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.

Aspergillus niger prolyl endopeptidase in celiac disease

We comment here on the article by Stefanolo et al entitled“Effect of Aspergillus niger prolyl endopeptidase in patients with celiac disease on a long-term gluten-free diet”,published in the World Journal of Gastroenterology.Celiac disease is a well-recognized systemic autoimmune disorder.In genetically susceptible people,the most evident damage is located in the small intestine,and is caused and worsened by the ingestion of gluten.For that reason,celiac patients adopt a gluten-free diet(GFD),but it has some limitations,and it does not prevent re-exposure to gluten.Research aims to develop adjuvant therapies,and one of the most studied alternatives is supplementation with Aspergillus niger prolyl endopeptidase protease(AN-PEP),which is able to degrade gluten in the stomach,reducing its concentration in the small intestine.The study found a high adherence to the GFD,but did not address AN-PEP as a gluten immunogenic peptide reducer,as it was only tested in patients following a GFD and not in gluten-exposing conditions.This study opens up new research perspectives in this area and shows that further study is needed to clarify the points that are still in doubt.

Enzymatic Characterization of Pectinex XXL,a Pectinase Produced by Aspergillus niger,and Its Application in Fruit Juice Production

Pectinex XXL,a commercially prepared pectinase,was investigated for its potential application in the fruit juice industry.Polygalacturonic acid was used as the substrate for determining the enzymatic properties of Pectinex XXL using the DNS method.According to the results,the optimal pH for Pectinex XXL activity was 4.5,and the enzyme was stable in the pH range of 3.0~4.5.The optimal pH and pH stability range are consistent with those of some tropical and subtropical fruits.The optimal temperature for Pectinex XXL activity was 60℃,and the enzyme remained stable after one hour in a water bath set at 40℃.Additionally,the enzymatic activity was not inhibited in the presence of 1 mmol/L of Na^(+),Mg^(2+),Ba^(2+),Co^(2+),Zn^(2+),and Fe^(2+),whereas it was slightly inhibited in the presence of 2 mmol/L of K^(+)and Fe^(2+)and partially inhibited in the presence of 1 and 2 mmol/L of Ca^(2+)and Mn^(2+),demonstrating its good stability in acids and excellent thermal catalytic performance.Based on the above experimental results,depectinization experiments were performed on plantain and cherry tomato juices using different amounts of Pectinex XXL.After one hour reaction with 16 U/mL of the enzyme,the yields of the plantain and cherry tomato juices were substantially increased by 119.03%and 15.97%,respectively,while their light transmittance was remarkably enhanced by 37.65%and 12.35%,respectively.Furthermore,the enzyme reduced the viscosity of the plantain and cherry tomato juices by 88.29%and 29.50%,respectively.The juice production experiments confirmed that this enzyme can significantly improve the yield and light transmittance of plantain juice,while effectively reducing its viscosity.These findings indicate the potential of Pectinex XXL in the industrial production of plantain juice.

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.

Inducing and Screening of High Producing Pectinase Aspergillus niger Strain

[Objective]The aim was to induce and screen the high producing pectinase Aspergillus niger Strain based on the original preservation strains.[Method]The original strain was induced by ultraviolet,and the highst enzyme activity and cultivated time were detected through the inspection of transparent circle and enzyme activity determination of flask fermentation.[Result] The enzyme activity of strain D1-4 achieved its highest after cultivated for 96 h in suitable conditions,which was 141.13 U/ml.[Conclusion] The induced strain D1-4 had the strong ability of producing pectinase.

Improvement of Cellulase Producing Capacity of Aspergillus niger by Ultraviolet Mutation

[Objective] The research aimed to breed the high-yield production strain of cellulase.[Method] Aspergillus niger was used as the starting strain,and a high-yield production strain of cellulase was selected after UV mutation treatment.[Result] Under the suitable condition,the strain 2（15） with the highest CMC production capacity was selected,which nearly increased 50% than that of the starting strain.[Conclusion] The research provided the foundation for its appliation in the feed production in the future.

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.

Improvement of xylanase production by Aspergillus niger XY-1 using response surface methodology for optimizing the medium composition

Objective: To study the optimal medium composition for xylanase production by Aspergillus niger XY-1 in solid-state fermentation (SSF). Methods: Statistical methodology including the Plackett-Burman design (PBD) and the central composite design (CCD) was employed to investigate the individual crucial component of the medium that significantly affected the enzyme yield. Results: Firstly, NaNO3, yeast extract, urea, Na2CO3, MgSO4, peptone and (NH4)2SO4 were screened as the significant factors positively affecting the xylanase production by PBD. Secondly, by valuating the nitrogen sources effect, urea was proved to be the most effective and economic nitrogen source for xylanase production and used for further optimization. Finally, the CCD and response surface methodology (RSM) were applied to determine the optimal concentration of each sig-nificant variable, which included urea, Na2CO3 and MgSO4. Subsequently a second-order polynomial was determined by multiple regression analysis. The optimum values of the critical components for maximum xylanase production were obtained as follows: x1 (urea)=0.163 (41.63 g/L), x2 (Na2CO3)=?1.68 (2.64 g/L), x3 (MgSO4)=1.338 (10.68 g/L) and the predicted xylanase value was 14374.6 U/g dry substrate. Using the optimized condition, xylanase production by Aspergillus niger XY-1 after 48 h fermentation reached 14637 U/g dry substrate with wheat bran in the shake flask. Conclusion: By using PBD and CCD, we obtained the optimal composition for xylanase production by Aspergillus niger XY-1 in SSF, and the results of no additional expensive medium and shortened fermentation time for higher xylanase production show the potential for industrial utilization.

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.

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.

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.

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.

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.

Research on Pectase Secreted by Aspergillus Niger Degumming Kenaf Bast Fiber

In this paper, the aerobic metabolism mechanism of Aspergillus Niger （AS 3.3 50 ）, which is the most suitable bacteria for degumming kenaf fiber, is expounded, and macromolecular structure of pectin is also analyzed. The fracture position of the macromolecular chain of kenaf pectin and its outgrowth structure, affected by Endo-PG and Exo-PG are secreted by AS 3. 350 and explored in molecule scale. The optimal value of degumming parameters are fixed： temperature 34 - 36℃, time 48 - 50 h, pH 6.5 - 7.5. Compared with kenaf fibers obtained by natural method, the ones deguntmed by bio-enzymatic method possess of smoother surface, .better. Separation, less impairment and higher strength with. residual pectin percentage of 14.5 %.

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.

Biosorption of Cadmium by Fungal Biomass of Aspergillus niger

Objective To investigate the removal of cadmium from aqueous solution by waste fungal biomass of Aspergillus niger, originated from citric acid fermentation industry. Methods Batch adsorption test was used to study the biosorption equilibrium and isotherm. The Cd2+ concentration was measured with atomic adsorption spectrophotometer (AAS) HITACHI 180-80. Results The biosorption achieved equilibrium within 30 min. The adsorption isotherm could be described by Freundlich adsorption model, and the constants KF and 1/n were determined to be 2.07 and 0.18, respectively, and the correlation efficiency was 0.97. The optimal pH for Cd adsorption was 6.0. The cadmium-laden biomass could be effectively regenerated using 0.1 N HCl. Conclusion The waste biomass of Aspergillus niger, a by-product of fermentation industry, is a potential biosorbent for the removal of cadmium from aqueous solution.

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.

Production of Amylase Enzyme through Solid State Fermentation by Aspergillus niger

Traditionally, coconut dregs will be used as animal feed after the extraction of coconut oil and coconut milk from the copra. This study was carried out to discover the commercial value of coconut dregs as a solid substrate in the production of amylase through solid state fermentation （SSF） since this agro-waste is fairly rich in nutrients, providing the necessary nutrients supplementation for better microbial activity to produce enzymes. In this study, amylase is to be produced from coconut dregs by Aspergillus niger through solid state fermentation （SSF）. Three parameters were covered, which are incubation time, initial moisture content of substrate and inoculum sizes. SSF was carried out by using incubator at 37 ~C to test for enzyme activity at these following parameters： incubation time： 24, 48, 72, 96 and 120 hours; substrate moisture content： 64, 66, 68, 70 and 72% （w/w）; inoculum sizes： 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mL spore suspension （5.5 × 10^6 spores/mL）. Enzyme activities were measured through the estimation of liberated reducing sugars after the assay of amylase enzyme by using DNS （3, 5 dinitrosalicylic acid） method. Highest enzyme activities were obtained at these following parameters： incubation time： 72 hours （31.76 U/gds）; initial moisture content ofsubstrate： 66% （26.66 U / gds） and inoculum sizes： 2.0 mL （30.56 U/gds）.