Showing papers on "Aspergillus niger published in 2007"

Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88

Abstract: The filamentous fungus Aspergillus niger is widely exploited by the fermentation industry for the production of enzymes and organic acids, particularly citric acid. We sequenced the 33.9-megabase genome of A. niger CBS 513.88, the ancestor of currently used enzyme production strains. A high level of synteny was observed with other aspergilli sequenced. Strong function predictions were made for 6,506 of the 14,165 open reading frames identified. A detailed description of the components of the protein secretion pathway was made and striking differences in the hydrolytic enzyme spectra of aspergilli were observed. A reconstructed metabolic network comprising 1,069 unique reactions illustrates the versatile metabolism of A. niger. Noteworthy is the large number of major facilitator superfamily transporters and fungal zinc binuclear cluster transcription factors, and the presence of putative gene clusters for fumonisin and ochratoxin A synthesis.

Fumonisin B2 production by Aspergillus niger.

Abstract: The carcinogenic mycotoxin fumonisin B2 was detected for the first time in the industrially important Aspergillus niger. Fumonisin B2, known from Fusarium verticillioides and other Fusaria, was detected in cultures of three full genome sequenced strains of A. niger, in the ex type culture and in a culture of F. verticillioides by electrospray LC-MS analysis of methanolic extracts from agar plugs of cultures grown on several substrates. Whereas F. verticillioides produced fumonisins B1, B2, and B3 on agar media based on plant extracts, such as barley malt, oat, rice, potatoes, and carrots, A. niger produced fumonisin B2 best on agar media with a low water activity, including Czapek yeast autolysate agar with 5% NaCl. Of the media tested, only rice corn steep agar supported fumonisin production by both F. verticillioides and A. niger. However, A. niger had a different regulation of fumonisin production and a different quantitative profile of fumonisins, producing only B2 as compared to F. verticillioides. Fumonisin production by A. niger, which is a widely occurring species and an extremely important industrial organism, will have very important implications for biotechnology and especially food safety. A. niger is used for the production of citric acid and as producer of extracellular enzymes, and also as a transformation host for the expression of heterologous proteins. Certain strains of A. niger produce both ochratoxin A and fumonisins, so some foods and feeds may potentially contain two types of carcinogenic mycotoxins from this species.

Antifungal activities of thyme, clove and oregano essential oils

Abstract: The antifungal potential of essential oils of oregano (Origanum vulgare), thyme (Thymus vulgaris) and clove (Syzygium aromaticum) was determined. To establish this antifungal potential, two molds related to food spoilage, Aspergillus niger and Aspergillus flavus, were selected. The agar dilution method was employed for the determination of antifungal activities. The three essential oils analyzed presented inhibitory effects on both molds tested. Oregano essential oil showed the highest inhibition of mold growth, followed by clove and thyme. Aspergillus flavus was more sensitive to thyme essential oil than A. niger. Clove essential oil was a stronger inhibitor against A. niger than against A. flavus.

An Overview of the Genus Aspergillus

Abstract: CONTENTS 1.1 Early History and Taxonomy 3 1.2 Industry 61.2.1 Koji 6 1.2.2 Citric Acid and Other Aspergillus niger Products 7 1.2.3 Aspergillus Secondary Metabolites 81.2.3.1 Lovastatin 8 1.2.3.2 Afl atoxin 81.3 Aspergillus as an Animal Pathogen 9 1.4 Genetics and Aspergillus 10 1.5 Genomics and the Future 11 References 11Few fungi are as important as members of the genus Aspergillus. This taxonomic group encompasses organisms whose characteristics are of high pathological, agricultural, industrial, pharmaceutical, scientifi c, and cultural importance. Superb agents of biodeterioration, aspergilli have been isolated from sources as varied as alligator nesting material, aviation fuel, Egyptian mummies, electrical fuses, plastic products, and old sauna boards. Indeed, this large and cosmopolitan group of molds is a major player in the ecosystem, involved in the degradation of a wide range of natural organic substrates, particularly plant materials. Aspergillus species are generalists in that they will grow and reproduce on many different carbon sources; they have an amazing nutritional fl exibility. The diversity of enzymes and organic acids used in nutrition is complemented by the metabolic capacity to secrete numerous low molecular weight secondary metabolites believed to be important in ecological signaling. Because these molds can be found almost everywhere on the planet, degrading both natural and human-made substrates, Aspergillus and human history have been intertwined intimately for centuries.

Aspergillus brasiliensis sp. nov., a biseriate black Aspergillus species with world-wide distribution

Abstract: A novel species, Aspergillus brasiliensis sp. nov., is described within Aspergillus section Nigri. This species can be distinguished from other black aspergilli based on intergenic transcribed region, β-tubulin and calmodulin gene sequences, by amplified fragment length polymorphism analysis and by extrolite profiles. A. brasiliensis isolates produced naphtho-γ-pyrones, tensidol A and B and pyrophen in common with Aspergillus niger and Aspergillus tubingensis, but also several unique compounds, justifying their treatment as representing a separate species. None of the isolates were found to produce ochratoxin A, kotanins, funalenone or pyranonigrins. The novel species was most closely related to A. niger, and was isolated from soil from Brazil, Australia, USA and The Netherlands, and from grape berries from Portugal. The type strain of Aspergillus brasiliensis sp. nov. is CBS 101740T (=IMI 381727T=IBT 21946T).

Control of Aspergillus niger with garlic, onion and leek extracts

Abstract: Antifungal activity of “Allium” vegetables that is garlic (Allium sativum L.), onion (Allium cepa L.) and leek (Allium porrum L.) were investigated against Aspergillus niger. Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) of aqueous, ethyl alcohol and acetone extracts were determined by disc diffusion and broth dilution methods in the test tubes. Onion extract with ethyl alcohol (275 mg/mL MFC), aqueous garlic extract (325 mg/mL MFC) and aqueous leek extract (900 mg/mL MFC) found the most inhibitory against A. niger.

Differential interaction of Aspergillus niger and Peniophora lycii phytases with soil particles affects the hydrolysis of inositol phosphates

Abstract: The effect of the soil environment on the mobility, stability and catalytic activity of phytase from two sources was compared, as these factors have important implications for the efficacy of enzyme function in soil. Phytase from an ascomycete fungus (Aspergillus niger) and a basidiomycete fungus (Peniophora lycii) was added to soil suspensions from three contrasting soils and activities in the solution and solid phase were monitored. The two enzymes were compared because the P. lycii phytase was known to have greater specific activity and a more acidic isoelectric point (pI) than A. niger and therefore predicted to have different adsorption characteristics. When added to soil suspensions buffered at pH 7.5, both phytases remained in solution in all of the soils. In contrast at near natural soil pH (pH 5.5), only the P. lycii phytase remained in solution, while the A. niger phytase was rapidly adsorbed to the soil solid phase. The extent of this adsorption was reduced, however, in a soil-dependent manner by prior addition of bovine serum albumin (BSA) to the soil suspensions. At the natural pH of the soil, the stability of the P. lycii phytase in soil solution was improved under sterile conditions, whereas degradation of the A. niger phytase was unaffected. Subsequently, P. lycii phytase was shown to be more effective at hydrolysing myo-inositol hexakisphosphate added to the soil. Moreover, the P. lycii phytase also hydrolysed more organic phosphate that was endogenous to a range of soils. This research indicates that the physicochemical properties of fungal phytases affect their mobility and temporal stability and their capacity to hydrolyse inositol phosphates in soil environments.

Solubilization of inorganic phosphates and plant growth promotion by Aspergillus niger

Abstract: Two of 187 fungal isolates (Aspergillus niger 1B and 6A) displaying superior phosphate (P) solubilization and hydrolytic enzyme secretion were studied using P forms of calcium (Ca-P), iron (Fe-P), and aluminum (Al-P). Phosphate solubilization in a sucrose-basal salt (SB) broth was increased and pH decreased by both isolates. In Ca-P medium, solubilization for 6A was approximately 322 μg P mL−1 and pH decreased by 4.2 units to 2.3 in 72 h. However, when pH value of the SB broth was lowered to 2.5 using HCl, 65.3 ± 0.4 μg mL−1 of P was released from Ca-P, whereas trace amounts of P were released from Fe-P and Al-P. Both isolates displayed enhanced Al-P solubilization using NH4Cl rather than KNO3 as the N source; final pH values were not significantly different. With Ca-P, gluconic acid was predominantly produced by 1B and 6A, whereas oxalic acid predominated with Fe-P and Al-P. Addition of gluconic acid (final concentration of 8.5 μmol mL−1) to Ca-P-supplemented SB lowered pH (2.9) and solubilized phosphate (146.0 ± 1.0 μg mL−1). Similarly, addition of oxalic acid (final concentration 6.6 μmol mL−1) to Ca-P- and Fe-P-amended media solubilized P (60.2 ± 0.9 and 21.6 ± 2.1 μg mL−1, respectively), although these quantities were significantly lower than those detected in unamended SB. The presence of unidentified P solubilized compound(s) in the dialyzed (MW>500) supernatant warrants further study. In pot experiments, significant increases in plant (Brassica chinensis Linn.) dry weight and N and P contents were observed with the addition of isolate 6A, when a small amount of organic fertilizer together with either rock phosphate (South African apatite) or Ca-P served as the main P sources.

Cellulolytic enzymes on lignocellulosic substrates in solid state fermentation by Aspergillus niger.

Abstract: The production of cellulolytic enzymes by Aspergillus niger on lignocellulosic substrates groundnut fodder, wheat bran, rice bran and sawdust in solid state fermentation in a laboratory scale was compared. Czapek Dox liquid broth amended with cellulose (0.5%) was used to moisten lignocellulosic solid supports for cultivation of Aspergillus niger. The production of filter paperase, carboxymethyl cellulase and -glucosidase were monitored at daily intervals for 5 days. The peak production of the enzymes occurred within 3 days of incubation. Among solid supports used in the study, wheat bran was the best solid matrix followed by groundnut fodder in production of cellulolytic enzymes in solid state fermentation. Groundnut fodder supported significant production of FPase (2.09 FPU/g), CMCase (1.36 U/g) and -glucosidase activity (0.0117 U/g) in solid state fermentation. Considerable secretion of protein (5.10 mg/g) on groundnut fodder at peak time interval 1st day of incubation was recorded.

The antifungal protein AFP from Aspergillus giganteus inhibits chitin synthesis in sensitive fungi.

Abstract: The antifungal protein AFP from Aspergillus giganteus is highly effective in restricting the growth of major human- and plant-pathogenic filamentous fungi. However, a fundamental prerequisite for the use of AFP as an antifungal drug is a complete understanding of its mode of action. In this study, we performed several analyses focusing on the assumption that the chitin biosynthesis of sensitive fungi is targeted by AFP. Here we show that the N-terminal domain of AFP (amino acids 1 to 33) is sufficient for efficient binding of AFP to chitin but is not adequate for inhibition of the growth of sensitive fungi. AFP susceptibility tests and SYTOX Green uptake experiments with class III and class V chitin synthase mutants of Fusarium oxysporum and Aspergillus oryzae showed that deletions made the fungi less sensitive to AFP and its membrane permeabilization effect. In situ chitin synthase activity assays revealed that chitin synthesis is specifically inhibited by AFP in sensitive fungi, indicating that AFP causes cell wall stress and disturbs cell integrity. Further evidence that there was AFP-induced cell wall stress was obtained by using an Aspergillus niger reporter strain in which the cell wall integrity pathway was strongly induced by AFP.

Xylanase production by Aspergillus niger ANL 301 using agro - wastes

Abstract: Xylanase production by wild-type Aspergillus niger ANL301, newly isolated from wood-waste, was monitored at 24 h intervals for a period 168 h in media containing different carbon sources. The carbon sources were oat-spelt xylan (Fluka) and three agro-wastes (sawdust, sugarcane pulp and wheat bran). Highest xylanase activity of 6.47 units/mL was obtained at 96 h in media containing wheat bran as sole carbon source. Maximum activity value for the media containing sugarcane pulp was 0.95 units/mL obtained also at 96 h. Sawdust and oat spelt xylan gave the peak enzyme activities of 0.65 and 0.80 units/mL respectively at 120 h. High protein yield was obtained in media containing the agro-wastes, with wheat bran giving the highest value of 1.14 mg/mL at 96 h. The maximum specific xylanase activities were 3.86, 3.37, 5.69, and 9.36 units/ mg protein for sawdust, sugarcane pulp, wheat bran and oat spelt xylan, respectively. Out of the three agro-wastes used in this study, wheat bran holds greatest promise for low cost production of the xylanase enzyme.

Production of fructose from inulin using mixed inulinases from Aspergillus niger and Candida guilliermondii

Abstract: Two new effective microbial producers of inulinases were isolated from Jerusalem artichoke tubers grown in Thailand and identified as Aspergillus niger TISTR 3570 and Candida guilliermondii TISTR 5844. The inulinases produced by both these microor- ganisms were appropriate for hydrolysing inulin to fructose as the principal product. An initial inulin concentration of ~100 g l -1 and the enzyme concentra- tion of 0.2 U g -1 of substrate, yielded 37.5 g l -1 of fructose in 20 h at 40� C when A. niger TISTR 3570 inulinase was the biocatalyst. The yield of fructose on inulin was 0.39 g g -1 . Under identical conditions, the yeast inulinase afforded 35.3 g l -1 of fructose in 25 h. The fructose yield was 0.35 g g -1 of substrate. The fructose productivities were 1.9 g l -1 h -1 and 1.4 g l -1 h - 1 for the mold and yeast enzymes, respectively. After 20 h of reaction, the mold enzyme hydrolysate con- tained 53% fructose and more than 41% of initial inulin had been hydrolysed. Using the yeast enzymes, the hydrolysate contained nearly 38% fructose at 25 h and nearly 36% of initial inulin had been hydrolysed. The A. niger TISTR 3570 inulinases exhibited both endo- inulinase and exo-inulinase activities. In contrast, the yeast inulinases displayed mainly exo-inulinase activity. The mold and yeast crude inulinases mixed in the activity ratio of 5:1 proved superior to individual crude inulinases in hydrolysing inulin to fructose. The enzyme mixture provided a better combination of endo- and exo-inulinase activities than did the crude extracts of either the mold or the yeast individually.

Structure−Antifungal Activity Relationship of Cinnamic Acid Derivatives

Abstract: A structure-antifungal activity relationship (SAR) study of 22 related cinnamic acid derivatives was carried out. Attention was focused on the antifungal activities exhibited against Aspergillus flavus, Aspergillus terreus, and Aspergillus niger. (E)-3-(4-methoxy-3-(3-methylbut-2-enyl)phenyl)acrylic acid (16) exhibited antifungal activity against A. niger, comparable to that of miconazole and a significant antifungal effect against A. flavus and A. terreus as well. A structure-activity relationship (SAR) study of related cinnamic acid derivatives has allowed a model to be proposed for the recognition of the minimal structural requirements for the antifungal effect in this series.

Isolation and purification of an enzyme hydrolyzing ochratoxin A from Aspergillus niger

Abstract: Ochratoxin A is a mycotoxin produced by several Aspergillus and some Penicillium species which may be present in food and feed products. It can be enzymatically hydrolyzed into ochratoxin α and l-β-phenylalanine, thereby decreasing its toxicity. The ochratoxin A degradation capacity of Aspergillus niger is well known and here we report the isolation and purification of a novel enzyme from A. niger that hydrolyzes this mycotoxin. A wheat germ medium supplemented with ochratoxin A was used to produce the enzyme, which was purified from culture filtrate by acetone precipitation and anion exchange chromatography. An overall purification of 2.5-fold with a recovery of 68% and a final specific activity of 36 U/mg was obtained. The enzyme is a metalloenzyme as it was inhibited at 10 mM EDTA, whereas PMSF had no effect. The ochratoxin A hydrolytic enzyme presented a Vmax of 0.44 μM/min and a Km of 0.5 mM when the reaction was carried out at pH 7.5 and 37°C.

Adopting Selected Hydrogen Bonding and Ionic Interactions from Aspergillus fumigatus Phytase Structure Improves the Thermostability of Aspergillus niger PhyA Phytase

Abstract: Although it has been widely used as a feed supplement to reduce manure phosphorus pollution of swine and poultry, Aspergillus niger PhyA phytase is unable to withstand heat inactivation during feed pelleting. Crystal structure comparisons with its close homolog, the thermostable Aspergillus fumigatus phytase (Afp), suggest associations of thermostability with several key residues (E35, S42, R168, and R248) that form a hydrogen bond network in the E35-to-S42 region and ionic interactions between R168 and D161 and between R248 and D244. In this study, loss-of-function mutations (E35A, R168A, and R248A) were introduced singularly or in combination into seven mutants of Afp. All seven mutants displayed decreases in thermostability, with the highest loss (25% [P < 0.05]) in the triple mutant (E35A R168A R248A). Subsequently, a set of corresponding substitutions were introduced into nine mutants of PhyA to strengthen the hydrogen bonding and ionic interactions. While four mutants showed improved thermostability, the best response came from the quadruple mutant (A58E P65S Q191R T271R), which retained 20% greater (P < 0.05) activity after being heated at 80°C for 10 min and had a 7°C higher melting temperature than that of wild-type PhyA. This study demonstrates the functional importance of the hydrogen bond network and ionic interaction in supporting the high thermostability of Afp and the feasibility of adopting these structural units to improve the thermostability of a homologous PhyA phytase.

Mixed substrate solid state fermentation for production and extraction of lipase from Aspergillus niger MTCC 2594.

Abstract: A novel mixed substrate solid-state fermentation (SSF) process has been developed for Aspergillus niger MTCC 2594 using wheat bran (WB) and gingelly oil cake (GOC) and the results showed that addition of GOC to WB (WB : GOC, 3 : 1, w/w) increased the lipase activity by 36.0% and the activity was 384.3+/-4.5 U/g dry substrate at 30 degrees C and 72 h. Scale up of lipase production to 100 g and 1 kg tray-level batch fermentation resulted in 95.0% and 84.0% of enzyme activities respectively at 72 h. A three-stage multiple contact counter-current extraction yielded 97% enzyme recovery with a contact time of 60 min. However, extraction by simple percolation and plug-flow methods resulted in decreased enzyme recoveries. The mixed substrate SSF process has resulted in a significant increase in specific activity (58.9%) when compared to a submerged fermentation (SmF) system. Furthermore, an efficient process of extraction has been standardized with this process. Use of GOC along with WB as potential raw materials for enzyme production could be of great commercial significance. This is the first report on the production and extraction of lipase from Aspergillus niger using mixed solid substrates, WB and GOC, which are potential raw materials for the production of enzymes and other value-added products.

Antimicrobial activity of methanol extract of Origanum majorana L. (Sweet marjoram).

Abstract: In-vitro microbicidal activity of the methanol extract of Origanum majorana L was tested against seven fungi (Fusarium solani, Candida albicans, Aspergillus niger, A parasiticus, Rhizopus oryzae, Rhizoctonia otyzae-sativae and Altemaria brassicicola) and six bacteria (Bacillus subtilis, B megaterium, Escherichia coil, Proteus vulgaris, Pseudomonas aeruginosa and Staphylococcus aureus) The methanol extract of O majorana can be used as an effective herbal protectant against different pathogenic bacteria and fungi High toxicity against the growth of Aspergillus niger was diagnosed

Directed evolution of glucose oxidase from Aspergillus niger for ferrocenemethanol-mediated electron transfer.

Abstract: A directed evolution protocol was developed for glucose oxidase (GOx) from Aspergillus niger that mimics applications conditions and employs a well-known mediator, oxidized ferrocenemethanol, in a medium throughput screen (96-well plate format). Upon reduction, oxidized ferrocenemethanol shows a color change from blue to pale yellow that can be recorded at 625 nm. Under optimized screening conditions, a CV of less than 20% was achieved in 96-well microtiter plates. For validating the screening system, two mutant libraries of GOx were generated by standard error-prone PCR conditions (0.04 mM MnCl(2)) and Saccharomyces cerevisiae was employed as host for secreted GOx expression. Two screening of approximately 2000 GOx mutants yielded a double mutant (T30S I94V) with improved pH and thermal resistance. Thermal resistance at a residual activity of 50% was increased from 58 degrees C (wild type, WT) to 62 degrees C (T30S I94V) and pH stability was improved at basic pH (pH 8-11). K(m) for glucose remained nearly unchanged (20.8 mM WT; 21.3 mM T30S I94V) and k(cat) increased (69.5/s WT; 137.7/s T30S I94V).