Lignocellulosic biomass is a promising source of renewable biofuels. However, pretreatment of lignocellulosic biomass generates fermentation inhibitors that adversely affect the growth of industrial microorganisms such as Saccharomyces cerevisiae and prevent economic production of lignocellulosic biofuels. A critical challenge on developing S. cerevisiae with improved inhibitor resistance lies in incomplete understanding of molecular basis for inhibitor stress response and limited information on effective genetic targets for increasing yeast resistance to mixed fermentation inhibitors. In this study, we applied comparative transcriptomic analysis to determine the molecular basis for acetic acid and/or furfural resistance in S. cerevisiae. We recently developed a yeast strain YC1 with superior resistance to acetic acid, furfural, and their mixture through inverse metabolic engineering. In this study, we first determined transcriptional changes through RNA sequencing in YC1 versus the wild-type strain S-C1 under three different inhibitor conditions, including acetic acid alone, furfural alone, and mixture of acetic acid and furfural. The genes associated with stress responses of S. cerevisiae to single and mixed inhibitors were revealed. Specifically, we identified 184 consensus genes that were differentially regulated in response to the distinct inhibitor resistance between YC1 and S-C1. Bioinformatic analysis next revealed key transcription factors (TFs) that regulate these consensus genes. The top TFs identified, Sfp1p and Ace2p, were experimentally tested as overexpression targets for strain optimization. Overexpression of the SFP1 gene improved specific ethanol productivity by nearly four times, while overexpression of the ACE2 gene enhanced the rate by three times in the presence of acetic acid and furfural. Overexpression of SFP1 gene in the resistant strain YC1 further resulted in 42 % increase in ethanol productivity in the presence of acetic acid and furfural, suggesting the effect of Sfp1p in optimizing the yeast strain for improved tolerance to mixed fermentation inhibitor. Transcriptional regulation underlying yeast resistance to acetic acid and furfural was determined. Two transcription factors, Sfp1p and Ace2p, were uncovered for the first time for their functions in improving yeast resistance to mixed fermentation inhibitors. The study demonstrated an omics-guided metabolic engineering framework, which could be developed as a promising strategy to improve complex microbial phenotypes.

Per- and polyfluoroalkyl substances (PFAS) make up a diverse group of industrially derived organic chemicals that are of significant concern due to their detrimental effects on human health and ecosystems. Although other technologies are available for removing PFAS, adsorption remains a viable and effective method. Accordingly, the current study reported a novel type of graphene oxide (GO)-based adsorbent and tested their removal performance toward removing PFAS from water. Among the eight adsorbents tested, GO modified by a cationic surfactant, cetyltrimethylammonium chloride (CTAC), GO-CTAC was found to be the best, showing an almost 100% removal for all 11 PFAS tested. The adsorption kinetics were best described by the pseudo-second-order model, indicating rapid adsorption. The isotherm data were well supported by the Toth model, suggesting that PFAS adsorption onto GO-CTAC involved complex interactions. Detailed characterization using scanning electron microscopy-energy dispersive X-ray spectroscopy, Fourier transform infrared, thermogravimetric analysis, X-ray diffraction, and X-ray photoelectron spectroscopy confirmed the proposed adsorption mechanisms, including electrostatic and hydrophobic interactions. Interestingly, the performance of GO-CTAC was not influenced by the solution pH, ionic strength, or natural organic matter. Furthermore, the removal efficiency of PFAS at almost 100% in river water demonstrated that GO-CTAC could be a suitable adsorbent for capturing PFAS in real surface water.

Single-walled carbon nanotubes (SWCNTs) have caused increasing public concerns associated with their potential toxicological effects. This study demonstrated that the physicochemical properties of SWCNTs have impacts on their toxicological effects.