Metabolic and Transcriptional Analysis of Recombinant Saccharomyces Cerevisiae for Xylose Fermentation: A Feasible and Efficient Approach.

Lignocellulose is an abundant xylose-containing biomass found in agricultural wastes, and has arisen as a suitable alternative to fossil fuels for the production of bioethanol. Although Saccharomyces cerevisiae has been thoroughly used for the production of bioethanol, its potential to utilize lignocellulose remains poorly understood. In this work, xylose-metabolic genes of Pichia stipitis and Candida tropicalis, under the control of different promoters, were introduced into S. cerevisiae. RNA-seq analysis was use to examine the response of S. cerevisiae metabolism to the introduction of xylose-metabolic genes. The use of the PGK1 promoter to drive xylitol dehydrogenase (XDH) expression, instead of the TEF1 promoter, improved xylose utilization in "XR-pXDH" strain by overexpressing xylose reductase (XR) and XDH form C. tropicalis, enhancing the production of xylitol (13.66 ± 0.54 g/L after 6 days fermentation). Overexpression of xylulokinase and XR/XDH from P. stipitis remarkably decreased xylitol accumulation (1.13 ± 0.06 g/L and 0.89 ± 0.04 g/L xylitol, respectively) and increased ethanol production (196.14 % and 148.50 % increases during the xylose utilization stage, respectively), in comparison with the results of XR-pXDH. This result may be produced due to the enhanced xylose transport, Embden-Meyerhof and pentose phosphate pathways, as well as alleviated oxidative stress. The low xylose consumption rate in these recombinant as well as alleviated strains comparing with P. stipitis and C. tropicalis may be explained by the insufficient supplementation of NADPH and NAD