Application of artificial neural networks for enhancing Aspergillus flavipes lipase synthesis for green biodiesel production.

Biodiesel is a sustainable, and renewable alternative to fossil fuels that can be produced from various biological sources with the aid of lipases. This study developed a simple and novel fungal system for lipase biosynthesis to be used for catalyzing the oily residuals into biodiesel, employing the artificial neural network (ANN), and semi-solid-state fermentation (SSSF). Nigella sativa was selected among agro-industrial oily residuals as a substrate for lipase biosynthesis by Aspergillus flavipes MH47297. The effect of cultural humidity (X1), the surfactant; Brij 35 (X2), and inoculum density (X3) on lipase biosynthesis were researched based on the matrix of Box-Behnken design (BBD). The ANN together with a new fungal candidate and SSSF were then applied for the first time to model the biosynthesis process of lipase. The optimum predicted cultural conditions varied according to the model. The optimum predicted conditions were estimated separately by BBD (X1 = 5.8 ml water/g, X2 = 46.6 µl/g, and X3 = 62156610 spore/g) and ANN (X1 = 5.4 ml water/g, X2 = 54.2 µl/g, and X3 = 100000000 spore/g) models. Based on the modeling process, the response of lipase was calculated to be 214.95 (BBD) and 217.72 U (ANN), which revealed high consistency with the experimental lipase yield (209.13 ± 3.27 U for BBD, and 218 ± 2.01 U for ANN). Despite both models showing high accuracy, ANN was more accurate and surpassed the BBD model. Gas chromatography analysis showed that lipase successfully converted corn oil to biodiesel (29.5 mg/l).

Production, characterization and bio-emulsifying application of exopolysaccharides from Rhodotorula mucilaginosa YMM19.

Microbial exopolysaccharides (EPS) are high molecular weight polymers having different sugar residues. EPS have potential applications in different fields, such as medicine, food and environment. Therefore, there is a growing interest in production, characterization and application of EPS from different microorganisms. The present study designed to investigate the production and characterization of EPS from Rhodotorula mucilaginosa YMM19 isolated from Morus nigra L. fruits as well as to examine their potential emulsifying properties. Effect of NaCl concentration, incubation period and pH on the production of EPS was studied. The maximum EPS production by yeast was achieved at 10% NaCl (9741.84 mg/l). The best incubation time for production of EPS was 5 days. Production of EPS decreased under neutral condition and increased at acidic and alkaline condition. The structural feature of EPS was examined by FT-IR and NMR spectral analysis and confirmed the presence of glucose, glucopyranose and galactose. The isolated EPS showed higher emulsification capacity with emulsification activity of 71% and emulsifying index of 60%. The EPS gave strong emulsification for farnesol and was more effective than sodium dodecyl sulphate, a reference emulsifier, in enhancing the herbicidal activity of farnesol against Melilotus indicus under greenhouse condition. The results suggest that the EPS produced by YMM19 strain has a potential to be used as emulsifying agent in pesticide formulations.

Applications of Fusarium solani YMM20 in bioremediation of heavy metals via enhancing extracellular green synthesis of nanoparticles.

A novel technique for removing metals from water, by converting them into nanoparticles using the culture filtrate of Fusarium solani YMM20, followed by their removal by centrifugation was developed in this study. It is a promising, simple, and eco-friendly process for the bioremediation of wastewater. This method is built on the reduction of metal ions by cell-free fungal filtrate protein(s). The chitosan and chitosan nanoparticles were used as adsorbents for the removal of Cu2+ , Pb2+ , Co2+ , Cd2+ , Ni2+ , and Fe2+ , separately. In addition, fungal filtrate and chitosan nanoparticles loaded with fungal filtrate were used for the biosynthesis of metal nanoparticles of Cu2+ , Pb2+ , Co2+ , Cd2+ , Ni2+ , and Fe2+ , separately for their removal. The fungal filtrate approach has the highest metal removal (%) for most metals. Among the six metal ions, the fungal filtrate, chitosan nanoparticles loaded with fungal filtrate, chitosan, and chitosan nanoparticles showed the highest removal (%) for Pb2+ and the lowest removal (%) for Co2+ . Techniques such as transmission electron microscopy, scanning electron microscopy, and Zeta potential measurements were used to characterize the obtained nanoparticles. The average size of the spherical nanoparticles observed by transmission electron microscopy was from 18.938 ± 2.269 nm to 60.175 ± 4.973 nm.

Optimization of methylene blue degradation by Aspergillus terreus YESM 3 using response surface methodology.

Synthetic dyes released from many industries cause pollution problems in aquatic environments affecting public health. The present study aimed to explore the potentiality of Aspergillus terreus YESM 3 (accession number LM653117) for colour removal of three different dyes: methylene blue (MB), malachite green (MG) and safranin (S). Results showed that the tolerance index of the studied fungus against tested dyes decreased in the order: methylene blue, safranin and malachite green. Removal of methylene blue colour was improved by using Box-Behnken design. Optimum condition for methylene blue biodegradation in Czapek Dox broth was achieved at pH 6, of 31.41 mg/L dye concentration and an inoculum of 5.7778 × 104 (conidia/mL) with biodegradation of 89.41%. Thus, a novel and eco-friendly system for the biodegradation of dyes using Box-Behnken design has been efficiently developed. Accordingly, A. terreus YESM 3 can be professionally used for bioremediation of methylene blue dye in wastewater and removal of environmental pollution.

Isolation and Identification of Plant Growth-Promoting Bacteria Highly Effective in Suppressing Root Rot in Fava Beans.

In this study, 18 plant growth-promoting bacterial (PGPB) strains were isolated from the rhizosphere of both Red silk-cotton tree (Bombax malabaricum) and Chinese banyan (Ficus retusa). Culture morphology was observed and genotypic characterization was accomplished by sequencing partial 16S rRNA gene. Plant growth promotion traits and antagonistic activities of the strains against phytopathogenic fungi were evaluated. Among all strains, Bacillus thuringiensis (MN419208) exert the highest indole acetic acid (38 µg/ml), produced exopolysaccharides (587.2 µg/ml), and fixed nitrogen which in turn increased both fresh and dry weights of bean plants by 41.5% and 18.8%, respectively. In another greenhouse experiment studying the antifungal activities of seven strains and their co-culture against Rhizoctonia solani (LN735538), B. sonorensis MN419205, B. wiedmannii MN419207, B. subtilis MN419218, and the mixture of (MN419207) and (MN419208) reduced total damping off from 81.7% in control to 30%, 35%, 35%, and 38.3%, respectively, and reduced disease severity index from 33.3% to 20.5%, 22.5%, 14.2%, and 19.3% as well. Our data indicate that these strains are effective in promoting plant growth and in inhibiting R. solani infection nonetheless field experiments are needed to examine their effectiveness as a viable alternative to chemical fertilizers and conventional pesticides.

Biodegradation of imidacloprid in liquid media by an isolated wastewater fungus Aspergillus terreus YESM3.

In the present study, a new fungal strain capable of imidacloprid degradation was isolated from agricultural wastewater drain. The fungal strain of YESM3 was identified as Aspergillus terreus based on ITS1-5.8S rDNA-ITS2 gene sequence by PCR amplification of a 500 bp sequence. Screening of A. terreus YESM3 to the insecticide imidacloprid tolerance was achieved by growing fungus in Czapek Dox agar for 6 days at 28°C. High values (1.13 and 0.94 cm cm-1) of tolerance index (TI) were recorded at 25 and 50 mg L-1 of imidacloprid, respectively in the presence and absence of sucrose. However, at 400 mg L-1 the fungus did not grow. Effects of the imidacloprid concentration, pH, and inoculum size on the biodegradation percentage were tested using Box-Behnken statistical design and the biodegradation was monitored by HPLC analysis at different time intervals. Box-Behnken results indicated that optimal conditions for biodegradation were at pH 4 and two fungal discs (10 mm diameter) in the presence of 61.2 mg L-1 of imidacloprid. A. terreus YESM3 strain was capable of degrading 85% of imidacloprid 25 mg L-1 in Czapek Dox broth medium at pH 4 and 28°C for 6 days under static conditions. In addition, after 20 days of inoculation, biodegradation recorded 96.23% of 25 mg L-1 imidacloprid. Degradation kinetics showed that the imidacloprid followed the first order kinetics with half-life (t50) of 1.532 day. Intermediate product identified as 6-chloronicotinic acid (6CNA) as one of the major metabolites during degradation of imidacloprid by using HPLC. Thus, A. terreus YESM3 showed a potential to reduce pollution by pesticides and toxicity in the effected environment. However, further studies should be conducted to understand the biodegradation mechanism of this pesticide in liquid media.