Paecilomyces variotii xylanase was, produced in stirred tank bioreactor with yield of 760 U/mL and purified using 70% ammonium sulfate precipitation and ultra-filtration causing 3.29-fold purification with 34.47% activity recovery. The enzyme purity was analyzed on sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) confirming its monomeric nature as single band at 32 KDa. Zymography showed xylan hydrolysis activity at the same band. The purified enzyme had optimum activity at 60 °C and pH 5.0. The pH stability range was 5–9 and the temperature stability was up 70 °C. Fe2+and Fe3+ exhibited inhibition of xylanase enzyme while Cu2+, Ca2+, Mg2+ and Mn2+ stimulated its activity. Mercaptoethanol stimulated its activity; however, Na2-EDTA and SDS inhibited its activity. The purified xylanase could hydrolyze beechwood xylan but not carboxymethyl cellulose (CMC), avicel or soluble starch. Paecilomyces variotii xylanase Km and Vmax for beechwood were determined to be 3.33 mg/mL and 5555 U/mg, respectively. The produced xylanase enzyme applied on beech xylan resulted in different types of XOS. The antioxidant activity of xylo-oligosaccharides increased from 15.22 to 70.57% when the extract concentration was increased from 0.1 to 1.5 mg/mL. The enzyme characteristics and kinetic parameters indicated its high efficiency in the hydrolysis of xylan and its potential effectiveness in lignocellulosic hydrolysis and other industrial application. It also suggests the potential of xylanase enzyme for production of XOS from biomass which are useful in food and pharmaceutical industries.
The risk of resistance development and adverse effects on human health and the environment has increased in the last decade. Furthermore, many antifungal agents fail to inhibit the pathogenesis of azole-resistant Aspergillus flavus. In this report, we isolated and identified azole-resistant A. flavus isolates from two sources of maize (white and yellow maize). The susceptibilities of Aspergillus flavus isolates were investigated by conventional antifungals such as Terbinfine, Fluconazole, Ketoconazole, Voricazole, Amphotericin, and Nystatin. Then zinc oxide nanoparticles associated with Chlorella vulgaris, which are synthesized by using the precipitation method, were examined against isolated fungi. The results showed that twelve species of white corn were isolated out of fifty isolates, while the number of isolates from the yellow corn source was only four. Interestingly, the following antifungals have an impact effect against azole-resistant A. flavus isolates: the inhibition zones of ketoconazole, voricazole, and terbinafine were 40 mm, 20 mm, and 12 mm, respectively, while the remaining antifungal agents have no effect. Similarly, the inhibition zones of the following antifungal agents were as follows: 41 mm for Terbinfine, 13 mm for Voricazole, and 11 mm for Ketoconazole against Aspergillus flavus that was isolated from yellow corn. The physiochemical characterization of zinc oxide nanoparticles provides evidence that ZnO-NPs associate with Chlorella vulgaris and have been fabricated by the precipitation method with a diameter of 25 nm. The zinc oxide nanoparticle was then used to isolate azole-resistant A. flavus, and the results show that ZnO-NPs have an effect on azole-resistant A. flavus isolation. The inhibition zone of zinc oxide nanoparticles against A. flavus (that was isolated from white corn) was 50 mm with an MIC of 50 mg/mL, while the inhibition zone of zinc oxide nanoparticles against Azole-resistant A. flavus isolated from yellow corn was 14 nm with an MIC of 25 mg/mL, which indicated that zinc oxide nanoparticles gave a better result against Azole-resistant A. flavus isolated from maize.
Aspergillus sydowii is a mesophilic soil saprobe that is a food contaminant as well as a human pathogen in immune-compromised patients. The biological fabrication of silica and silver nanoparticles provides advancements over the chemical approach, as it is eco-friendly and cost-effective. In the present study, Aspergillus sydowii isolates were collected from the soil fields of six different sites in the western area of Saudi Arabia and then identified using the PCR technique following sequencing analysis by BLAST and phylogenetic analysis. Then, applied silica and silver nanoparticles were synthesized by biological methods, using Aspergillus niger as a reducer. Silver and silica nanoparticles were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The antifungal activity of silver and silica nanoparticles against Aspergillus sydowii isolates was evaluated using the disc diffusion method and the minimum inhibitory concentration (MIC). The physiochemical results emphasized the fabrication of silver and silica nanoparticles in spherical shapes with a diameter in the range of 15 and 40 nm, respectively, without any aggregation. MIC of Ag-NPs and Si-NPs against Aspergillus sydowii isolates were 31.25 and 62.5 µg/mL, respectively. Finally, the aim of the study is the use of silver as well as silica nanoparticles as antifungal agents against Aspergillus sydowii.
Harmful algal blooms (HABs) and the high biomass associated with them have afflicted marine desalination plants along coastal regions around the world. Few studies of HABs have been conducted in the Red Sea, where desalination plants along the Saudi Arabian Red Sea coast provide drinking water for millions of people. This study was conducted along the Saudi Arabian Red Sea coast from 2014 to 2015 to assess the potential for using Moderate Resolution Imaging Spectroradiometer (MODIS) remote sensing of chlorophyll a (Chl a) or fluorescence line height (FLH) to identify risks for biofouling at these desalination plants. Ship-based surveys of phytoplankton were conducted along the Saudi Arabian coastline offshore of desalination plants at Jeddah, Al Shoaibah and Al Qunfudhuh to assess the density of phytoplankton populations and identify any potential HAB species. Ship-based surveys showed low to moderate concentrations of phytoplankton, averaging from 1800–10,000 cells L−1 at Jeddah, 2000–11,000 cells L−1 at Al Shoaibah and 1000–20,500 cells L−1 at Al Qunfudhuh. Sixteen different species of potentially toxigenic HABs were identified through these surveys. There was a good relationship between ship-based total phytoplankton counts and monthly averaged coastal MODIS Chl a (R2 = 0.49, root mean square error (RMSE) = 0.27 mg m−3) or FLH (R2 = 0.47, RMSE = 0.04 mW m−2 µm−1 sr−1) values. Monthly average near shore Chl a concentrations obtained using MODIS satellite imagery were much higher in the Red Sea coastal areas at Al Qunfudhuh (maximum of about 1.3 mg m−3) than at Jeddah or Al Shoaibah (maximum of about 0.4 and 0.5 mg m−3, respectively). Chlorophyll a concentrations were generally highest from the months of December to March, producing higher risks of biofouling desalination plants than in other months. Concentrations decreased significantly, on average, from April to September. Long-term (2005–2016) monthly averaged MODIS Chl a values were used to delineate four statistically distinct zones of differing HAB biomass across the entire Red Sea. Sinusoidal functions representing monthly variability were fit to satellite Chl a values in each zone (RMSE values from 0.691 to 0.07 mg m−3, from Zone 1 to 4). December to January mean values and annual amplitudes for Chl a in these four sinusoidal functions decreased from Zones 1–4. In general, the greatest risk of HABs to desalination occurs during winter months in Zone 1 (Southern Red Sea), while HAB risks to desalination plants in winter months are low to moderate in Zone 2 (South Central Red Sea), and negligible in Zones 3 (North Central) and 4 (Northern).
In this work, sequential optimization strategy, based on statistical designs, was employed to enhance the degradation of PHB by Trichoderma asperellum through the measurement of PHB depolymerase specific activity. For PHB degradation screening, two-level Plackett-Burman design was used. Result of study revealed that fermentation medium composition significantly influencing the PHB depolymerase specific activity. Further, it was reported that time of incubation, levels of PHB and concentration of glucose were found to be the major factors influencing the enzyme specific activity. A second threelevel Box-Behnken design was applied to acquire the best process conditions for PHB depolymerase specific activity. In this study, the initial basal medium showed a PHB depolymerase specific activity of 40 ng / l / hr / g dry wt and through the two successive statistical design, the final level of PHB depolymerase specific activity was 3230 ng / l / hr / g drywt, this means a nearly of 80 fold increase.
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