Contribution of YPRO15C Overexpression to the Resistance of Saccharomyces cerevisiae BY4742 Strain to Furfural Inhibitor

Abstract

Lignocellulosic biomass is still considered a feasible source of bioethanol production. Saccharomyces cerevisiae can adapt to detoxify lignocellulose-derived inhibitors, including furfural. Tolerance of strain performance has been measured by the extent of the lag phase for cell proliferation following the furfural inhibitor challenge. The purpose of this work was to obtain a tolerant yeast strain against furfural through overexpression of YPR015C using the in vivo homologous recombination method. The physiological observation of the overexpressing yeast strain showed that it was more resistant to furfural than its parental strain. Fluorescence microscopy revealed improved enzyme reductase activity and accumulation of oxygen reactive species due to the harmful effects of furfural inhibitor in contrast to its parental strain. Comparative transcriptomic analysis revealed 79 genes potentially involved in amino acid biosynthesis, oxidative stress, cell wall response, heat shock protein, and mitochondrial-associated protein for the YPR015C overexpressing strain associated with stress responses to furfural at the late stage of lag phase growth. Both up- and down-regulated genes involved in diversified functional categories were accountable for tolerance in yeast to survive and adapt to the furfural stress in a time course study during the lag phase growth. This study enlarges our perceptions comprehensively about the physiological and molecular mechanisms implicated in the YPR015C overexpressing strain’s tolerance under furfural stress. Construction illustration of the recombinant plasmid. a) pUG6-TEF1p-YPR015C, b) integration diagram of the recombinant plasmid pUG6-TEF1p-YPR into the chromosomal DNA of Saccharomyces cerevisiae.

An Evolved Strain of the Oleaginous Yeast Rhodotorula toruloides, Multi-Tolerant to the Major Inhibitors Present in Lignocellulosic Hydrolysates, Exhibits an Altered Cell Envelope

“…There are examples of successful alterations of the physicochemical properties of the yeast cell wall either by ALE or by genetic engineering impacting yeast tolerance to stress(es) [33]. The important role played by the cell wall in yeast adaptation and tolerance to different stresses of biotechnological interest clearly emerges from a recent review article [51]. Results here described are in line with those reported in the scientific literature on the topic that involve the cell wall, as a dynamic organelle, in cross-stress protection [51].…”

“…The important role played by the cell wall in yeast adaptation and tolerance to different stresses of biotechnological interest clearly emerges from a recent review article [51]. Results here described are in line with those reported in the scientific literature on the topic that involve the cell wall, as a dynamic organelle, in cross-stress protection [51]. However, most of the results on the topic reported so far have been dedicated to the model yeast and cell factory Saccharomyces cerevisiae being scarce and the available information concerning non-conventional yeast species of biotechnological relevance.…”

Sugar Utilization-Associated Food-Grade Selection Markers in Lactic Acid Bacteria and Yeast

“…As edible microorganisms, lactic acid bacteria and yeast have bright application prospects. Presently, many food-grade, genetically engineered microorganisms have been successfully constructed, and they have been applied in health care, fermentation, food processing, and other fields ( Abrha et al 2023 ; González-Orozco et al 2023 ; Van de Voorde et al 2023 ). The selection markers on vectors are an important part of the selection system for recombinant microorganisms.…”