In recent years, many novel xylose-fermenting yeasts belonging to the new genus Spathaspora have been isolated from the gut of wood-feeding insects and/or wood-decaying substrates. We have cloned and expressed, in Saccharomyces cerevisiae, a Spathaspora arborariae xylose reductase gene (SaXYL1) that accepts both NADH and NADPH as co-substrates, as well as a Spathaspora passalidarum NADPH-dependent xylose reductase (SpXYL1.1 gene) and the SpXYL2.2 gene encoding for a NAD+-dependent xylitol dehydrogenase. These enzymes were co-expressed in a S. cerevisiae strain over-expressing the native XKS1 gene encoding xylulokinase, as well as being deleted in the alkaline phosphatase encoded by the PHO13 gene. The S. cerevisiae strains expressing the Spathaspora enzymes consumed xylose, and xylitol was the major fermentation product. Higher specific growth rates, xylose consumption and xylitol volumetric productivities were obtained by the co-expression of the SaXYL1 and SpXYL2.2 genes, when compared with the co-expression of the NADPH-dependent SpXYL1.1 xylose reductase. During glucose-xylose co-fermentation by the strain with co-expression of the SaXYL1 and SpXYL2.2 genes, both ethanol and xylitol were produced efficiently. Our results open up the possibility of using the advantageous Saccharomyces yeasts for xylitol production, a commodity with wide commercial applications in pharmaceuticals, nutraceuticals, food and beverage industries.
Yeasts are the most used microorganisms for biotechnological purposes. Although they have been mainly recognized for their application in the beverage and bioethanol industries, these microorganisms can be efficiently employed in pharmaceutical and food production companies. In these industrial sectors, yeasts are highly desirable for their capacity to produce bioactive compounds from simple substrates, including wastes. In this review, we present the state of the art of bioactive compound production in microbial cell factories and analyze the avenues to increase the productivity of these molecules, which benefit human and environmental health. The article addresses their vast biological activities, from preventing to treating human diseases and from pre to postharvest control on agroindustrial streams. Furthermore, different yeast species, genetically engineered or not, are herein presented not only as biofactories of the referred to compounds but also as their targets. This comprehensive analysis of the literature points out the significant roles of biodiversity, bioprospection, and genome editing tools on the microbial production of bioactive compounds and reveals the value of these approaches from the one health perspective.
The literature is full of studies reporting environmental and health issues related to using traditional pesticides in food production and storage. Fortunately, alternatives have arisen in the last few decades, showing that organic agriculture is possible and economically feasible. And in this scenario, fungi may be helpful. In the natural environment, when associated with plants, these microorganisms offer plant-growth-promoting molecules, facilitate plant nutrient uptake, and antagonize phytopathogens. It is true that fungi can also be phytopathogenic, but even they can benefit agriculture in some way—since pathogenicity is species-specific, these fungi are shown to be useful against weeds (as bioherbicides). Finally, plant-associated yeasts and molds are natural biofactories, and the metabolites they produce while dwelling in leaves, flowers, roots, or the rhizosphere have the potential to be employed in different industrial activities. By addressing all these subjects, this manuscript comprehensively reviews the biotechnological uses of plant-associated fungi and, in addition, aims to sensitize academics, researchers, and investors to new alternatives for healthier and more environmentally friendly production processes.
In previous work, we developed a Saccharomyces cerevisiae strain (DLG-K1) lacking the main monosaccharide transporters (hxt-null) and displaying high xylose reductase, xylitol dehydrogenase and xylulokinase activities. This strain proved to be a useful chassis strain to study new glucose/xylose transporters, as SsXUT1 from Scheffersomyces stipitis. Proteins with high amino acid sequence similarity (78–80%) to SsXUT1 were identified from Spathaspora passalidarum and Spathaspora arborariae genomes. The characterization of these putative transporter genes (SpXUT1 and SaXUT1, respectively) was performed in the same chassis strain. Surprisingly, the cloned genes could not restore the ability to grow in several monosaccharides tested (including glucose and xylose), but after being grown in maltose, the uptake of 14C-glucose and 14C-xylose was detected. While SsXUT1 lacks lysine residues with high ubiquitinylation potential in its N-terminal domain and displays only one in its C-terminal domain, both SpXUT1 and SaXUT1 transporters have several such residues in their C-terminal domains. A truncated version of SpXUT1 gene, deprived of the respective 3′-end, was cloned in DLG-K1 and allowed growth and fermentation in glucose or xylose. In another approach, two arrestins known to be involved in the ubiquitinylation and endocytosis of sugar transporters (ROD1 and ROG3) were knocked out, but only the rog3 mutant allowed a significant improvement of growth and fermentation in glucose when either of the XUT permeases were expressed. Therefore, for the efficient heterologous expression of monosaccharide (e.g., glucose/xylose) transporters in S. cerevisiae, we propose either the removal of lysines involved in ubiquitinylation and endocytosis or the use of chassis strains hampered in the specific mechanism of membrane protein turnover.
Synthetic herbicides such as glyphosate and 2,4-D are widely used in agriculture but can negatively impact non-target organisms, including microorganisms essential for ecological balance. Yeasts associated with pollinating insects play crucial roles in plant–insect interactions, yet their responses to herbicides remain understudied. This study aimed to evaluate the capacity of yeasts isolated from bees and beetles to produce indole-3-acetic acid (IAA), a plant-growth-promoting hormone, as well as their ability to tolerate or degrade glyphosate (in the commercial herbicide Zapp QI 620®) and 2,4-D (in the commercial Aminol 806®). Seven yeast strains were isolated from insects, identified via ITS sequencing, and assessed for IAA production in YPD medium. Growth assays were conducted under varying herbicide concentrations, and 2,4-D degradation was analyzed using high-performance liquid chromatography. All strains produced IAA, with Papiliotrema siamensis CHAP-239 exhibiting the highest yield (4.17 mg/L). Glyphosate completely inhibited growth in all strains, while 2,4-D showed dose-dependent effects, with four strains tolerating lower concentrations. Notably, Meyerozyma caribbica CHAP-248 degraded up to 46% of 2,4-D at 6.045 g/L. These findings highlight the ecological risks herbicides pose to beneficial yeasts and suggest the potential of certain strains for bioremediation in herbicide-contaminated environments. Overall, the study underscores the importance of preserving microbial biodiversity in the context of sustainable agriculture.
Plant-associated yeasts play significant ecological roles within the microbiomes of soils and pollinating insects. In previous studies, we have shown that yeasts can assist pollinators in locating nectar, which is crucial for their nutrition and the reproduction of many angiosperms. Additionally, in soil, yeasts can also act as plant growth promoters. Given the importance of yeasts for plant development, this review first explores the biochemical processes underlying the ecological role of these microorganisms in soil, insects, and in direct association with plants. Based on this premise, we discuss the influence of these relationships on agricultural production, the biological mechanisms through which pesticides negatively affect yeast cells, and how these microorganisms can tolerate widely used agrochemicals. Finally, we address key studies in the literature that support the potential of these microorganisms as bioremediation agents. In this context, we emphasize different experiences with both indigenous and genetically engineered yeasts, which may display enzymes in their surfaces that convert pesticides into less harmful or nontoxic molecules. Our review indicates that yeasts can be effectively harnessed in organic agriculture to promote plant growth and bioremediate contaminated soil or food.