RESUMEN
Inhibitors released by the breakdown of plant cell walls prevent efficient conversion of sugar into ethanol. The aim of this study was to develop a fast and reliable inhibitor sensitivity assay for ethanologenic yeast strains. The assay comprised bespoke 96-well plates containing inhibitors in isolation or combination in a format that was compatible with the Phenotypic Microarray Omnilog reader (Biolog, hayward, CA, USA). A redox reporter within the assay permits analysis of inhibitor sensitivity in aerobic and/or anaerobic conditions. Results from the assay were verified using growth on spot plates and tolerance assays in which maintenance of viability was assessed. The assay allows for individual and synergistic effects of inhibitors to be determined. It was observed that the presence of both acetic and formic acid significantly inhibited the yeast strains assessed, although this impact could be partially mitigated by buffering to neutral pH. Scheffersomyces stipitis, Candida spp., and Pichia guilliermondii demonstrated increased sensitivity to short chain weak acids at concentrations typically present in lignocellulosic hydrolysates. S. cerevisiae exhibited robustness to short chain weak acids at these concentrations. However, S. stipitis, Candida spp., and P. guilliermondii displayed increased tolerance to HMF when compared to that observed for S. cerevisiae. The results demonstrate that the phenotypic microarray assay developed in the current study is a valuable tool that can be used to identify yeast strains with desirable resistance to inhibitory compounds found in lignocellulosic hydrolysates.
Asunto(s)
Etanol/metabolismo , Lignina/metabolismo , Pruebas de Toxicidad/métodos , Levaduras/metabolismo , Ácido Acético/toxicidad , Alimentación Animal , Candida/efectos de los fármacos , Candida/metabolismo , Fermentación , Formiatos/toxicidad , Análisis por Micromatrices , Fenotipo , Pichia/metabolismo , Saccharomyces cerevisiae/efectos de los fármacos , Saccharomyces cerevisiae/metabolismo , Saccharomycetales/efectos de los fármacos , Saccharomycetales/metabolismo , Xilosa/metabolismo , Levaduras/efectos de los fármacos , Levaduras/crecimiento & desarrolloRESUMEN
A multiplex technique is described that is binary in nature and that can be used in the observation of a dispersed spectrum in any wavelength region for which a detector exists. The subsequent data processing involves the minimum possible number of operations and can easily be carried out in real time. The number of observations that must be made is also a minimum, equal to the number of spectral elements. Methods of generating the coding and decoding patterns, derived from the simple mathematical formulation of the multiplexing technique, are described.