Background: Xylose transport is one of the bottlenecks in the conversion of lignocellulosic biomass to ethanol. Xylose consumption by the wild-type strains of xylose-utilizing yeasts occurs once glucose is depleted resulting in a long fermentation process and overall slow and incomplete conversion of sugars liberated from lignocellulosic hydrolysates. Therefore, the engineering of endogenous transporters for the facilitation of glucose-xylose co-consumption is an important prerequisite for efficient ethanol production from lignocellulosic hydrolysates. Results: In this study, several engineering approaches formerly used for the low-affinity glucose transporters in Saccharomyces cerevisiae, were successfully applied for earlier identified transporter Hxt1 in Ogataea polymorpha to improve xylose consumption (engineering involved asparagine substitution to alanine at position 358 and replacement of N-terminal lysine residues predicted to be the target of ubiquitination for arginine residues). Moreover, the modified versions of S. cerevisiae Hxt7 and Gal2 transporters also led to improved xylose fermentation when expressed in O. polymorpha. Conclusions: The O. polymorpha strains with modified Hxt1 were characterized by simultaneous utilization of both glucose and xylose, in contrast to the wild-type and parental strain with elevated ethanol production from xylose. When the engineered Hxt1 transporter was introduced into constructed earlier advanced ethanol producer form xylose, the resulting strain showed further increase in ethanol accumulation during xylose fermentation. The overexpression of heterologous S. cerevisiae Gal2 had a less profound positive effects on sugars uptake rate, while overexpression of Hxt7 revealed the least impact on sugars consumption.
We synthesized a set of 13 new and earlier described styrylpyridinium compounds (N‐alkyl styrylpyridinium salts with bromide or tosylate anions) in order to evaluate antifungal activity against C. albicans cells, to assay the possible synergism with fluconazole, and to estimate cytotoxicity to mammalian cells. All compounds were synthesized according to a well‐known two‐step procedure involving alkylation of γ‐picoline with appropriate alkyl bromide and further condensation with substituted benzaldehyde. Compounds with long N‐alkyl chains (C18H37–C20H41) had no antifungal activity against the cells of all tested C. albicans strains. Other styrylpyridinium compounds were able to inhibit yeast growth at the concentrations of 0.06–16 μg/ml. At fungicidal concentrations, the compound with the CN‐ group was least toxic to mammalian cells, showed the most effective synergism with fluconazole, and only slightly inhibited the respiration of C. albicans. The compound with the 4′‐diethylamino group exhibited the strongest fungicidal properties and effectively blocked the respiration of C. albicans cells. However, toxicity to mammalian cells was also high. Summarizing, the results of our study indicate that styrylpyridinium compounds are promising candidates in the development of new antifungal drugs.
Water contamination by various bacteria, viruses and other pathogens is a great threat to human health. Amongst other Advanced Oxidation Processes TiO2 photocatalysis is considered as one of the most efficient treatment for the polluted wastewater disinfection. Usually, the wastewater produced by higher risk objects, such as hospitals, implicates diverse contaminants, but efficiency of most of the Advanced Oxidation Processes is tested by using only single pathogens and information on inactivation of bacteria mixtures is still limited. In this study, photocatalytical inactivation of three commonly found bacterial pathogens (gram-positive (Micrococcus luteus) and gram-negative (Salmonella enterica, Escherichia coli)) was investigated. Efficiency of traditional photocatalytic disinfection process using single bacterial pathogens was compared to the one observed for their mixtures. The impact of photocatalytical process parameters and treatment time on bacteria disinfection efficiency was studied. Photocatalytic disinfection efficiency testing with bacteria mixtures revealed, that in the presence of TiO2 photocatalyst and UV irradiation tested gram-positive cells were inactivated slower than gram-negative cells. Another important finding was that an overall photocatalytic disinfection efficiency of bacteria mixtures is not a straight forward sum of inactivation rates of individually tested pathogens but has a strong relationship to the properties of their competitive growth.
A lot of human activities have negative impact on water quality and sometimes result in the biological water contamination. Currently used chemical (chlorine, ozone, and etc.) and physical (UV) water disinfection methods have strong environmental disadvantages or suffers from limited efficiency. To overcome these problems, scientists suggest to use photocatalyst activated advanced oxidation processes. One of the most studied photocatalysts which attracts a lot of research interest is titanium dioxide. TiO2 application for the disinfection of water, air or surfaces is increasingly encouraged by researchers. However, to unlock its full potential it is highly desirable to make it suitable for the visible light activation. In the current study the effect of visible light assisted photocatalytic treatment to the outer membrane permeability of Salmonella enterica bacteria and how it changes under different titanium dioxide concentrations was analysed. The results from the treatment of relatively complex Salmonella enterica bacteria organism were compared to the visible light activated TiO2 ability to oxidise considerably simpler objects like methylene blue molecules. The efficiency of TiO2 photocatalytic disinfection process was evaluated using spread plate technique. Membrane permeability of the treated Salmonella enterica bacteria was determined by NPN uptake factor assay. Generation of intracellular reactive oxygen species was evaluated by Dichlorodihydrofluorescein diacetate fluorescence measurements. The key finding of this study was that intense wide spectrum visible light irradiation and TiO2 powder synergistically inactivate S. enterica bacteria and halt its potential to form colonies. High amounts of intracellular reactive oxygen species could be seen as the main suspects for the observed inactivation of S. enterica.
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