Biosensors are low‐cost and low‐maintenance alternatives to conventional analytical techniques for biomedical, industrial and environmental applications. Biosensors based on whole microorganisms can be genetically engineered to attain high sensitivity and specificity for the detection of selected analytes. While bacteria‐based biosensors have been extensively reported, there is a recent interest in yeast‐based biosensors, combining the microbial with the eukaryotic advantages, including possession of specific receptors, stability and high robustness. Here, we describe recently reported yeast‐based biosensors highlighting their biological and technical features together with their status of development, that is, laboratory or prototype. Notably, most yeast‐based biosensors are still in the early developmental stage, with only a few prototypes tested for real applications. Open challenges, including systematic use of advanced molecular and biotechnological tools, bioprospecting, and implementation of yeast‐based biosensors in electrochemical setup, are discussed to find possible solutions for overcoming bottlenecks and promote real‐world application of yeast‐based biosensors.

Mitochondrial RTG (an acronym for ReTroGrade) signaling plays a cytoprotective role under various intracellular or environmental stresses. We have previously shown its contribution to osmoadaptation and capacity to sustain mitochondrial respiration in yeast. Here, we studied the interplay between RTG2, the main positive regulator of the RTG pathway, and HAP4, encoding the catalytic subunit of the Hap2-5 complex required for the expression of many mitochondrial proteins that function in the tricarboxylic acid (TCA) cycle and electron transport, upon osmotic stress. Cell growth features, mitochondrial respiratory competence, retrograde signaling activation, and TCA cycle gene expression were comparatively evaluated in wild type and mutant cells in the presence and in the absence of salt stress. We showed that the inactivation of HAP4 improved the kinetics of osmoadaptation by eliciting both the activation of retrograde signaling and the upregulation of three TCA cycle genes: citrate synthase 1 (CIT1), aconitase 1 (ACO1), and isocitrate dehydrogenase 1 (IDH1). Interestingly, their increased expression was mostly dependent on RTG2. Impaired respiratory competence in the HAP4 mutant does not affect its faster adaptive response to stress. These findings indicate that the involvement of the RTG pathway in osmostress is fostered in a cellular context of constitutively reduced respiratory capacity. Moreover, it is evident that the RTG pathway mediates peroxisomes–mitochondria communication by modulating the metabolic function of mitochondria in osmoadaptation.

Electrodes functionalised with weak electroactive microorganisms offer a viable alternative to conventional chemical sensors for detecting priority pollutants in bioremediation processes. Biofilm-based biosensors have been proposed for this purpose. However, biofilm formation and maturation require 24–48 h, and the microstructure and coverage of the electrode surface cannot be controlled, leading to poorly reproducible signal and sensitivity. Alternatively, semiconductive biocompatible coatings can be used for viable cell immobilization, achieving reproducible coverage and resulting in a stable biosensor response. In this work, we use a polydopamine (PDA)-based coating to immobilize Saccharomyces cerevisiae yeast viable cells on carbon screen printed electrodes (SPE) for Cu(II) detection, with potassium ferricyanide (K₃[Fe (CN)₆]) as a redox mediator. Under these conditions, the current output correlates with Cu (II) concentration, reaching a limit of detection of 2.2 µM, as calculated from the chronoamperometric response. The bioelectrochemical results are supported by standard viability assays, microscopy, and electrochemical impedance spectroscopy. The PDA coatings can be functionalised with different mutant strains, thus expanding the toolbox for biosensor design in bioremediation.

The coupling of biological organisms with electrodes enables the development of sustainable, low cost, and potentially self-sustained biosensors. A critical aspect is to obtain portable bioelectrodes where the biological material is immobilized on the electrode surface to be utilized on demand. Herein, we developed an approach for the rapid entrapment and immobilization of metabolically active yeast cells in a biocompatible polydopamine layer, which does not require a separate and time-consuming synthesis. The reported approach allows obtaining the "electrical wire" of intact and active yeast cells with resulting current generation from glucose oxidation. Additionally, the electrochemical performance of the biohybrid yeast-based system has been characterized in the presence of CuSO₄, a widely used pesticide, in the environmentally relevant concentration range of 20-100 μM. The system enabled the rapid preliminary monitoring of the contaminant based on variations in current generation, with a limit of detection of 12.5 μM CuSO₄. The present approach for the facile preparation of portable yeast-based electrochemical biosensors paves the way for the future development of sustainable systems for environmental monitoring.

Copper contamination in the environment poses significant risks to both soil and human health, making the need for reliable monitoring methods crucial. In this study, we report the use of the EmStat Pico module as potentiostat to develop a portable electrochemical biosensor for copper detection, utilizing yeast Saccharomyces cerevisiae cells immobilized on a polydopamine (PDA)-coated screen-printed electrode (SPE). By optimizing the sensor design with a horizontal assembly and the volume reduction in the electrolyte solution, we achieved a 10-fold increase in current density with higher range of copper concentrations (0–300 µM CuSO₄) compared to traditional (or previous) vertical dipping setups. Additionally, the use of genetically engineered copper-responsive yeast cells further improved sensor performance, with the recombinant strain showing a 1.7-fold increase in current density over the wild-type strain. The biosensor demonstrated excellent reproducibility (R² > 0.95) and linearity over a broad range of copper concentrations, making it suitable for precise quantitative analysis. To further enhance portability and usability, a Bluetooth-enabled electrochemical platform was integrated with a web application for real-time data analysis, enabling on-site monitoring and providing a reliable, cost-effective tool for copper detection in real world settings. This system offers a promising solution for addressing the growing need for efficient environmental monitoring, especially in agriculture.

Quantitative assessment of growth and survival is a suitable technique in studying biochemical, genetic and physiological processes in the cells. The budding yeast Saccharomyces cerevisiae is one of the most widely used eukaryotic model organisms for studying cellular mechanisms and processes in evolutionarily distant species, including humans. Yeast growth can be evaluated on both liquid and solid media by measuring cell suspension turbidity and colony forming units, respectively. Several software tools utilizing different parameters have been proposed to quantify yeast growth on solid media. Here, we developed a Matlab-based application which provides a rapid and robust quantitative yeast growth analysis from spot plating assay. Spot plating assay is a typical procedure to evaluate yeast growth in low-throughput laboratory settings, including growth on different nutrient sources or treatment with specific stressors. The app has a one-step installation process, a self-explanatory interface and shorter analysis steps compared with previous established methods, providing a useful tool for both expert and non-expert yeast researchers.