Microbial lipase plays a crucial role in industrial biotechnology by contributing to a wide range of industrial products worldwide. Seeking potential lipase producers and enhancing their productivity represent a current urge. This study focuses on optimizing the culture conditions required for the extracellular production of lipase by Aspergillus niger isolates through submerged fermentation. Aspergillus niger was isolated from different herbal seeds and oily sources. Seven fungal isolates could be morphologically identified by the Assiut University Mycological Center (AUMC) and were cultivated on Czapek's Dox Medium. Various pH values, temperatures, oil concentrations, and metal ion (Mn2+, Zn2+, Cu2+ and Mg2+) concentrations were investigated to assess their impact on isolates growth, total extracellular protein concentration, and lipase activity. For the isolate AUMC 16212, optimum lipase production was reached at (pH 5 / 37 degrees C / 3% corn oil / 10 mM Mn2+ / 10 mM Cu2+ / 2 mM Zn2+ / 22 mM Mg2+). While that from AUMC 16213 isolate was (pH 5 / 37 degrees C / 3% corn oil/ 8 mM Mn2+ / 10 mM Cu2+ / 2 mM Zn2+ / 22 mM Mg2+) The culture conditions of two isolates of Aspergillus niger (AUMC 16212 and 16213) could be optimized for potential lipase production which is of ascending industrial and economical value.

Metalaxyl (MY) is a water soluble synthetic acylanilide fungicide widely used in agriculture practices to control phytopathogenic fungi, but it elicits environmental pollution to underground water as well as human health. The safe and effective removal of agrochemical pollutants from contaminated field sites is performed by some soil and aquatic microorganisms. However, limited work has been done focusing on the intermediate metabolites' residues produced and enzymes involved in the biodegradation process of fungicides. The degradation ability of the isolated aquatic fungus Aspergillus wentii of the MY as a sole carbon source in only sterilized distilled water was evaluated in vitro after 15 days, reaching high levels of biodegradation at 68%. Furthermore, the possible catabolic degradation pathways of MY based on the detected metabolites beside the enzymes involved in the biodegradation process were established. Moreover, a TripleTOF 5600 + LC-MS/MS was employed to identify the enzymes involved in the biodegradation process of the fungicide. The results revealed that CYP450 is responsible for metabolism of MY. It could be concluded that Aspergillus wentii enzymes have a prospective potential to eliminate a wide spectrum of toxic environmental pollutants, such as MY. To our knowledge, this study is the first report that Aspergillus wentii CYP 450 could catalyze the degradation of metalaxyl.

Bioelectricity is considered one of the most important sources of clean green energy that has attracted the attention of many scientists to discover some of its secrets. Moreover, agro-industrial wastes are widely used in the production of alternative energy in general and electricity in particular. There is an important agro-industrial waste produced during the molasses (black honey) industry in Upper Egypt, that waste is called (El-ghasheem). In the present work, El-ghasheem was used in an economical-designed microbial fuel cell (MFC) to obtain cheap green electricity. The 'H'-type MFC was operated with a capacity of 600 ml in each chamber, and copper wire was used to link the two graphite rod electrodes. Lamp wicks that had been soaked in a 1M KCl solution were used to create an inexpensive salt bridge. Factors affecting the production of bioelectricity from this MFC were optimized to get the optimum power. B ioelectricity production was enhanced using crude microbial laccase enzyme as a cathodic reaction biocatalyst. Laccase produced from Penicillium chrysogenum was used to increase voltage and current from 0.40 +/- 0.011 V, 0.02 +/- 0.002 A to 0.608 +/- 0.002 V and 0.05 +/- 0.003 A, respectively at 37 degrees C, anolyte pH 6, and catholyte pH 5 for a 10-day incubation period.

In this study, marine Streptomyces exfoliatus was isolated from the Red Sea, Egypt, and was identified using 16s rRNA.S. exfoliatus was fermented on different media under different carbon source concentrations, pH, temperatures, distilled H 2 O, and sea H 2 O, under shaking and static conditions.The active material was isolated by chloroform and ethyl acetate extractions then separated by column chromatography and purified by preparative TLC and identified by GC-MS.Methicillin-resistant Staphylococcus aureus inhibited by ß-caryophyllene.

Acinetobacter baumannii is a bacterial pathogen, associated with hospital and community acquired infections. It is one of the most common, opportunistic, and serious multidrug resistant bacteria (MDR). It is considered a global problem. The use of natural compounds such as fungal metabolites as an alternative to antibiotics is being explored to overcome the issue of antimicrobial resistance. Among fifty fungal spp. isolated from different sources, only Aspergillus terreus showed an antibacterial effect against A. baumannii. Co-culture is applied to produce bioactive compounds in higher amounts and/or to induce the production of new secondary metabolites. Several solvents and chromatographic techniques were used to isolate, fractionate, and purify Aspergillus terreus/ Acinetobacter baumannii co-culture secondary metabolites. Ethyl acetate extract was the most potent extract against MDR A. baumannii, in comparison to chloroform and methanol extracts. Characterization and structure elucidation of the purified compound isolated from ethyl acetate extract were performed using LC-MS, 1H NMR, and IR. The compound was identified as terreusinone A with a minimum inhibitory concentration of 160 µg.µl-1against MDR A. baumannii. This is the first record to determine the antibacterial effect of terreusinone A particularly against MDR A. baumannii.

Endobiotic fungi are considered as a reservoir of numerous active metabolites. Asparaginase is used as an antileukemic drug specially to treat acute lymphoblastic leukaemia. The presented study aims to optimize the media conditions, purify, characterize, and test the antileukemic activity of the asparaginase induced from Lasiodiplodia theobromae. The culture medium was optimized using an experiment designed by The Taguchi model with an activity ranging from 10 to 175 IU/mL. Asparaginase was induced with an activity of 315 IU/mL. Asparaginase was purified with a specific activity of 468.03 U/mg and total activity of 84.4 IU/mL. The purified asparaginase showed an approximate size of 70 kDa. The purified asparaginase showed an optimum temperature of 37 °C and an optimum pH of 6. SDS reduced the activity of asparaginase to 0.65 U/mL while the used ionic surfactants enhanced the enzyme activity up to 151.92 IU/mL. The purified asparaginase showed a Km of 9.37 µM and Vmax of 127.00 µM/mL/min. The purified asparaginase showed an IC50 of 35.2 ± 0.7 IU/mL with leukemic M-NFS-60 cell lines and CC50 of 79.4 ± 1.9 IU/mL with the normal WI-38 cell line. The presented study suggests the use of endophytic fungi as a sustainable source for metabolites such as asparaginase, provides an opportunity to develop a facile, eco-friendly, cost-effective, and rapid synthesis of antileukemic drugs, which have the potential to be used as alternative and reliable sources for potent anticancer agents.

Fungal–bacterial co-culturing is a potential technique for the production of secondary metabolites with antibacterial activity. Twenty-nine fungal species were screened in a co-culture with carbapenem-resistant Klebsiella pneumoniae at different temperatures. A temperature of 37 ° showed inhibition of bacterial growth. Antimicrobial susceptibility testing for K. pneumoniae was conducted to compare antibiotic resistance patterns before and after the co-culture. Genotypic comparison of the K. pneumonia was performed using next generation sequencing (NGS). It was shown that two out of five K. pneumoniae, with sequence type ST 101 isolates, lost bla-OXA48, bla-CTX-M-14, tir, strA and strB genes after the co-culture with Scopulariopsis brevicaulis fungus. The other three isolates (ST 383 and 147) were inhibited in the co-culture but did not show any changes in resistance. The total ethyl acetate extract of the fungal–bacterial co-culture was tested against K. pneumoniae using a disc diffusion method. The concentration of the crude extract was 0.97 mg/µL which resulted in total inhibition of the bacteria. Using chromatographic techniques, the purified compounds were identified as 11-octadecenoic acid, 2,4-Di-tert-butylphenol, 2,3-Butanediol and 9-octadecenamide. These were tested against K. pneumoniae using the well diffusion method at a concentration of 85 µg/µL which resulted in total inhibition of bacteria. The co-culture results indicated that bacteria under chemical stress showed variable responses and induced fungal secondary metabolites with antibacterial activities.

Sequancing of pullulanase from the fungus Aureobasidium pullulans isolated from Egypt soil; Genomic DNA of pullulanase was determined for the first time using PCR, according to Baser program, Pullulanase nucleotide collection from Aureobasidium pullulans was blasted which showed similarity using NCBI significant alignment with Aureobasidium namibiae CBS 147.97 hypothetical protein partial mRNA and 46 % with Aureobasidium pullulans JQ624241 and AF470619; Identified sequenced fragment was 2051 bp. and G+C content is 50.5% with molecular mass 63 KDa.

Crude oil spills as a result of natural disasters or extraction and transportation operations are common nowadays. Oil spills have adverse effects on both aquatic and terrestrial ecosystems and pose a threat to human health. This study have been concerned with studying the capability of six fungal species (Curvularia brachyspora, Penicillium chrysogenum, Scopulariopsis brevicaulis, Cladosporium sphaerospermum, Alternaria alternata, and Stemphylium botryosum) and three fungal consortia (FC), FC1 (P. chrysogenum and C. brachyspora), FC2 (S. brevicaulis and S. botryosum), and FC3 (S. brevicaulis, S. botryosum, and C. sphaerospermum), to remediate petroleum hydrocarbons (PHs). Qualitative and quantitative changes in polyaromatic hydrocarbons (PAHs) and saturated hydrocarbons (SH) mixtures and the patterns of PHs degradation have been examined using HPLC and GC. Studying the GC chromatogram of C. sphaerospermum revealed severe degradation of SHs exhibited by this species, and the normal-paraffin and isoparaffin degradation percentage have been valued 97.19% and 98.88%, respectively. A. alternata has shown the highest significant (at P ˂ 0.05) PAH degradation percent reaching 72.07%; followed by P. chrysogenum, 59.51%. HPLC data have revealed that high-molecular-weight PAH percent/total PAHs decreased significantly from 98.94% in control samples to 68.78% in samples treated with A. alternata. FC1 and FC2 consortia have exhibited the highest significant PH deterioration abilities than did the individual isolates, indicating that these fungal consortia exhibited positive synergistic effects. The study supports the critical idea of the potential PAH and SH biodegradation as a more ecologically acceptable alternative to their chemical degradation.

Gum arabic (GA) is a traditional herbal medicine from Acacia Senegal (L.) Willdenow trees, which consist of a complex mixture of polysaccharides and glycoproteins. It is used in daily applications for several diseases and is considered to protect against bacterial infections. The detailed mechanisms behind these observations are still unclear. In this study, we investigated the direct antibacterial activity of GA water and ethanol extracts against Staphylococcus (S.) aureus or Escherichia (E.) coli and the immunomodulating properties of those extracts on granulocytes as a first line of defense against bacteria. Firstly, the direct antimicrobial effect of GA was tested on three different S. aureus strains and two E. coli strains. The growth of bacteria was analyzed in the presence of different GA concentrations over time. GA water as well as ethanol extracts showed a significant growth inhibition in a concentration-dependent manner in the case of S. aureus Newman, S. aureus Rd5, and E. coli 25922, but not in the case of S. aureus USA300 and E. coli K1. Transmission electron microscopic analysis confirmed an antibacterial effect of GA on the bacteria. Secondly, the immunomodulatory effect of GA on the antimicrobial activity of bovine or human blood-derived granulocytes was evaluated. Interestingly, water and ethanol extracts enhanced antimicrobial activity of granulocytes by the induction of intracellular ROS production. In line with these data, GA increased the phagocytosis rate of E. coli. No effect was seen on neutrophil extracellular trap (NET) formation that mediates killing of extracellular bacteria such as S. aureus. In conclusion, we show that GA exhibits a direct antibacterial effect against some S. aureus and E. coli strains. Furthermore, GA boosts the antimicrobial activities of granulocytes and increases intracellular ROS production, which may lead to more phagocytosis and intracellular killing. These data might explain the described putative antimicrobial activity of GA used in traditional medicine.

L-glutaminase is an amidohydrolase that catalyzes L-glutamine to L-glutamic acid and has increased impressive consideration due to its potential anticancer activity. The objective of this work was to isolate different fungi that produce L-glutaminase which has antitumor activities. Three fungal species that were isolated on the selective medium showed L-glutaminase activity. The species were identified as a Penicillium crustosum, Emericella nidulans and Mucor circinelloides. Mucor circinelloides showed potential L-glutaminase production (5.16 U/ml). Different pH, L-glutamine concentrations, temperatures, and incubation periods were carried out. The most enzyme production was shown at pH 8, temperature 30 ºC, with 0.6% of the L- glutamine. L-glutaminase was extracted and purified with gel filtration and ion exchange (DEAE Sephadex A50). The purified enzyme was found to possess a specific activity 1.55 U/mg and was monitored by SDS-PAGE with molecular weight 70 kDa. Purified L-glutaminase exhibited cytotoxic activity against HepG2 and PC3 cell lines with Ic50 value 0.067 mg/ml and 0.079 mg/ml respectively. Caspase 3 colorimetric assay and flow cytometry was done to detect the activation of apoptosis among the enzyme. The enzyme showed positive results on both types of cell lines.

Antimicrobial resistance was declared by the World Health Organization (WHO) in 2014 as the greatest threat for human and veterinary medicine.The development of resistance in E. coli may be due to haphazard use of antibiotics, plasmid-mediated genes,i.e. blaCTX-M, blaSHV, blaOXA. This work was aimed to detect the antibiotic resistant genes in multidrug-resistant extended-spectrum β-lactamase-producing E. coli from samples of children. 90 samples from chidren were collected. blaTEM, blaSHV, blaOXA, blaCTX-M and blaVEB resistance genes in drug-resistant E. coli were investigated. The blaTEM, blaSHV and blaCTX genes were detected in E.coli isolates. blaTEM genes were found to be in all isolates. The blaSHV gene was detected in 14.28% of the isolates. The blaCTX-M gene was detected in 71.42% of the isolates blaOXA and blaVEB genes were not detected in any sample.The resistant genes sequences of the E.coli isolates from urine, blood, skin ande ndotracheal tube were deposited in the DDBJ/EMBL/GenBank nucleotide sequence databases.The resistance genes sequences of the Escherichia isolates are greatest closely associated to those of Escherichia coli.

•The biochemical properties of two proteobacteria laccases were assessed.•Polyextremotolerant qualities of the laccases were identified.•Multiple laccase-encoding genes were observed in laccase-producing strains.•Their implication in biotechnological applications was deliberated.

The aim of this work was performed to develop a milder and cost effective extraction procedure for Beta-glucan (BG) from two types of S. cerevisiae, the returned Baker's yeast “RBY” and yeast remaining after ethanol fermentation “EFY” using different chemical extraction steps. The current results revealed that the carbohydrate percent in yeast biomass was considerably increased by the different extraction steps (from 37.64 to 92.41% and from 41.37 to 93.46% in case of RBY and EFY, respectively). On the other hand, the percent of protein in yeast biomass was decreased by the different extraction steps (from 41.91 to 1.28% and from 36.32 to 1.15% in the two cases, respectively). The extracted dry biomass in the two cases were analysed by Fourier Transform Infrared (FTIR) spectra. This method of β-glucan extraction steps has been shown to produce great yields of β-glucan with maintaining their purity and native structure.

Increasing incidence of multidrug-resistant extended-spectrum beta-lactamase-producing bacteria is a significant health problem that has strongly impacted the treatment of infectious diseases and cancer. Consequently, this work was aimed to investigate the prevalence of drug-resistant E. coli isolated from clinical samples of adults and children. 180 samples (90 from adults and 90 from children) were collected. 130 bacterial isolates (80 from children and 50 from adults) showed to be E. coli. The prevalence of resistance to common antibiotics in adults and children was ampicillin (84% and 69%), cephalothin (72% and 54%), trimethoprim-sulfamethoxazole (62% and 47%), ciprofloxacin (54% and 24%), levofloxacin (54% and 21%), aztreonam (44% and 43%), cefepime (36% and 43%), ceftriaxone (34% and 43%), amoxicillin-clavulanate (34% and 42%), ceftazidime (32% and 43%), gentamicin (16% and 11%), nitrofurantion (4% and 3%), ertapenem (4% and 6%) and piperacillin-tazobactam (2% and 1%), respectively. MDR E. coli (resistance to >= 3 antimicrobial groups) in adults and children were 37 (74%) and 59 (73.75%), respectively. ESBL-producing rate among E. coli isolates was 50% and 44% in adults and children, respectively. Sequences of 16S rRNA genes of the isolates share 99% similarity with that of E. coli strain DX15. The isolation of MDRESBL-producing E. Coli definitely will limit the choices of clinicians to treat their patients. Therefore, there is an urgent requisite for surveillance studies on antimicrobial resistance and incidence of ESBLs among patients to guide the clinical cure.

The increasing occurrence of antibiotic-resistant Staphylococcus (S.) aureus tremendously limits the antibiotic-based treatment options; therefore, an open discussion of alternative treatment strategies is urgently needed. The use of naturally derived materials might become a more promising concept, not only as directly acting antimicrobials, but also for stimulation of the immune system. Costa Rican plant extracts were screened for their ability to enhance the antimicrobial activity of human blood-derived cells against S. aureus infections. We identified three plant extracts which significantly reduced the growth of S. aureus in the presence of human blood without directly acting as antibacterials: Byrsonima crassifolia acetone bark extract, Mandevilla veraguasensis acetone vine extract and Verbesina oerstediana acetone bark extract (VEOEBA). The effect of VEOEBA was studied in more detail, and revealed that VEOEBA increases the antimicrobial activity of neutrophils by enhancing the formation of neutrophil extracellular traps.

PEROXIDASES have numerous important applications in industries and biodegradation of organopollutants , so there is a need to explore more and more sources of enzymes with different characteristics. In the present study, lignin peroxidases (LiPs) producing by fungus Humicola grisea were extracted and purified using gel filtration and ion exchange chromatography. Protein and hems protein were eluted in different peaks about 6 of these peaks had LiP activity and this is an indication that LiP in H. grisea was isoenzymes. Four of these isoenzymes were characterized regarding their molecular mass, optimum temperature and optimum pH for activity, substrate concentration and effect of some metal ions. The four purified isoenzymes have maximum activities at different temperatures, where optimum temperature for H2 and H4isoenzyme were at 60 degrees C, and the optimum temperatures for HI and H3 were at 40 degrees C and 30 degrees C, respectively. The maximum activities of the four isoenzymes were within the acidic range. The different LiP isoenzymes showed different Kin and Vmax values representing the varied affinity of the four isoenzymes using veratryl alcohol as a substrate. Km values for HI , H2, H3 and H4 were 5.26, 5.0, 1.3 and 1.4, respectively. While Vmax values for the same isoenzymes were 1.25, 0.8, 0.87 and 1.1, respectively.

* Corresponding Author: monierabdelghani@yahoo.com ABSTRACT The morphological variations between selected five Galium species from Libya, and the role of soil factors that affect their distribution in their natural habitats were investigated. Thirty-seven macromorphological characters (11quantitative, 26 qualitative) representing vegetative parts were subjected to numericaltaxonomic analysis. Five branches and clusters were distinguished; each was linked to a specific species. Representatives of these groups were clustered together according to characters with high factor loading in the Principal Components Analysis (PCA). The results showed congruence between the UPGMA clustering and PCA in suggesting five species groups. Seventeen soil factors were used in this assessment of soil factors responsible for the distribution of the 5 species of Galium. Calcium and chloride ions content exhibited the most significant difference (p=0.05) among the five species groups, while the other examined soil variables showed no significant differences. The relationship between the examined soil variables and the studied populations of Galium species was assessed by Principal Components Analysis (PCA). Each of the studied species of Galium was affected by one or more of the examined soil parameters.

THE AIM of the current investigation was to study biodiesel production by Fusarium oxysporum AUMC 3224 cultured on two types of agricultural wastes , the first one called el-ghasheem and the second was wheat straw. In the case of el-ghasheem medium, the maximum lipid production was attained with 125 gl(-1) ghasheem concentration and 0.1 gl(-1) NaNO3 for 7 days, at pH 5.5 and temperature 28 degrees C. Maximum lipid yield was 1.558 +/- 0.003 gl(-1) lipid weight and 32.44 +/- 0.035% lipid percentage. In the case of wheat straw, the maximum lipid production was achieved with 5 g wheat straw, 50 ml of 2 gl(-1) NaNO3, for 14 days, at 28 degrees C. Maximum lipid yield was 0.45 +/- 0.005 g/L lipid weight and 28.81 +/- 0.305 % lipid percentage. The profile of extracted lipids from F. oxysporum was studied using Fourier transform infrared spectrometer (FTIR) indicating the presence of triglycerides and after transesterification of lipids showed the presence of fatty acid methyl esters. The fatty acids profiles were also determined by gas chromatography (GC) coupled with flame ionization detector and GC-mass spectroscopy. Data revealed the presence of significant amounts of palmitic, oleic, stearic, linoleic and other methyl esters.The results showed that lipids from F.oxysporum was a potential alternative resource for biodiesel production.

The ability of Bassia scoparia (L) A.J. Scott to remediate petroleum-contaminated arid land sandy soil was studied with natural and sterilized soils, and with supplemental nutrients and water. The species showed good tolerance of petroleum hydrocarbons (PHs) in soils reaching 2-3% (oil:soil by mass) pollution levels. After five months of phytoremediation, the average degradation rate of petroleum hydrocarbons ranged between 31.2 +/- 1.15-57.7 +/- 1.29% for natural soil and 28.7 +/- 1.04-51.1 +/- 1.53% for pre-sterilized soil. The highest breakdown of PHs for both saturated and poly-aromatic fractions was achieved when plants were present. Changes in saturated and aromatic fractions were monitored and measured using gas chromatography and high performance liquid chromatography. Moderate concentrations of PHs activated specialized oil-degrading microorganisms which in turn promoted the efficiency of phytoremediation. Polluted soils planted with B. scoparia also showed a significant reduction in sulfur levels. The potential demonstrated for remediation of petroleum hydrocarbons and sulfur by B. scoparia suggests it may be a useful tool for remediation of arid land soils contaminated with crude oil. (C) 2014 Elsevier Ltd. All rights reserved.