This study aimed to identify the bioactive compounds of the ethyl acetate extract of Aspergillus niger SH2-EGY using GC-MS and to evaluate their protective role against aflatoxin B1 (AFB1)-induced oxidative stress, genotoxicity and cytotoxicity in rats. Six groups of male Sprague-Dawley rats were treated orally for 4 weeks included the control group, AFB1-treated group (80 μg/kg b.w); fungal extract (FE)-treated groups at low (140) or high dose (280) mg/kg b.w and the groups treated with AFB1 plus FE at the two tested doses. The GC-MS analysis identified 26 compounds. The major compounds found were 1,2,3,4,6-Penta-trimethylsilyl Glucopyranose, Fmoc-L-3-(2-Naphthyl)-alanine, D-(-)-Fructopyranose, pentakis (trimethylsilyl) ether, bis (2-ethylhexyl) phthalate, trimethylsilyl ether-glucitol, and octadecanamide, N-(2- methylpropyl)-N-nitroso. The in vivo results showed that AFB1 significantly increased serum ALT, AST, creatinine, uric acid, urea, cholesterol, triglycerides, LDL, carcinoembryonic antigen, alpha-fetoprotein, interleukin-6, Malondialdehyde, nitric oxide, Bax, caspase-3 and P53 mRNA expression, chromosomal aberrations and DNA fragmentation. It decreased serum TP, albumin, HDL, Bcl-2 mRNA expression, hepatic and renal TAC, SOD and GPx content and induced histological changes in the liver and kidney. FE prevented these disturbances in a dosage-dependent manner. It could be concluded that A. niger SH2-EGY extract is safe a promising agent for pharmaceutical and food industries.
The production of beta-mannanase from palm kernel cake (PKC) as a substrate in solid substrate fermentation (SSF) was studied using a laboratory column bioreactor. The simultaneous effects of three independent variables, namely incubation temperature, initial moisture content of substrate and airflow rate, on beta-mannanase production were evaluated by response surface methodology (RSM) on the basis of a central composite face-centered (CCF) design. Eighteen trials were conducted in which Aspergillus niger FTCC 5003 was cultivated on PKC in an aerated column bioreactor for seven days under SSF process. The highest level of beta-mannanase (2117.89 U/g) was obtained when SSF process was performed at incubation temperature, initial moisture level and aeration rate of 32.5 degrees C, 60% and 0.5 l/min, respectively. Statistical analysis revealed that the quadratic terms of incubation temperature and initial moisture content had significant effects on the production of beta-mannanase (P < 0.01). A similar analysis also demonstrated that the linear effect of initial moisture level and an interaction effect between the initial moisture content and aeration rate significantly influenced the production of beta-mannanase (P < 0.01). The statistical model suggested that the optimal conditions for attaining the highest level of beta-mannanase were incubation temperature of 32 degrees C, initial moisture level of 59% and aeration rate of 0.5 l/min. A beta-mannanase yield of 2231.26 U/g was obtained when SSF process was carried out under the optimal conditions described above.
Matched MeSH terms: Aspergillus niger/enzymology*; Aspergillus niger/growth & development
The production of xylanase from palm kernel cake as a substrate was studied in solid substrate fermentation. The simultaneous effects of three independent variables, namely incubation temperature, initial moisture content of substrate and air flow rate on xylanase production were evaluated by response surface methodology using central composite face centered design. A total of 18 experiments were carried out in which Aspergillus niger FTCC 5003 was cultivated on palm kernel cake in a column bioreactor for 7 days under incubation temperature, moisture level and aeration rate determined. Test results showed that the highest xylanase activity of 174.88 U g(-1) was produced at incubation temperature, initial moisture level and aeration rate of 25 degrees C, 60% and 1.5 L min(-1), respectively. The statistical analysis of the experimental results revealed that the linear effect of incubation temperature and quadratic term of initial moisture content had highly significant effects on xylanase production (p<0.01). Statistical results also showed that interaction effect between incubation temperature and initial moisture content as well as interaction effect between moisture level and aeration rate influenced the yield ofxylanase at probability levels of 95%. Optimum conditions determined by statistical model for attaining maximum xylanase production were incubation temperature of 25 degrees C, initial moisture level of 63% and aeration rate of 1.76 L min(-1). The xylanase activity of 192.50 U g(-1) was obtained when solid substrate fermentation was performed under the optimal circumstances.
The leaves of Gynotroches axillaris were chemically and biologically studied. Sequential extraction of the leaves using petroleum ether, chloroform, and methanol afforded three extracts. Purification of pet. ether extract yielded, squalene and β-amyrin palmitate as the major compounds, together with palmitic acid and myristic acid as the minor components. The methanol extract yielded two flavonoids, quercitrin and epicatechin. The isolated compounds were characterized by MS, IR and NMR (1D and 2D). Anti-acetyl cholinesterase screening using TLC bio-autography assay showed that palmitic acid and myristic acid were the strongest inhibition with detection limit 1.14 and 1.28 μ/g/ 5 μL respectively. Antibacterial against Gram-positive and negative and antifungal activities exhibited that β-amyrin palmitate was the strongest (450-225 μ/mL) against all the tested microbes. The tyrosinase inhibition assay of extracts and the pure compounds were screened against tyrosinase enzyme. The inhibition percentage (I%) of methanol extract against tyrosinase enzyme was stronger than the other extracts with value 68.4%. Quercitrin (59%) was found to be the highest in the tyrosinase inhibition activity amongst the pure compounds. To the best of our knowledge, this is first report on the phytochemicals, tyrosinase inhibition, anti-acetycholinesterase and antimicrobial activities of the leaves of G. axillaris.
Matched MeSH terms: Aspergillus niger/drug effects; Aspergillus niger/growth & development
A study on liquid state bioconversion of sewage treatment plant (STP) sludge was assisted to evaluate the performance of batch fermenter compared to shake flask in a laboratory. Bioconversion of STP sludge was highly influenced by the mixed fungal culture of Penicillium corylophilum and Aspergillus niger after 4 days of treatment. The results showed that about 24.9 g kg(-1) dry sludge cake (DSC) was produced with enrichment of fungal biomass protein in fermenter while 20.1 g kg(-1) in shake flask after 4 days of fungal treatment. The effective biodegradation of STP sludge was recorded in both fermenter and shake flask experiment compared to control (uninnoculated sample). The results presented in this study revealed that the overall performance of fermenter in terms of sludge cake (biosolids) accumulation and biodegradation of STP sludge was higher than the shake flask.
The bioconversion of domestic wastewater sludge by immobilized mixed culture of filamentous fungi was investigated in a laboratory. The potential mixed culture of Penicillium corylophilum WWZA1003 and Aspergillus niger SCahmA103 was isolated from its local habitats (wastewater and sludge cake) and optimized on the basis of biodegradability and dewaterability of treated sludge. The observed results in this study showed that the sludge treatment was highly influenced by the effect of immobilized mixed fungi using liquid state bioconversion (LSB) process. The maximum production of dry filter cake (DFC) was enriched with fungal biomass to about 20.05 g/kg containing 23.47 g/kg of soluble protein after 4 days of fungal treatment. The reduction of COD, TSS, turbidity (optical density against distilled water, 660 nm), reducing sugar and protein in supernatant and filtration rate of treated sludge were influenced by the fungal mixed culture as compared to control (uninnoculated). After these processes, 99.4% of TSS, 98.05% of turbidity, 76.2% of soluble protein, 98% of reducing sugar and 92.4% of COD in supernatant of treated sludge were removed. Filtration time was decreased tremendously by the microbial treatment after 2 days of incubation. The effect of fungal strain on pH was also studied and presented. Effective bioconversion was observed after 4 days of fungal treatment.
Effects of agitation and aeration rate on microbial treatment of domestic wastewater sludge were investigated in a batch fermenter using mixed culture of Penicillium corylophilum and Aspergillus niger. It was found that liquid state bioconversion (LSB) of wastewater sludge was highly influenced by the effects of agitation and aeration. The maximum production of sludge cake and reduction of organic substances in treated sludge were recorded at 150-200 rpm of agitation speed and 0.5 vvm of aeration rate after 72 h of treatment. No effective results were observed at higher rate of agitation (300 rpm) and aeration (1.5 vvm) as compared to optimum values. The results showed that the minimum level of air saturation (pO2) was adequate to maintain the bioconversion process.
The biosolids accumulation and biodegradation of domestic wastewater treatment plant (DWTP) sludge by filamentous fungi have been investigated in a batch fermenter. The filamentous fungi Aspergillus niger and Penicillium corylophilum isolated from wastewater and DWTP sludge was used to evaluate the treatment performance. The optimized mixed inoculum (A. niger and P. corylophilum) and developed process conditions (co-substrate and its concentration, temperature, initial pH, inoculum size, and aeration and agitation rate) were incorporated to accelerate the DWTP sludge treatment process. The results showed that microbial treatment of higher strength of DWTP sludge (4% w/w of TSS) was highly influenced by the liquid state bioconversion (LSB) process. In developed bioconversion processes, 93.8 g/kg of biosolids was enriched with fungal biomass protein of 30 g/kg. Enrichment of nutrients such as nitrogen (N), phosphorous (P), potassium (K) in biosolids was recorded in 6.2% (w/w), 3.1% (w/w) and 0.15% (w/w) from its initial values of 4.8% (w/w), 2.0% (w/w) and 0.08% (w/w) respectively after 10 days of fungal treatment. The biodegradation results revealed that 98.8% of TSS, 98.2% of TDS, 97.3% of turbidity, 80.2% of soluble protein, 98.8% of reducing sugar and 92.7% of COD in treated DWTP sludge supernatant were removed after 8 days of microbial treatment. The specific resistance to filtration (SRF) in treated sludge (1.4x10(12) m/kg) was decreased tremendously by the microbial treatment of DWTP sludge after 6 days of fermentation compared to untreated sample (85x10(12) m/kg).
A study was conducted to evaluate the settleability and dewaterability of fungal treated and untreated sludge using liquid state bioconversion process. The fungal mixed culture of Aspergillus niger and Penicillium corylophilum was used for fungal pretreatment of wastewater sludge. The fungal strains immobilized/entrapped on sludge particles with the formation of pellets and enhanced the separation process. The results presented in this study showed that the sludge particles (pellets) size of 2-5mm of diameter were formed with the microbial treatment of sludge after 2 days of fermentation that contained maximum 33.7% of total particles with 3-3.5mm of diameter. The settling rate (measured as total suspended solids (TSS) concentration, 130 mg/l) was faster in treated sludge than untreated sludge (TSS concentration, 440 mg/l) after 1 min of settling time. In 1 min of settling operation, 86.45% of TSS was settled in treated sludge while 4.35% of TSS settled in raw sludge. Lower turbidity was observed in treated sludge as compared to untreated sludge. The results to specific resistance to filtration (SRF) revealed that the fungal inoculum had significant potentiality to reduce SRF by 99.8% and 98.7% for 1% w/w and 4% w/w of TSS sludge, respectively. The optimum fermentation period recorded was 3 days for 1% w/w sludge and 6 days for 4% w/w sludge, respectively, for dewaterability test.
This study evaluated the potential of bioflocculant production from Aspergillus niger using palm oil mill effluent (POME) as carbon source. The bioflocculant named PM-5 produced by A. niger showed a good flocculating capability and flocculating rate of 76.8% to kaolin suspension could be achieved at 60 h of culture time. Glutamic acid was the most favorable nitrogen source for A. niger in bioflocculant production at pH 6 and temperature 35 °C. The chemical composition of purified PM-5 was mainly carbohydrate and protein with 66.8% and 31.4%, respectively. Results showed the novel bioflocculant (PM-5) had high potential to treat river water from colloids and 63% of turbidity removal with the present of Ca(2+) ion.
Citric acid (CA) has a high demand due to its various uses in the food and pharmaceutical industries. However, the natural supply of CA is minimal compared to its growing industrial demand. The increasing demand for CA can be fulfilled by using biotechnological processes. This study utilized liquid state bioconversion by Aspergillus niger for CA production using sugarcane molasses as the primary substrate. Sugarcane molasses which is agricultural waste consists of significant proportion of organic matters such as lipids and carbohydrates. This makes sugarcane molasses as a potential and alternative source of producing CA at a lower cost. In this study, statistical optimization was applied to improve CA production using submerged fermentation in shake flasks. Aspergillus niger was cultured in potato dextrose agar. Then, inoculum spores were introduced into the fermentation media for a specific duration according to the experimental design from Central Composite Design (CCD) tool under Response Surface Methodology (RSM) in Design Expert 6.0 software. Three parameters were chosen to be optimized at 32⁰C i.e.agitation rate (160, 80, 200 rpm), substrate concentration (47, 60, 73%) and fermentation time (24, 72, 120 h). High Performance Liquid Chromatography (HPLC)and Fourier-transform infrared spectroscopy(FTIR) analyses were conducted to measure CA yield. The optimization study showed that the media incubated for 72 hours with a substrate concentration of 60% and an agitation speed of 180 rpm produced the highest CA yield(21.2 g/L).The analysis of variance (ANOVA) also showed that CCD quadratic model was significant with P-value< 0.0104 and R2is0.8964.
A. niger produced alpha-glucosidase, alpha-amylase and two forms of glucoamylase when grown in a liquid medium containing raw tapioca starch as the carbon source. The glucoamylases, which formed the dominant components of amylolytic activity manifested by the organism, were purified to homogeneity by ammonium sulfate precipitation, ion-exchange and two cycles of gel filtration chromatography. The purified enzymes, designated GA1 and GA2, a raw starch digesting glucoamylase, were found to have molar masses of 74 and 96 kDa and isoelectric points of 3.8 and 3.95, respectively. The enzymes were found to have pH optimum of 4.2 and 4.5 for GA1 and GA2, respectively, and were both stable in a pH range of 3.5-9.0. Both enzymes were thermophilic in nature with temperature optimum of 60 and 65 degrees C, respectively, and were stable for 1 h at temperatures of up to 60 degrees C. The kinetic parameters Km and V showed that with both enzymes the branched substrates, starch and amylopectin, were more efficiently hydrolyzed compared to amylose. GA2, the more active of the two glucoamylases produced, was approximately six to thirteen times more active towards raw starches compared to GA1.
An amperometric enzyme-electrode was introduced where glucose oxidase (GOD) was immobilized on chitosan membrane via crosslinking, and then fastened on a platinum working electrode. The immobilized enzyme showed relatively high retention activity. The activity of the immobilized enzyme was influenced by its loading, being suppressed when more than 0.6 mg enzyme was used in the immobilization. The biosensor showing the highest response to glucose utilized 0.21 ml/cm2 thick chitosan membrane. The optimum experimental conditions for the biosensors in analysing glucose dissolved in 0.1 M phosphate buffer (pH 6.0) were found to be 35°C and 0.6 V applied potential. The introduced biosensor reached a steady-state current at 60 s. The apparent Michaelis-Menten constant ([Formula: see text]) of the biosensor was 14.2350 mM, and its detection limit was 0.05 mM at s/n > 3, determined experimentally. The RSD of repeatability and reproducibility of the biosensor were 2.30% and 3.70%, respectively. The biosensor was showed good stability; it retained ~36% of initial activity after two months of investigation. The performance of the biosensors was evaluated by determining the glucose content in fruit homogenates. Their accuracy was compared to that of a commercial glucose assay kit. There was no significance different between two methods, indicating the introduced biosensor is reliable.
This study was aimed at identifying indigenous microorganisms from palm oil mill effluent (POME) and to ascertain the microbial load. Isolation and identification of indigenous microorganisms was subjected to standard microbiological methods and sequencing of the 16S rRNA and 18S rRNA genes. Sequencing of the 16S rRNA and 18S rRNA genes for the microbial strains signifies that they were known as Micrococcus luteus 101PB, Stenotrophomonas maltophilia 102PB, Bacillus cereus 103PB, Providencia vermicola 104PB, Klebsiella pneumoniae 105PB, Bacillus subtilis 106PB, Aspergillus fumigatus 107PF, Aspergillus nomius 108PF, Aspergillus niger 109PF and Meyerozyma guilliermondii 110PF. Results revealed that the population of total heterotrophic bacteria (THB) ranged from 9.5 × 105 - 7.9 × 106 cfu/mL. The total heterotrophic fungi (THF) ranged from 2.1 × 104 - 6.4 × 104 cfu/mL. Total viable heterotrophic indigenous microbial population on CMC agar ranged from 8.2 × 105 - 9.1 × 106 cfu/mL and 1.4 × 103 - 3.4 × 103 cfu/mL for bacteria and fungi respectively. The microbial population of oil degrading bacteria (ODB) ranged from 6.4 × 105 - 4.8 × 106 cfu/mL and the oil degrading fungi (ODF) ranged from 2.8 × 103 - 4.7 × 104 cfu/mL. The findings revealed that microorganisms flourish well in POME. Therefore, this denotes that isolating native microorganisms from POME is imperative for effectual bioremediation, biotreatment and biodegradation of industrial wastewaters.
A sequential optimization based on statistical design and one-factor-at-a-time (OFAT) method was employed to optimize the media constituents for the improvement of citric acid production from oil palm empty fruit bunches (EFB) through solid state bioconversion using Aspergillus niger IBO-103MNB. The results obtained from the Plackett-Burman design indicated that the co-substrate (sucrose), stimulator (methanol) and minerals (Zn, Cu, Mn and Mg) were found to be the major factors for further optimization. Based on the OFAT method, the selected medium constituents and inoculum concentration were optimized by the central composite design (CCD) under the response surface methodology (RSM). The statistical analysis showed that the optimum media containing 6.4% (w/w) of sucrose, 9% (v/w) of minerals and 15.5% (v/w) of inoculum gave the maximum production of citric acid (337.94 g/kg of dry EFB). The analysis showed that sucrose (p<0.0011) and mineral solution (p<0.0061) were more significant compared to inoculum concentration (p<0.0127) for the citric acid production.
Nanobiotechnology has emerged as a promising technology to develop new therapeutically active nanomaterials. The present study was aimed to biosynthesize AgNPs extracellularly using Aspergillus niger JX556221 fungal extract and to evaluate their anticancer potential against colon cancer cell line, HT-29. UV-visible spectral characterization of the synthesized AgNPs showed higher absorption peak at 440 nm wavelength. Transmission Electron Microscopy (TEM) analysis revealed the monodispersed nature of synthesized AgNPs occurring in spherical shape with a size in the range of 20-25 nm. Further, characterization using Energy Dispersive Spectroscopy (EDX) confirmed the face-centred cubic crystalline structure of metallic AgNPs. FTIR data revealed the occurrence of various phytochemicals in the cell free fungal extract which substantiated the fungal extract mediated AgNPs synthesis. The cytotoxic effect of AgNPs was studied by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results evidenced the cytotoxic effect of AgNPs on HT-29 cell lines in a dose dependent manner. The highest activity was found at 100 μg/ml concentration after 24 h of incubation. Use of propidium iodide staining examination method confirmed the cytotoxic effect of AgNPs through inducing cell apoptosis. AgNPs cytotoxicity was found to be through elevating reactive oxygen species (ROS), and caspase-3 activation resulting in induced apoptosis. Therefore, this research finding provides an insight towards the development of novel anticancer agents using biological sources.
Kajian keberkesanan sifat antimikrob ekstrak kacangma dijalankan menggunakan ujian resapan cakera dan ujian perencatan langsung. Hasil menunjukkan ekstrak etanol dengan kepekatan 50 dan 100 mg/mL merencat Staphylococcus aureus. Bagi ekstrak air, kepekatan 10, 25, 50 dan 100 mg/mL merencat Aspergillus niger, 25, 50 dan 100 mg/mL dapat merencat Saccharomyces cerevisae dan kepekatan 100 mg/mL dapat merencat Staphylococcus aureus. Perlakuan suhu yang berbeza ke atas ekstrak dalam ujian perencatan langsung tidak menunjukkan sebarang perbezaan ke atas perencatan mikroorganisma yang dikaji.
Agitation speed was found to influence the tannase production and fungal growth of Aspergillus niger FETL FT3. The optimal agitation speed was at 200 rpm which produced 1.41 U/ml tannase and 3.75 g/l of fungal growth. Lower or higher agitation speeds than 200 rpm produced lower enzyme production and fungal growth. Based on the SEM and TEM micrograph observation, there was a significant correlation between agitation speed and the morphology of the fungal mycelia. The results revealed an increase of the enzyme production with the change of the fungal growth morphology from filamentous to pelleted growth forms. However, the exposure to higher shear stress with an increasing agitation speed of the shaker also resulted in lower biomass yields as well as enzyme production.
The ability of immobilized cell cultures of Aspergillus niger FETL FT3 to produce extracellular tannase was investigated. The production of enzyme was increased by entrapping the fungus in scouring mesh cubes compared to free cells. Using optimized parameters of six scouring mesh cubes and inoculum size of 1 × 10(6) spores/mL, the tannase production of 3.98 U/mL was obtained from the immobilized cells compared to free cells (2.81 U/mL). It was about 41.64% increment. The immobilized cultures exhibited significant tannase production stability of two repeated runs.
Hevea brasiliensis extract could potentially be employed as a relatively low cost resource for various anti-fungal activities due to the simplicity of latex preparation and the abundance of latex that can be obtained in rubber producing regions. The present study was aimed at examining the species specific anti-fungal property of H. brasilensis latex C-serum against Aspergillus niger.
Matched MeSH terms: Aspergillus niger/drug effects*; Aspergillus niger/growth & development