Metallothioneins, Saccharomyces cerevisiae, and Heavy Metals: A Biotechnology Triad?

Metal ions are the least sophisticated chemical species that interact or bind to biomolecules. The yeast Saccharomyces cerevisiae represents a versatile model organisms used in both basic and applicative research, and one of the main contributors to the understanding of the molecular mechanisms involved in the transport, accumulation, and homeostasis of heavy metals. With a negatively charged wall, the yeast cells are very good biosorbents for heavy metals. In addition to biosorption, the metabolically active cells take up heavy metals via the normal membrane transport systems. Once in the cell, the toxicity of the heavy metals is controlled by various mechanisms, including sequestration by metal-binding proteins, such as the metallothioneins. Metallothioneins are cysteine-rich proteins involved in the buffering of excess heavy metals, both essential (Cu and Zn) and nonessential (Cd, Ag, and Hg). S. cerevisiae has two innate metallothioneins, Cup1 and Crs5, intensively investigated. Additionally, S. cerevisiae served as a host for the heterologous expression of a variety of metallothioneins from different species. This review focuses on the technological implications of expressing metallothioneins in yeast and on the possibility to use these transgenic cells in heavy metal-related biotechnologies: bioremediation, recovery of rare metals, or obtaining clonable tags for protein imaging

Calcium and Cell Response to Heavy Metals: Can Yeast Provide an Answer?

Despite constant efforts to maintain a clean environment, heavy metal pollution continues to raise challenges to the industrialized world. Exposure to heavy metals is detrimental to living organisms, and it is of utmost importance that cells find rapid and efficient ways to respond to and eventually adapt to surplus metals for survival under severe stress. This chapter focuses on the attempts done so far to elucidate the calcium-mediated response to heavy metal stress using the model organism Saccharomyces cerevisiae. The possibilities to record the transient elevations of calcium within yeast cells concomitantly with the heavy metal exposure are presented, and the limitations imposed by interference between calcium and heavy metals are discussed

Optical manipulation of Saccharomyces cerevisiae cells reveals that green light protection against UV irradiation is favored by low Ca2+ and requires intact UPR pathway

AbstractOptical manipulation of Saccharomyces cerevisiae cells with high density green photons conferred protection against the deleterious effects of UV radiation. Combining chemical screening with UV irradiation of yeast cells, it was noted that the high density green photons relied on the presence of intact unfolded protein response (UPR) pathway to exert their protective effect and that the low Ca2+ conditions boosted the effect. UPR chemical inducers tunicamycin, dithiotreitol and calcium chelators augmented the green light effect in a synergic action against UV-induced damage. Photo-manipulation of cells was a critical factor since the maximum protection was achieved only when cells were pre-exposed to green light

Saccharomyces cerevisiae and Caffeine Implications on the Eukaryotic Cell

Caffeine–a methylxanthine analogue of the purine bases adenine and guanine–is by far the most consumed neuro-stimulant, being the active principle of widely consumed beverages such as coffee, tea, hot chocolate, and cola. While the best-known action of caffeine is to prevent sleepiness by blocking the adenosine receptors, caffeine exerts a pleiotropic effect on cells, which lead to the activation or inhibition of various cell integrity pathways. The aim of this review is to present the main studies set to investigate the effects of caffeine on cells using the model eukaryotic microorganism Saccharomyces cerevisiae, highlighting the caffeine synergy with external cell stressors, such as irradiation or exposure to various chemical hazards, including cigarette smoke or chemical carcinogens. The review also focuses on the importance of caffeine-related yeast phenotypes used to resolve molecular mechanisms involved in cell signaling through conserved pathways, such as target of rapamycin (TOR) signaling, Pkc1-Mpk1 mitogen activated protein kinase (MAPK) cascade, or Ras/cAMP protein kinase A (PKA) pathway

Coffee and Yeasts: From Flavor to Biotechnology

Coffee is one of the most consumed beverages in the world, and its popularity has prompted the necessity to constantly increase the variety and improve the characteristics of coffee as a general commodity. The popularity of coffee as a staple drink has also brought undesired side effects, since coffee production, processing and consumption are all accompanied by impressive quantities of coffee-related wastes which can be a threat to the environment. In this review, we integrated the main studies on fermentative yeasts used in coffee-related industries with emphasis on two different directions: (1) the role of yeast strains in the postharvest processing of coffee, the possibilities to use them as starting cultures for controlled fermentation and their impact on the sensorial quality of processed coffee, and (2) the potential to use yeasts to capitalize on coffee wastes—especially spent coffee grounds—in the form of eco-friendly biomass, biofuel or fine chemical production

Anthocyanins and Anthocyanin-Derived Products in Yeast-Fermented Beverages

The beverages obtained by yeast fermentation from anthocyanin-rich natural sources (grapes, berries, brown rice, etc.) retain part of the initial pigments in the maturated drink. During the fermentation and aging processes anthocyanins undergo various chemical transformations, which include reactions with glycolytic products (especially pyruvate and acetaldehyde) or with other compounds present in the complex fermentation milieu (such as vinylphenols obtained from cinnamic acids by means of a yeast decarboxylase) yielding pigments which can be more stable than the initial anthocyanins. Overall, these compounds contribute to the organoleptic traits of the mature product, but also to the overall chemical composition which make the yeast fermented beverages important sources of dietary antioxidants. In this review, we focused on the studies regarding the changes underwent by anthocyanins during yeast-mediated fermentation, on the approaches taken to enrich the fermented beverages in anthocyanins and their derived products, and on the interrelations between yeast and anthocyanin which were of relevance for obtaining a high-quality product containing optimum amounts of anthocyanin and anthocyanin-derived products

Kcs1 and Vip1: The Key Enzymes behind Inositol Pyrophosphate Signaling in <i>Saccharomyces cerevisiae</i>

The inositol pyrophosphate pathway, a complex cell signaling network, plays a pivotal role in orchestrating vital cellular processes in the budding yeast, where it regulates cell cycle progression, growth, endocytosis, exocytosis, apoptosis, telomere elongation, ribosome biogenesis, and stress responses. This pathway has gained significant attention in pharmacology and medicine due to its role in generating inositol pyrophosphates, which serve as crucial signaling molecules not only in yeast, but also in higher eukaryotes. As targets for therapeutic development, genetic modifications within this pathway hold promise for disease treatment strategies, offering practical applications in biotechnology. The model organism Saccharomyces cerevisiae, renowned for its genetic tractability, has been instrumental in various studies related to the inositol pyrophosphate pathway. This review is focused on the Kcs1 and Vip1, the two enzymes involved in the biosynthesis of inositol pyrophosphate in S. cerevisiae, highlighting their roles in various cell processes, and providing an up-to-date overview of their relationship with phosphate homeostasis. Moreover, the review underscores the potential applications of these findings in the realms of medicine and biotechnology, highlighting the profound implications of comprehending this intricate signaling network

Nanobodies as Diagnostic and Therapeutic Tools for Cardiovascular Diseases (CVDs)

The aim of this review is to summarize some of the most recent work in the field of cardiovascular disease (CVD) diagnosis and therapy, focusing mainly on the role of nanobodies in the development of non-invasive imaging methods, diagnostic devices, and advanced biotechnological therapy tools. In the context of the increased number of people suffering from CVDs due to a variety of factors such as sedentariness, poor nutrition, stress, and smoking, there is an urgent need for new and improved diagnostic and therapeutic methods. Nanobodies can be easily produced in prokaryotes, lower eukaryotes, and plant and mammalian cells, and offer great advantages. In the diagnosis domain, they are mainly used as labeled probes that bind to certain surface receptors or other target molecules and give important information on the severity and extent of atherosclerotic lesions, using imaging methods such as contrast-enhanced ultrasound molecular imaging (CEUMI), positron emission tomography (PET), single-photon emission computed tomography coupled with computed tomography (SPECT/CT), and PET/CT. As therapy tools, nanobodies have been used either for transporting drug-loaded vesicles to specific targets or as inhibitors for certain enzymes and receptors, demonstrated to be involved in various CVDs

Anchoring plant metallothioneins to the inner face of the plasma membrane of Saccharomyces cerevisiae cells leads to heavy metal accumulation.

In this study we engineered yeast cells armed for heavy metal accumulation by targeting plant metallothioneins to the inner face of the yeast plasma membrane. Metallothioneins (MTs) are cysteine-rich proteins involved in the buffering of excess metal ions, especially Cu(I), Zn(II) or Cd(II). The cDNAs of seven Arabidopsis thaliana MTs (AtMT1a, AtMT1c, AtMT2a, AtMT2b, AtMT3, AtMT4a and AtMT4b) and four Noccaea caerulescens MTs (NcMT1, NcMT2a, NcMT2b and NcMT3) were each translationally fused to the C-terminus of a myristoylation green fluorescent protein variant (myrGFP) and expressed in Saccharomyces cerevisiae cells. The myrGFP cassette introduced a yeast myristoylation sequence which allowed directional targeting to the cytosolic face of the plasma membrane along with direct monitoring of the intracellular localization of the recombinant protein by fluorescence microscopy. The yeast strains expressing plant MTs were investigated against an array of heavy metals in order to identify strains which exhibit the (hyper)accumulation phenotype without developing toxicity symptoms. Among the transgenic strains which could accumulate Cu(II), Zn(II) or Cd(II), but also non-canonical metal ions, such as Co(II), Mn(II) or Ni(II), myrGFP-NcMT3 qualified as the best candidate for bioremediation applications, thanks to the robust growth accompanied by significant accumulative capacity