degradation of a methylxanthine by fungal isolates. Metabolism of theobromine involved initial demethylation at position 7 to form 3-methylxanthine, or initial oxidation at position 8 to form 3,7-dimethyuric acid. All four isolates degraded theobromine beyond uric acid. The data suggest that the four isolates can be applied to substrates, such as cocoa pod husks, for elimination of theobromine., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Background: The benzimidazole ring is an important pharmacophore in modern drug discovery. Mannich reaction is one of the versatile reaction widely used in organic synthesis. Mannich base derivatives play an important role in medical field with diverse biological actions., Objective: A series of N-(benzimidazol-1-ylmethyl)-4-chlorobenzamide derivatives (3a- 3m) were synthesized and evaluated for anti-inflammatory and antimicrobial potential., Method: Mannich reaction was used to synthesize N-(benzimidazol-1-ylmethyl)-4- chlorobenzamide analogues. The structures of novel target compounds were elucidated by spectral and analytical techniques and screened for in vivo anti-inflammatory activity and ulcerogenic activity. In addition, the prepared derivatives were also evaluated for in vitro antimicrobial activity against gram negative, gram positive and fungal strains. Further, in silico studies were carried out to define the interaction of the title compounds with COX-2 enzyme and microbial protein., Results: The results revealed that out of thirteen molecules, compound 3a (containing chloromethyl substituent at 2-position of benzimidazole) showed significant antiinflammatory effect at a dose of 100 mg/kg p.o. and the experimental data was statistically significant at p≤0.05 level. Diclofenac sodium was taken as standard drug for antiinflammatory activity. Furthermore, derivative 3e (containing 2-chlorophenyl moiety at 2- position of benzimidazole scaffold) was found to be the most effective antimicrobial compound among the synthesized derivatives. Ciprofloxacin and clotrimazole were used as reference antimicrobial agents. Results from in vivo and in vitro studies of synthesized analogues were found to be in good correlation with in silico study., Conclusion: These results designate that N-(Benzimidazol-1-ylmethyl)-4-chlorobenzamide analogues, substituted with halogen functionality, could be used as potential lead for designing more potent anti-inflammatory and antimicrobial agents., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)