Application of Ligninolytic Enzymes in the Production of Biofuels from Cotton Wastes

The application of ligninolytic fungi and enzymes is an option to overcome the issues related with the production of biofuels using cotton wastes. In this dissertation, the ligninolytic fungus and enzymes were evaluated as pretreatment for the biochemical conversion of Cotton Gin Trash (CGT) in ethanol and as a treatment for the transformation of cotton wastes biochar in other substances. In biochemical conversion, seven combinations of three pretreatments (ultrasonication, liquid hot water and ligninolytic enzymes) were evaluated on CGT. The best results were achieved by the sequential combination of ultrasonication, hot water, and ligninolytic enzymes with an improvement of 10% in ethanol yield. To improve these results, alkaline-ultrasonication was evaluated. Additionally, Fourier Transform Infrared (FT-IR) and principal component analysis (PCA) were employed as fast methodology to identify structural differences in the biomass. The combination of ultrasonication-alkali hydrolysis, hot liquid water, and ligninolytic enzymes using 15% of NaOH improved 35% ethanol yield compared with the original treatment. Additionally, FT-IR and PCA identified modifications in the biomass structure after different types of pretreatments and conditions. In thermal conversion, this study evaluated the biodepolymerization of cotton wastes biochar using chemical and biological treatments. The chemical depolymerization evaluated three chemical agents (KMnO4, H2SO4, and NaOH), with three concentrations and two environmental conditions. The sulfuric acid treatments performed the largest transformations of the biochar solid phase; whereas, the KMnO4 treatments achieved the largest depolymerizations. The compounds released into the liquid phase were correlated with fulvic and humic acids and silicon compounds. The biological depolymerization utilized four ligninolytic fungi Phanerochaete chrysosporium, Ceriporiopsis subvermispora, Postia placenta, and Bjerkandera adusta. The greatest depolymerization was obtained by C. subvermispora. The depolymerization kinetics of C. subvermispora evidenced the production of laccase and manganese peroxidase and a correlation between depolymerization and production of ligninolytic enzymes. The modifications obtained in the liquid and solid phases showed the production of humic and fulvic acids from the cultures with C. subvermispora. The results of this research are the initial steps for the development of new processes using the ligninolytic fungus and their enzymes for the production of biofuels from cotton wastes

Propagación in vitro de plantas adultas de Vaccinium meridionale (Ericaceae) In vitro propagation of mature plants of Vaccinium meridionale (Ericaceae)

Se desarrolló un procedimiento de micropropagación de plantas adultas de Vaccinium meridionale utilizando como explantes primarios ápices caulinares. Durante la fase de establecimiento in vitro de explantes se estudió el efecto de los medios MS/3, WPM, AND y el propuesto por Kyte para Blueberry, suplementados con 2-iP más AIA ó BA más AIA. Durante la proliferación de microtallos se evaluó el efecto del medio MS/3 líquido, sólido y en doble fase (una fase líquida sobre una fase solidificada con agar), suplementados con 2-iP más AIA. El enraizamiento in vitro y ex vitro de microtallos y macollas se indujo utilizando auxinas y/o carbón activado y para el desarrollo de raíces se utilizó un sustrato enriquecido con materia orgánica. El endurecimiento de plántulas se realizó de manera simultánea con el proceso de desarrollo radical. Después de la fase de establecimiento, la media más alta, 4.5, yemas axilares desarrolladas por explante viable, se cuantificó en MS/3 suplementado con 59.05 µM de 2-iP más 17.13 µM de AIA. Durante la fase de proliferación de microtallos la media más alta, 7.25, se cuantificó en MS/3 en doble fase. Después de 60 días de endurecimiento el 88-100% de los microtallos enraizaron y reactivaron su crecimiento.<br>Using stem apex as primary explants, a micropropagation protocol of Vaccinium meridionale was established. During establishment phase the effect of the MS/3, WPM, AND and Kyte media, supplemented with 2-iP plus IAA or BA plus IAA was studied. During microshoot proliferation the effect of MS/3 liquid, solid and double phase (the liquid phase in a solidified phase with agar) supplemented with 2-iP plus IAA was evaluated. In vitro and ex vitro rooting of microshoots and microshoots was accomplished using auxines and/or activated charcoal; for root development a substratum with abundant organic matter was utilized. Plantlet hardening was achived simultaneously with the radical development process. After establishment phase, the highest quantity of axillary buds/explant was quantified in cultures performed in MS/3 supplemented with 2-iP, 59.05 µM plus IAA 17.13 µM. During the microshoot proliferation phase the highest average production was obtained in double phase MS/3. After 60 days of hardening 88-100% of rooted microshoots was obtained; these plantlets showed growth reactivation