Xylitol bioproduction by Candida tropicalis: effects of glucose/xylose ratio and pH on fermentation and gene expression.

Abstract

Xylitol is a highly demanded polyol in the food, pharmaceutical, and chemical industries. However, its current production methods are considered energy-intensive, require the use of hazardous chemical catalysts, and depend on complex and costly equipment. The biotechnological route of xylitol production is proposed as a sustainable alternative, but it still requires process improvements, such as enhanced fermentation capabilities, to be economically competitive. This study examined Candida tropicalis yeast to improve xylose-to-xylitol conversion via glucose: xylose ratio and pH modulation. Key parameters evaluated included xylose consumption rate (r_S), xylose-to-xylitol yield (Y_P/S), and xylitol volumetric productivity (Q_P). Conditions with 50 g/L xylose at pH 3.5 exhibited superior xylitol production: 29.81 g/L, Q_P of 0.52 g/L/h, and Y_P/S of 0.54 g/g at 48 h. The statistical model demonstrated that the maximum Y_P/S and Q_P values have not yet been achieved. This could present an opportunity to be explored through yeast genetic engineering approaches. Additionally, the quantitative expression of the xylose transporter genes (XUT1 and STL2) and the xylose reductase gene (XYL1), previously identified in C. tropicalis, was evaluated under all tested conditions. Upregulation of the XUT1 was correlated with higher xylose concentrations, while STL2 was favored at lower xylose concentrations. The expression of XYL1 showed upregulation over time with higher xylose ratios. The high transcription levels and expression profile suggest that Xut1p-mediated xylose transport occurs through a proton symport mechanism. The results indicate that the pH factor indirectly influences XUT1 gene transcription, possibly as a compensatory response to the reduced transporter efficiency under high pH conditions. The present work underscores the influence of glucose ratios and pH in xylitol production, as well as the gene expression of xylose transporters and the key enzyme xylose reductase. Leveraging these insights can significantly enhance xylitol production from hemicellulosic hydrolysates through biotechnological pathways.