Lignocellulosic wastes, rice straw, sugarcane bagasse, rice bran and sawdust, and pure commercial carboxymethyl cellulose (CMC) and xylan were used as substrates to cultivate cellulolytic fungus, Aspergillus flavus KUB2, in submerged fermentation at 30°C. Of all the substrates, sugarcane bagasse was a good source for the production of cellulolytic and also hemicellulolytic enzymes. The maximum activities of endoglucanase (CMCase), total cellulase (FPase) and xylanase using sugarcane bagasse as substrate were 8%, 75% and 165%, respectively, higher than those of the commercial substrates. The time course determination of enzyme production revealed that the highest CMCase (1.27 U/ml), FPase (0.72 U/ml) and xylanase (376.81 U/ml) activities were observed at 14 days of fermentation. Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) analyses confirmed the efficient structural alteration of sugarcane bagasse caused by enzymatic actions during A. flavus KUB2 cultivation. Based on the results of the hydrolytic enzyme activities, FTIR and SEM, A. flavus KUB2 is suggested as an efficient hydrolytic enzymes producer and an effective lignocellulose degrader, while sugarcane bagasse can be applied as a low-cost carbon source for the economical production of lignocellulose hydrolytic enzymes by A. flavus KUB2.
Ethanol tolerance is one of the important characteristics of ethanol-producing yeast. This study focused on the improvement of ethanol tolerance of Saccharomyces cerevisiae NR1 for enhancing ethanol production by random UV-C mutagenesis. One ethanol-tolerant mutant, UVNR56, displayed a significantly improved ethanol tolerance in the presence of 15% (v/v) ethanol and showed a considerably higher viability during ethanol fermentation from sugarcane molasses and sugarcane molasses with initial ethanol supplementation. A maximum ethanol concentration produced from molasses medium at 37°C by UVNR56 was 10.3% (v/v), productivity of 1.7 g/l/h and a theoretical yield of 98.7%, while the corresponding values for the wild-type were 8.6% (v/v), 1.4 g/l/h and 83.3%, respectively. In addition, during molasses fermentation under initial supplementation of 5% (v/v) ethanol, the maximum ethanol concentration and productivity of UVNR56 was 25.7% and 42.9% higher than the wild-type, respectively.
Effective conversion of xylose into ethanol is important for lignocellulosic ethanol production. In the present study, UV-C mutagenesis was used to improve the efficiency of xylose fermentation. The mutated Scheffersomyces shehatae strain TTC79 fermented glucose as efficiently and xylose more efficiently, producing a higher ethanol concentration than the wild-type. A maximum ethanol concentration of 29.04 g/L was produced from 71.31 g/L xylose, which was 58.95 % higher than that of the wild-type. This mutant also displayed significantly improved hydrolysate inhibitors tolerance and increased ethanol production from non-detoxified lignocellulosic hydrolysates. The ethanol yield, productivity and theoretical yield by TTC79 from sugarcane bagasse hydrolysate were 0.46 g/g, 0.20 g/L/h and 90.61 %, respectively, while the corresponding values for the wild-type were 0.20 g/g, 0.04 g/L/h and 39.20 %, respectively. These results demonstrate that S. shehatae TTC79 is a useful non-recombinant strain, combining efficient xylose consumption and high inhibitor tolerance, with potential for application in ethanol production from lignocellulose hydrolysates.
Silver/silver chloride nanoparticles (Ag/AgCl NPs) are an alternative approach to control the larvae of Aedes aegypti, a vector of mosquito-borne diseases. However, the molecular mechanisms of Ag/AgCl NPs to A. aegypti have not been reported. In this work, Ag/AgCl NPs were synthesized using supernatant, mixed toxins from Bacillus thuringiensis subsp. israelensis (Bti), and heterologously expressed Cry4Aa and Cry4Ba toxins. The images from scanning electron microscopy revealed that the Ag/AgCl NPs were spherical in shape with a size range of 25–100 nm. The larvicidal activity against A. aegypti larvae revealed that the Ag/AgCl NPs synthesized using the supernatant of Bti exhibited higher toxicity (LC50 = 0.133 μg/mL) than the Ag/AgCl NPs synthesized using insecticidal proteins (LC50 = 0.148–0.217 μg/mL). The proteomic response to Ag/AgCl NPs synthesized using the supernatant of Bti in A. aegypti larvae was compared to the ddH2O-treated control. Two-dimensional gel electrophoresis analysis revealed 110 differentially expressed proteins, of which 15 were selected for identification using mass spectrometry. Six upregulated proteins (myosin I heavy chain, heat shock protein 70, the F0F1-type ATP synthase beta subunit, methyltransferase, protein kinase, and condensin complex subunit 3) that responded to Ag/AgCl NP treatment in A. aegypti were reported for NP treatments in different organisms. These results suggested that possible mechanisms of action of Ag/AgCl NPs on A. aegypti larvae are: mitochondrial dysfunction, DNA and protein damage, inhibition of cell proliferation, and cell apoptosis. The findings from this work provide greater insight into the action of green synthesized Ag/AgCl NPs on the control of A. aegypti larvae.