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.

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.

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.

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.

响应面法优化Aspergillus niger X-6发酵生产菊粉酶工艺的研究

为提高Aspergillus niger X-6发酵生产菊粉酶的产率,运用Plackett-Burmen方法对麸皮、菊粉、蛋白胨、酵母膏添加量和发酵时间、温度、pH值、接种量8个影响因素进行考察,筛选出麸皮添加量、发酵时间和pH值为3个显著影响因素,然后采用Box-Behnken中心组合和响应面法对上述3个因素进行产酶条件的进一步优化,建立菊粉酶产率的二次多项式数学模型,并分析模型的有效性与因子间的交互作用。结果表明:黑曲霉发酵产菊粉酶的优化工艺参数为:麸皮添加量4.64%、发酵时间81.5h、pH6.0。在此条件下,产酶活力达20.42U/mL,与优化前相比提高了30.81%。

黑曲霉Aspergillus niger木质纤维素降解能力及产酶研究

从农林废物堆肥中分离得到一株真菌经鉴定为黑曲霉,将该菌用于木质素类化合物利用,固态培养条件下考察其对木质纤维素的降解能力及产酶特性,另外对发酵前后的稻草结构进行了红外光谱分析。结果表明,黑曲霉具有木质素降解能力,兼具低分子量木质素酚型、非酚型类物质的降解能力。其对木质素降解是木质素过氧化物酶、锰过氧化物酶、漆酶、纤维素酶和半纤维素酶共同作用的结果,黑曲霉产酶高峰处于菌体生长较稳定的时期。在本实验条件下,培养30d使木质素降解率达16.87%,同时对纤维素、半纤维素也有较高程度的降解;降解率分别为39.85%、45.32%。红外光谱分析结果表明,稻草木质素结构被破坏,黑曲霉木质素各官能团的降解作用有所不同。

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.

黑曲霉Aspergillus niger SL2-111所产酸性蛋白酶的分离纯化及酶学特性

采用硫酸铵盐析、Sephadex G-25柱脱盐、DEAE—Sepharose Fast Flow离子交换层析、Sephacryl S-200分子筛层析和高效液相色谱等方法，从黑曲霉菌株Aspergillus niger SL2—111的发酵曲中提取了一种酸性蛋白酶，经SDS—PAGE验证，该酶纯化水平已达到电泳纯．该酶的表观分子量（Mr）约为47×10^3，最适pH值为3．0，pH稳定性范围为2．5—6．0，最适温度为50℃，温度稳定性范围为30—60℃；Cu^2＋、Mn^2＋对其有激活作用，Hg^2＋、Ag^＋对其则有轻度的抑制作用；该酶的氨基酸组成为：中极性酸性氨基酸占17．29％，极性碱性氨基酸占4．50％，极性中性氨基酸占38．50％，其它为非极性氨基酸；N端氨基酸序列为SKGSAVTFPQ，经序列同源性比对，表明该酶与其它曲霉酸性蛋白酶具有极高的同源性．

生淀粉糖化酶产生菌Aspergillus niger(6#)的分离筛选及其产酶条件

从大曲中分离到了1株降解生淀粉能力较强的黑曲霉Aspergillusniger(6#).固体发酵酶活可达2461U/g,RDA值为21.47%;液体发酵酶活可达353U/mL,RDA值为20.30%.以6#菌为实验材料,进一步考察了氮源、碳源及pH对生淀粉糖化酶形成的影响.实验结果表明,无机氮源NaNO3较有机氮源蛋白胨更有利于生淀粉糖化酶的形成;pH是影响生淀粉糖化酶形成的重要因素,低pH会阻遏生淀粉糖化酶的形成;玉米粉和麦芽糖较其它碳源更有利于生淀粉糖化酶的形成.

黑曲霉(Aspergillus niger)产β-葡聚糖酶固态发酵优化的研究

研究在黑曲霉 (Asp niger) FSN6 5固态发酵中碳氮比、无机氮源、大麦粉添加、水分比例、初始 pH、接种量、培养温度及发酵时间对β 葡聚糖酶酶产量的影响。结果表明 ,培养基中C/N(以麸皮与豆饼粉比例计 )为 8∶1;最佳无机氮源为NH4 NO3;大麦添加对产酶没有明显的诱导作用 ;培养基中最适水分比例为 1∶1;最适发酵条件 :初始发酵pH为 6 0 ;最适接种量为每瓶 0 5mL孢子悬液 (孢子浓度为 4 5× 10 7/mL) ;最适的发酵温度为 33℃ ;在以上最适条件下固态培养 70h ,发酵产酶水平可达 14 16 49u/ g ,优化结果比初始设计提高了 2 6 %。对粗酶酶学特性研究表明 :该酶最适作用 pH为 5 0 ,最适作用温度为 75℃。

黑曲霉(Aspergillus niger)产菊粉酶发酵条件的研究

以菊粉为唯一碳源,对样土进行产菊粉酶黑曲霉菌株的初筛,得到17株菌株。用跟踪测酶活的方法进行复筛,获得3株高产菊粉酶菌株,最高酶活达到25.41U·mL-1。然后探讨了不同碳源和氮源对黑曲霉产菊粉酶活力的影响,结果表明,最佳碳源为菊粉,氮源为酵母膏。通过正交实验,得到了优化的产菊粉酶黑曲霉液体发酵条件:菊粉用量为2%,pH5.0,250mL三角瓶中装培养基量为50mL,发酵温度为30℃。

黑曲霉(Aspergillus niger)纤维素酶系中内切β-葡聚糖酶性质的研究

对黑曲霉菌株Ｗ２５纤维素酶系中内切β－萄聚糖酶组分的性质进行了研究。Ⅰ－２ａ，Ⅰ－２ｂ，Ⅰ－２ｃ，Ⅰ－２ｄ，Ⅱａ，Ⅱｂ，和Ⅲａ的最适ｐＨ分别为５．５，５．５，５．０，５．５，５．０，５．５和５．５；均在ｐＨ２．０－７．０之间稳定，最适温度分别为５０，５０，５５，５５，５５和６０℃；保温１小时的半失活温度ｔ１／２分别为４９、６１、６３、５２、５７、４７和６７℃。ＳＤＳ－凝胶电脉法测得Ⅰ－２ａ。

低能离子注入木聚糖酶产生菌黑曲霉(Aspergillus Niger)育种中参数的优化

本文对离子束技术的注入工艺和在不同能量下的不同注入剂量下黑曲霉的存活率和突变率进行了研究,目的是为了找到恰当的注入工艺和良好的注入参数,使诱变达到较好的效果。结果发现:当注入离子为氮离子时,在10keV的能量下注入剂量为5.2×1014ions/cm2和1.56×1015ions/cm2时,诱变效果较好。在这注入条件下,通过培养基优化,使木聚糖酶酶活达到600IU/ml。

果胶酶高产菌株Aspergillus niger3502的选育

采用紫外线和微波处理黑曲霉 (Aspergillusniger)孢子 ,获得 1株比出发菌株果胶酶产生能力提高 2倍的突变株 35 0 2 ;在用正交试验优选而得的最佳固体发酵条件下 ,酶活力达 2

黑曲霉(Aspergillus niger)产菊粉酶菌株的筛选及培养条件的研究

从3个地区7个土壤样品中筛选高产菊粉酶的黑曲霉菌株,对筛选出的黑曲霉菌株进行了形态鉴定,探讨了温度、pH值对菊粉酶活力的影响,比较了固体和液体两种扩大培养法对黑曲霉菌株产菊粉酶的影响。结果表明:经过初筛和复筛得到A4、A6、A8、A12、A14、A15和A16共7株黑曲霉菌株,菊粉酶活力大于15.0 U/mL,经数码生物摄影显微镜镜检符合黑曲霉的形态特征。A8菌株的菊粉酶酶活最高,达到25.0U/mL,在60℃、pH值5.0的条件下酶活性最大。

Aspergillus niger FS6中β-1,3-1,4-葡聚糖酶cDNA片段的克隆及序列测定

通过 RT- PCR方法首次从真菌 Aspergillus niger FS6中扩增和克隆到 β- 1,3- 1,4 -葡聚糖酶 c DNA片段 .序列测定表明 :该片段全长 2 80 bp.经 Clustal W软件进行同源性分析后表明 ,该序列与枯草芽孢杆菌、水解淀粉芽孢杆菌、厌氧真菌、牛链球菌和热纤维梭菌中 β- 1,3- 1,4 -葡聚糖酶基因同源性很高 ,分别为 91% ,90 % ,70 % ,6 7% ,6 5 %

磷矿石的微生物风化作用——以一株黑曲霉(Aspergillus niger)为例

以直接和间接作用的方式研究培养基中黑曲霉Aspergillus niger对磷矿粉的风化作用。在装有100目磷矿石粉的液体培养基中接入该菌研究其对磷矿石粉的直接风化作用;同时将装有100目磷矿石粉的透析袋放入液体培养基中再接入该菌研究其对磷矿石粉的间接风化作用。按不同时间取培养液上清液,用电感耦合等离子体-发射光谱仪测定Ca2+、Mg2+、Al3+、Fe3+、K+和Mn2+浓度,用离子色谱法测定H2PO4-、SO42-和Cl-浓度。此外,黑曲霉风化作用后的矿物残渣用电子探针作表面微观形态分析和XRD矿物物相分析。结果表明:黑曲霉对磷矿石粉风化作用的直接作用强度大于间接作用;黑曲霉可风化磷矿石粉,但对矿粉中不同矿物的风化强度不同,风化过程中形成次生矿物水草酸钙。提出黑曲霉对磷矿石的风化作用源自黑曲霉生长导致的机械破坏作用、胞外分泌物的生化降解作用以及多种因素之间的协同作用。

黑曲霉U_γ-2(Aspergillus niger U_γ-2)产菊粉酶的研究

研究了黑曲霉Uγ- 2的产酶进程及其产酶条件。结果表明 :在 1g·mL-1菊芋汁中添加 30mg·mL-1的牛肉膏 ,调节初始pH 6 0 ,摇瓶转速为 180r·min-1,30℃条件下发酵最高酶活力为 180 33μmol·min-1·mL-1。发酵初期产酶速率较快 ,pH下降速度快。在发酵进行到 72～ 96h期间 ,出现产酶的迟缓期 ,此时发酵液的pH为 4 6 2～ 4 38;以后菊粉酶活力明显上升。在 192h时菊粉酶活力达到最高值 ,发酵液pH降到最低点。在发酵进程中 ,发酵液可溶性蛋白出现 2个高峰 ,72h时出现第一高峰 ,与菊粉酶活力比较表明此时主要是杂蛋白产生所致 ;在 192h时出现的第二高峰与菊粉酶活力同步 ,说明主要是菊粉酶产生。与生物量的变化曲线比较 ,尽管在发酵初期产酶速率较快 ,但菊粉酶在发酵液中大量积累出现在 12 0h后 (对数增长期的后期 )

Aspergillus niger S-05产柚苷酶液态发酵条件的优化

通过单次单因素试验对黑曲霉(Aspergillus niger S-05)液体发酵的产柚苷酶条件进行优化,结果表明,当培养条件为豆粉2.0%、蔗糖0.5%、柚苷0.05%、MgSO4·7H2O0.6%、吐温-800.2%、培养温度26℃、初始pH5.0、250mL三角瓶装量30mL、转速190r/min、培养时间105h时柚苷酶酶活达到最高值(429.09U/mL)。同时发现,通过调节培养基组成可改变所产柚苷酶中α-L-鼠李糖苷酶和β-D-葡萄糖苷酶的酶活比例,从而实现根据不同应用进行有针对性的发酵产酶控制目的。