Effect of Aspergillus niger prolyl endopeptidase in patients with celiac disease on a long-term gluten-free diet

BACKGROUND The gluten-free diet(GFD)has limitations,and there is intense research in the development of adjuvant therapies.AIM To examine the effects of orally administered Aspergillus niger prolyl endopeptidase protease(AN-PEP)on inadvertent gluten exposure and symptom prevention in adult celiac disease(CeD)patients following their usual GFD.METHODS This was an exploratory,double-blind,randomized,placebo-controlled trial that enrolled CeD patients on a long-term GFD.After a 4-wk run-in period,patients were randomized to 4 wk of two AN-PEP capsules(GliadinX;AVI Research,LLC,United States)at each of three meals per day or placebo.Outcome endpoints were:(1)Average weekly stool gluten immunogenic peptides(GIP)between the run-in and end of treatments and between AN-PEP and placebo;(2)celiac symptom index(CSI);(3)CeD-specific serology;and(4)quality of life.Stool samples were collected for GIP testing by ELISA every Tuesday and Friday during run-ins and treatments.RESULTS Forty patients were randomized for the intention-to-treat analysis,and three were excluded from the per-protocol assessment.Overall,628/640(98.1%)stool samples were collected.GIP was undetectable(<0.08μg/g)in 65.6%of samples,and no differences between treatment arms were detected.Only 0.5%of samples had GIP concentrations sufficiently high(>0.32μg/g)to potentially cause mucosal damage.Median GIP concentration in the AN-PEP arm was 44.7%lower than in the run-in period.One-third of patients exhibiting GIP>0.08μg/g during run-in had lower or undetectable GIP after AN-PEP treatment.Compared with the run-in period,the proportion of symptomatic patients(CSI>38)in the AN-PEP arm was significantly lower(P<0.03).AN-PEP did not result in changes in specific serologies.CONCLUSION This exploratory study conducted in a real-life setting revealed high adherence to the GFD.The AN-PEP treatment did not significantly reduce the overall GIP stool concentration.However,given the observation of a significantly lower prevalence of patients with severe symptoms in the AN-PEP arm,further clinical research is warranted.

Secondary pulmonary infection by Fusarium solani and Aspergillus niger during systemic steroid treatment for COVID-19:A case report

BACKGROUND Coronavirus disease 2019(COVID-19)-associated invasive pulmonary aspergillosis presents a diagnostic challenge due to its non-specific clinical/imaging features,as well as the fact that the proposed clinically diagnostic algorithms do not necessarily apply to COVID-19 patients.In addition,Fusarium spp.is a rare cause of opportunistic life-threatening fungal infections.Disseminated Fusarium infection in an immunocompromised host is intractable,with a high likelihood of resulting mortality.To our knowledge,this is the first case of secondary pulmonary infection by Fusarium solani(F.solani)and Aspergillus niger(A.niger)during systemic steroid treatment for COVID-19.CASE SUMMARY A 62-year-old male was transported to our hospital by ambulance with a complaint of fever and dyspnea.We established a diagnosis of pneumococcal pneumonia,complicated with COVID-19 and septic shock,together with acute renal failure.He was admitted to the intensive care unit,to be treated with piperacillin/tazobactam,vancomycin,and 6.6 mg per day of dexamethasone sodium phosphate,along with noradrenaline as a vasopressor,ventilator management,and continuous hemodiafiltration.His condition improved,and we finished the vasopressor on the fifth hospital day.We administered dexamethasone for ten days,and finished the course of treatment.On the eleventh day,patient respiratory deterioration was observed,and a computed tomography scan showed an exacerbation of bilateral ground-glass-opacity-like consolidation,together with newly appeared cavitary lesions in the lung.we changed antibiotics to meropenem plus vancomycin.In addition,a fungal infection was considered as a possibility based on microscopic findings of sputum,and we began coadministration of voriconazole.However,the pneumonia worsened,and the patient died on the seventeenth day of illness.Later,F.solani and A.niger were identified from sputum collected on the twelfth day.It was believed that he developed a cell-mediated immune deficiency during COVID-19 treatment,which led to the complication of pneumonia caused by the above-mentioned fungi,contributing to his death.CONCLUSION Because early initiation of intense antifungal therapy offers the best chance for survival in pulmonary fusariosis,computed tomography scans and appropriate microbiologic investigations should be obtained for severely immunocompromised patients.

Kinetics of Bioremediation of Oil Contaminated Water Dispersed by Environment-Friendly Bacteria (Pseudomonas aeruginosa) and Fungi (Aspergillus niger)

The comparative effectiveness of remediating water polluted with crude oil, using environment-friendly bacteria (Pseudomonas aeruginosa) and fungi (Aspergillus niger) were investigated. The samples were separately treated with Aspergillus niger and Pseudomonas aeruginosa. The bioremediation kinetic efficiency for these systems was studied. At the end of the bioremediation periods, the oil and grease content of the samples decreased from 47.0 mg/L in the untreated sample to 7.0 mg/L after 20 days when inoculated with bacteria while the sample inoculated with fungi decreased to 10.0 mg/L. Post analysis when inoculated with bacteria showed a fall in the value of the biological oxygen demand (BOD) from 73.84 mg/L to 33.28 mg/L after 20 days, while, the fungi inoculated sample showed a reduction from 73.84 mg/L to 38.48 mg/L. The biodegradation process with the bacteria was consistent with the pseudo-first-order model with a rate constant of 0.0891 day-1, while the biodegradation process with the fungi was consistent with the first order reaction model with a rate constant of 0.422 day-1. The degree of degradation after the 20th day of inoculation with Pseudomonas aeruginosa was 85.11%, while with Aspergillus niger was 78.72%. Thus, the results obtained showed that, Pseudomonas aeruginosa performed better than Aspergillus niger. The bioremediation data with fungi fitted the first-order model, while that of the bacteria fitted the pseudo-first-order model. Therefore, the data obtained in this study could be applied in the design of a bioremediation system for potential application to remediation of crude oil polluted water.

Solid state fermentation of rapeseed cake with Aspergillus niger for degrading glucosinolates and upgrading nutritional value

Background： Rapeseed cake is a good source of protein for animal feed but its utilization is limited due to the presence of anti-nutritional substances, such as glucosinolates （GIs）, phytic acid, tannins etc. In the present study, a solid state fermentation （SSF） using Aspergillus niger was carried out with the purpose of degrading glucosinolates and improving the nutritional quality of rapeseed cake （RSC）. The effects of medium composition and incubation conditions on the GIs content in fermented rapeseed cake （FRSC） were investigated, and chemical composition and amino acid in vitro digestibility of RSC substrate fermented under optimal conditions were determined. Results： After 72 h of incubation at 34℃, a 76.89% decrease in GIs of RSC was obtained in solid medium containing 70% RSC, 30% wheat bran at optimal moisture content 60% （w/w）. Compared to unfermented RSC, trichloroacetic acid soluble protein （TCA-SP）, crude protein and ether extract contents of the FRSC were increased （P〈 0.05） 103.71, 23.02 and 23.54%, respectively. As expected, the contents of NDF and phytic acid declined （P〈 0.05） by 9.12 and 44.60%, respectively. Total amino acids （TAA） and essential amino acids （EAA） contents as well as AA in vitro digestibility of FRSC were improved significantly （P 〈 0.05）. Moreover, the enzyme activity of endoglucanase, xylanase, acid protease and phytase were increased （P 〈 0.05） during SSF. Conclusions： Our results indicate that the solid state fermentation offers an effective approach to improving the quality of proteins sources such as rapeseed cake.

A new naphthoquinoneimine derivative from the marine algal-derived endophytic fungus Aspergillus niger EN-13

Cultivation of an endophytic fungus Aspergillus niger EN-13 that was isolated from the inner tissue of the marine brown alga Colpomenia sinuosa resulted in the characterization of a new naphthoquinoneimine derivative, namely, 5,7-dihydroxy-2-[1-（4- methoxy-6-oxo-6H-pyran-2-yl）-2-phenylethylamino]-[ 1,4]naphthoquinone. The structure of the new compound was established on the basis of various NMR spectroscopic analyses including 2D NMR techniques, El-MS, and HR-ESI-MS. This compound displayed moderate antifungal activity.

Salt-tolerant mechanism of marine Aspergillus niger cellulase cocktail and improvement of its activity

The cellulase cocktail produced by marine Aspergillus niger exhibits a property of salt-tolerance,which is of great potential in cellulose degradation in high salt environment.In order to explain the mechanism on the salttolerance of the cellulase cocktail produced by marine A.niger,six cellulase components(AnCel6,AnCel7A,AnCel7B,AnEGL,AnBGL1 and AnBGL2)were obtained by directed expression.Studies on their enzymatic properties revealed that oneβ-glucosidase(AnBGL2)and one endoglucanase(AnEGL)exhibited an outstanding salttolerant property,and one cellobiohydrolase(AnCel7B)exhibited a certain salt-tolerant property.Subsequent study revealed that the salt-tolerant An EGL and AnCel7B endowed the cellulase cocktail with stronger salttolerant property,while the salt-tolerant An BGL2 had no positive effect.Moreover,after overexpression of AnCel6,AnCel7A,AnCel7B and AnEGL,the activity of cellulase cocktail increased by 80%,70%,63%and 68%,respectively.However,the activity of cellulase cocktail was not improved after overexpression of AnBGL1 and AnBGL2.After mixed-strain fermentation with cellobiohydrolase recombinants(cel6 a,cel7a and cel7b recombinants)and endoglucanase recombinant(egl recombinant),the the activity of cellulase cocktail increased by 114%,102%and91%,respectively.

Optimization of Fermentation Conditions for Xylanase Production by Aspergillus niger NS-1

In this study, a xylanase-produeing Aspergillus niger strain, NS-1, was screened and isolated from agricultural and forestry wastes. Based on single-fac- tor experiments, the effects of different carbon sources, composite carbon sources, nitrogen sources, calcium carbonate concentrations, initial pH and surfactants on xylanase production by A. niger NS-1 were investigated. The results indicated that the most appropriate carbon source was corncobs ; the best composite carbon source was corncobs ＋ xylan, which was conducive to xylanase secretion; the most suitable nitrogen source was ammonium sulfate. Xylanase activity reached the highest in the medium added with 1.5% calcium carbonate and SDS as a surfactant with an initial pH of 5.0. This study provided the basis for the production of high-activity xylanase.

The Site-Specific Hydrolysis of l-β-Hydroxybaccatin Ⅱ by Aspergillus Niger

The site-specific microbiological hydrolysis of a natural 1β-hydroxybaccatin I, with the culture of Aspergillus niger. is described.

Microbial leaching of chromite overburden from Sukinda mines,Orissa,India using Aspergillus niger

Leaching of nickel and cobalt from two physical grades （S1, 125-190 μm, coarser and S3, 53-75 μm, finer） of chromite overburden was achieved by treating the overburden （2% pulp density） with 21-d culture filtrate of an Aspergillus niger strain grown in sucrose medium. Metal dissolution increases with ore roasting at 600℃ and decreasing particle size due to the alteration of microstructural properties involving the conversion of goethite to hematite and the increase in surface area and porosity as evident from X-ray diffraction （XRD）, thermogravimetry-differential thermal analysis （DT- TGA）, and field emission scanning electron microscopy （FESEM）. About 65% Ni and 59% Co were recovered from the roasted S3 ore employing bioleaching against 26.87% Ni and 31.3% Co using an equivalent amount of synthetic oxalic acid under identical conditions. The results suggest that other fungal metabolites in the culture filtrate played a positive role in the bioleaching process, making it an efficient green approach in Ni and Co recovery from lateritic chromite overburden.

Removal process and mechanism of hexavalent chromium by adsorption-coupled reduction with marine-derived Aspergillus niger mycelial pellets

In order to remove hexavalent chromium(Cr(Ⅵ))from solutions efficiently,the mycelial pellets with a marine-derived fungus Aspergillus niger as a biosorbent were prepared.The effects of removal process parameters such as solution pH,initial Cr(Ⅵ)concentration and biomass concentration on Cr(Ⅵ)removal process were investigated.The results showed that Cr(Ⅵ)removal rate up to 100%could be achieved under optimized conditions,which indicated the excellent Cr(Ⅵ)removal performance of the Aspergillus niger pellets.As a more important point,the Cr(Ⅵ)removal mechanism was studied,and the results revealed that Cr(Ⅵ)removal was achieved in the adsorption-coupled reduction process.A little of Cr(Ⅵ)was reduced to less toxic trivalent chromium(Cr(Ⅲ))in solution,while some was absorbed on the surface of mycelial pellets.Then they may be reduced on the surface or transferred into cells and then be reduced.The marine-derived A.niger mycelial pellets show properties of easy preparation and separation and cost effectiveness,which are potential biosorbent and reductant in the treatment of trace chromate containing wastewater.

Pumpkin-like MoP-MoS_(2)@Aspergillus niger spore-derived N-doped carbon heterostructure for enhanced potassium storage

Biomass-derived carbon materials are widely applied in the energy storage and conversion fields due to their rich sources,low price and environmental friendliness.Herein,a unique pumpkin-like MoPMoS_(2)@Aspergillus niger spore-derived N-doped carbon(SNC)composite has been prepared via a simple hydrothermal and subsequent phosphorization process.Interestingly,the resulting MoP-MoS_(2)@SNC well inherits the pristine morphology of spore carbon,similar to the natural pumpkin,with hollow interiors and uneven protrusions on the surface.The special structure allows it to have sufficient space to fully contact the electrolyte and greatly reduces the ion transport distance.The theory calculations further demonstrate that the formed MoP-MoS_(2)heterostructure can enhance the adsorption of K ions and electronic couplings.With these unique advantages,the MoP-MoS_(2)@SNC anode for potassium storage shows a high reversible capability of 286.2 mAh g&(-1) at 100 mA g^(-1) after 100 cycles and superior rate performance.The enhanced electrochemical performance is mainly related to the unique pumpkin-like morphology of SNC and the construction of MoP-MoS_(2)heterostructure,as well as their perfect coupling.This study provides a feasible design idea for developing green,low-cost,and high-performance electrode materials for next-generation energy storage.

Effect of Aspergillus niger NBC001 on the soybean rhizosphere microbial community in a soybean cyst nematode-infested field

Soybean cyst nematode(SCN,Heterodera glycines Ichinohe)is one of the most important pests causing considerable damage to soybean(Glycine max(L.)Merr.)around the world.Biocontrol provides a strategy for sustainable nematode control.Previously,Aspergillus niger NBC001 was isolated from the cysts Heterodera spp.and able to control H.glycines and promote the growth of soybean in a pot experiment.In this study,the effects of NBC001 on H.glycines density and on the soybean rhizosphere microbial community in a soybean cyst nematode-infested field were studied.The results showed that NBC001 could suppress H.glycines by 31.7%in the field.High-throughput sequencing analysis showed that NBC001 had no significant influence on soybean rhizosphere microbial community structure,indicating that seed coatdressing with the concentrated culture filtrate of NBC001 was safe for the soil ecological environment.In addition,high-throughput sequencing results demonstrated that at 10 days post transplantation,NBC001 increased the abundances of Actinobacteria and Acidobacteria,but decreased the abundances of Bacteroidetes and Gemmatimonadetes at the phylum level.Meanwhile,the abundances of Phyllobacterium,Ralstonia and H16 were increased,while the abundances of Adhaeribacter,Gemmatimonas,Sphingomonas,Flavisolibacter\Nere suppressed by application of NBC001.However,at 90 days post transplantation,NBC001 only increased the abundances oi Aeromicrobium and RB41 whereas it decreased the abundance of H16.The results indicated that application of NBC001 increased the relative abundances of the beneficial microorganisms such as Actinobacteria,Acidobacteria,Aeromicrobium and Phyllobacterium in the soil.In summary,NBC001 is an eco-friendly biocontrol agent for H.glycines control.

Chitosan Obtained from Cell Wall of Aspergillus Niger Mycelium

Chitin from cell walls of Aspergillus Niger mycelium was prepared. A new method for the preparation of high deacetylation degree chitosan was studied in a dilute sodium hydroxide solution at a high pressure. The experimental results indicate that the deacetylation degree of the chitosan can reach 80% under the condition of a 5.00 mol/L sodium hydroxide solution at 0.1 MPa of pressure for 1 h. This method shows the advantages of the applications in the industry production and environment protection.

Damaging Effect of Low Energy N^+ Implantation on Aspergillus niger Spores

The mutant effects of a keV range nitrogen ion （N＋） beam on enzyme-producing probiotics were studied, particularly with regard to the induction in the genome. The electron spin resonance （ESR） results showed that the signal of ESR spectrum existed in both implanted and non-implanted spores, and the yields of free radicals increased in a dose-dependent manner. The ionic etching and dilapidation of cell wall could be observed distinctly through the scanning electron microscope （SEM）. The mutagenic effect on genome indicated that N＋ implantation could make base mutation. This study provided an insight into the roles low-energy ions might play in inducing mutagenesis of micro-organisms.

Effect of Different Cultural Conditions for Phytase Production by Aspergillus niger in Submerged Fermentation

In agriculture, phytase is one of the most important monogastric animal sources of nutrient components because it effectively catalyzes the release of phosphate from phytate and phosphorylated compounds. In present work, Aspergillus niger strain (own collection) was used. Various physical and chemical factors have been known to affect the growth and the production of phytase. The effect of carbon and nitrogen sources, temperature and pH for extra cellular phytase production was investigated. Maximal phytase activity of Aspergillus niger was detected in media with 1.0% sucrose as a carbon source. Among the inorganic and organic nitrogen sources, ammonium nitrate in concentration of 0.5% was found to be a favorable nitrogen source for phytase production in Aspergillus niger. Optimum temperature and pH for phytase production by Aspergillus niger were 30°C and 5.5.

Fungitoxicity of Trichoderma longibrachiatum (Rifai) Metabolites against Fusarium oxysporum, Aspergillus niger and Aspergillus tamarii

An experiment was carried out to evaluate the fungitoxic effect of Trichoderma longibrachiatum (Rifai) metabolite on F. oxysporum, A. niger and A. tamarii. The fungi were collected from the International Institute of Tropical Agriculture (IITA) and Nigerian Institute of Science Laboratory Technology (NISLT). T. longibrachiatum was cultured on 1/4 strength Potato Dextrose Broth (PDB) following standard procedures. Its metabolite was extracted using 50 ml n-hexane with 50 ml Potato Dextrose Broth (PDB). The metabolite was purified by filter sterilization using a sterile 0.22 millipore filter disc after centrifuging at 900 rpm for 20 minutes. Petri plates of each fungus were later impregnated in triplicates with the T. longibrachiatum metabolites using four concentrations (10%, 25%, 50% and 100%), and three volumes (1 ml, 2 ml, and 3 ml). Petri plates of fungi without the metabolite and Petri Plates of fungi with n-hexane and PDB served as control. All Petri Plates were incubated at 28°C for 7 days. Radial and diametric growth of each fungus on all Petri Plates were taken daily at 24 hours intervals. Data obtained were analysed using SAS (version 9.3). Growth inhibition of F. oxysporum, A. niger and A. tamarii was significantly higher than in control in that order (P ≤ 0.05). Inhibition of the fungi by metabolites extracted with both PDB and n-hexane was significantly better than in control. Generally, inhibition by metabolite extracted with PDB was better than that extracted with n-hexane. Growth inhibition at all the concentrations of the metabolite was significantly better than in the control (P ≤ 0.05). Generally, inhibition at 100%, 50% and 25% concentrations was better than that at 10% concentration of the metabolite. Trichoderma longibrachiatum metabolite can thus be said to possess promising fungitoxic potential even at concentrations as low as 25%. Both PDB and n-hexane are good extraction media for the metabolite.

Study on Removal of Pb and Hg from MSWI Fly Ash by Aspergillus niger Leaching

Fly ash from a municipal solid waste incineration plant in Haikou was taken as the research object. The biological toxicity of its leachate was determined by luminescent bacteria method. The leaching toxicity of the leachate was determined by atomic fluorescence spectrometry. The results showed that the leaching with Aspergillus niger had good removal effect on As and Hg, and the leached fly ash and leachate could meet the safe discharge standard. 37 ℃ was the best temperature for A. niger leaching. The concentrations of As and Hg in fly ash leachate were reduced to 0.438 and 0.053 g/L, respectively after 5 d of leaching by A. niger at 37 ℃. The luminous intensity of leachate increased with the increase of the number of days. Therefore, the leaching with A. niger can effectively remove As and Hg in fly ash from municipal solid waste incineration, and the leachate can also meet the safe discharge standard.

Microbial transformation for Region-selective Hydroxylation of Z-Butylidenephthalide by Aspergillus niger CGMCC 3.739

Microbial transformation is a useful tool for structure modification of active natural products.Z-Butylidenephthalide is the main active constituent of Angelica sinensis.In the present work,a pair of new oxidized metabolites,namely(-)(11R)-(Z)-11-hydroxy-butylidenephthalide(1a)and(+)(11S)-(Z)-11-hydroxy-butylidenephthalide(1b),were obtained from microbial transformation for region-selective 11-hydroxylation of Z-butylidenephthalide by Aspergillus niger CGMCC 3.739.Their structures,including absolute configurations,were elucidated by extensive analysis of HR-ESI-MS,NMR spectra,and the modified Mosher’s method.This is the first study on the microbial transformation of Z-butylidenephthalide,and these findings offer a tool for region-selective 11-hydroxylation of Z-butylidenephthalide.

Screening of genes of secreting acetic acid from Aspergillus niger H1 to improve phosphate solubilization

A primary cDNA library of Aspergillus niger H1 was constructed using the switching mechanism at the 5′ end of the RNA transcript（SMART） technique. A total of 169 clones exhibited halos when grown on tricalcium phosphate medium, and the H-46 clone displayed a clear halo. The full-length c DNA of the clone H-46 clone was 1 407 bp in length with a complete open reading frame（ORF） of 816 bp, and it encoded a protein that contained 272 amino acids. Multiple alignment analysis revealed a high degree of homology between the ORFs of the H-46 clone and the Bax inhibitor family（BI-1-like） proteins of other fungi. Acetic acid was secreted by Escherichia coli DH5α that express the BI-1-like gene. The level attained was 492.52 mg L^（-1）, which was associated with the release of 0.212 mg m L^（-1） of soluble phosphate at 28 h. These results showed that the heterologous expression of BI-1-like genes in Eschericha coli DH5α increased the secretion of acetic acid by altering the membrane permeability and enhancing the solubility of phosphate（P）.