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1.
Widespread use of unconventional targeting signals in mitochondrial ribosome proteins.
Bykov, YS, Flohr, T, Boos, F, Zung, N, Herrmann, JM, Schuldiner, M
The EMBO journal. 2022;(1):e109519
Abstract
Mitochondrial ribosomes are complex molecular machines indispensable for respiration. Their assembly involves the import of several dozens of mitochondrial ribosomal proteins (MRPs), encoded in the nuclear genome, into the mitochondrial matrix. Proteomic and structural data as well as computational predictions indicate that up to 25% of yeast MRPs do not have a conventional N-terminal mitochondrial targeting signal (MTS). We experimentally characterized a set of 15 yeast MRPs in vivo and found that five use internal MTSs. Further analysis of a conserved model MRP, Mrp17/bS6m, revealed the identity of the internal targeting signal. Similar to conventional MTS-containing proteins, the internal sequence mediates binding to TOM complexes. The entire sequence of Mrp17 contains positive charges mediating translocation. The fact that these sequence properties could not be reliably predicted by standard methods shows that mitochondrial protein targeting is more versatile than expected. We hypothesize that structural constraints imposed by ribosome assembly interfaces may have disfavored N-terminal presequences and driven the evolution of internal targeting signals in MRPs.
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2.
Spent Brewer's Yeast as a Source of Insoluble β-Glucans.
Avramia, I, Amariei, S
International journal of molecular sciences. 2021;(2)
Abstract
In the brewing process, the consumption of resources and the amount of waste generated are high and due to a lot of organic compounds in waste-water, the capacity of natural regeneration of the environment is exceeded. Residual yeast, the second by-product of brewing is considered to have an important chemical composition. An approach with nutritional potential refers to the extraction of bioactive compounds from the yeast cell wall, such as β-glucans. Concerning the potential food applications with better textural characteristics, spent brewer's yeast glucan has high emulsion stability and water-holding capacity fitting best as a fat replacer in different food matrices. Few studies demonstrate the importance and nutritional role of β-glucans from brewer's yeast, and even less for spent brewer's yeast, due to additional steps in the extraction process. This review focuses on describing the process of obtaining insoluble β-glucans (particulate) from spent brewer's yeast and provides an insight into how a by-product from brewing can be converted to potential food applications.
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3.
Improvement of ethanol production in fed-batch fermentation using a mixture of sugarcane juice and molasse under very high-gravity conditions.
Cruz, ML, de Resende, MM, Ribeiro, EJ
Bioprocess and biosystems engineering. 2021;(3):617-625
Abstract
Ethanol fermentation in very high gravity (VHG) saves energy consumption for ethanol distillation. As the technology offers high ethanol yield and low waste generation and it can be operated at low cost, it could be more efficient at an industrial scale than other ethanol production methods. This work studied ethanol production using a fed-batch bioreactor with a working volume of 1.5 L. The main objective of this research was evaluate the effects of temperature, sugar concentration, and cellular concentration using a Central Composite Design (CCD). Experimental conditions were selected using the surface response technique obtained from the CCD, and the results were validated to test the reproducibility. The following operating conditions were selected: temperature of 27.0 °C, sugar concentration 300.0 g/L, and cell concentration 15.0% (v/v). Under these conditions, after 30 h of fermentation the ethanol concentration, productivity and yield were 135.0 g/L, 4.42 g/(L·h) and 90.0%, respectively. All sugar was completely consumed.
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4.
Hybrid neural network modeling and particle swarm optimization for improved ethanol production from cashew apple juice.
da Silva Pereira, A, Pinheiro, ÁDT, Rocha, MVP, Gonçalves, LRB, Cartaxo, SJM
Bioprocess and biosystems engineering. 2021;(2):329-342
Abstract
A hybrid neural model (HNM) and particle swarm optimization (PSO) was used to optimize ethanol production by a flocculating yeast, grown on cashew apple juice. HNM was obtained by combining artificial neural network (ANN), which predicted reaction specific rates, to mass balance equations for substrate (S), product and biomass (X) concentration, being an alternative method for predicting the behavior of complex systems. ANNs training was conducted using an experimental set of data of X and S, temperature and stirring speed. The HNM was statistically validated against a new dataset, being capable of representing the system behavior. The model was optimized based on a multiobjective function relating efficiency and productivity by applying the PSO. Optimal estimated conditions were: S0 = 127 g L-1, X0 = 5.8 g L-1, 35 °C and 111 rpm. In this condition, an efficiency of 91.5% with a productivity of 8.0 g L-1 h-1 was obtained at approximately 7 h of fermentation.
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5.
Modeling the Ethanol Tolerance of the Probiotic Yeast Saccharomyces cerevisiae var. boulardii CNCM I-745 for its Possible Use in a Functional Beer.
Ramírez-Cota, GY, López-Villegas, EO, Jiménez-Aparicio, AR, Hernández-Sánchez, H
Probiotics and antimicrobial proteins. 2021;(1):187-194
Abstract
Saccharomyces yeasts are able to ferment simple sugars to generate levels of ethanol that are toxic to other yeasts and bacteria. The tolerance to ethanol of different yeasts depends also on the incubation temperature. In this study, the ethanol stress responses of S. cerevisiae and the probiotic yeast S. boulardii CNCM I-745 were evaluated at two temperatures. The growth kinetics parameters were obtained by fitting the Baranyi and Roberts model to the experimental data. The four-parameter logistic Hill equation was used to describe the ethanol tolerance of the yeasts at the temperatures of 28 and 37 °C. Adequate determination coefficients were obtained (R2 > 0.91) in all cases. S. boulardii grown at 28 °C was selected as the yeast with the best ethanol tolerance (6-8%) for use in the elaboration of functional craft beers.
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6.
Copper metabolism in Saccharomyces cerevisiae: an update.
Shi, H, Jiang, Y, Yang, Y, Peng, Y, Li, C
Biometals : an international journal on the role of metal ions in biology, biochemistry, and medicine. 2021;(1):3-14
Abstract
Copper is an essential element in all forms of life. It acts as a cofactor of some enzymes and is involved in forming proper protein conformations. However, excess copper ions in cells are detrimental as they can generate free radicals or disrupt protein structures. Therefore, all life forms have evolved conserved and exquisite copper metabolic systems to maintain copper homeostasis. The yeast Saccharomyces cerevisiae has been widely used to investigate copper metabolism as it is convenient for this purpose. In this review, we summarize the mechanism of copper metabolism in Saccharomyces cerevisiae according to the latest literature. In brief, bioavailable copper ions are incorporated into yeast cells mainly via the high-affinity transporters Ctr1 and Ctr3. Then, intracellular Cu+ ions are delivered to different organelles or cuproproteins by different chaperones, including Ccs1, Atx1, and Cox17. Excess copper ions bind to glutathione (GSH), metallothioneins, and copper complexes are sequestered into vacuoles to avoid toxicity. Copper-sensing transcription factors Ace1 and Mac1 regulate the expression of genes involved in copper detoxification and uptake/mobilization in response to changes in intracellular copper levels. Though numerous recent breakthroughs in understanding yeast's copper metabolism have been achieved, some issues remain unresolved. Completely elucidating the mechanism of copper metabolism in yeast helps decode the corresponding system in humans and understand how copper-related diseases develop.
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7.
Pib2 as an Emerging Master Regulator of Yeast TORC1.
Hatakeyama, R
Biomolecules. 2021;(10)
Abstract
Cell growth is dynamically regulated in response to external cues such as nutrient availability, growth factor signals, and stresses. Central to this adaptation process is the Target of Rapamycin Complex 1 (TORC1), an evolutionarily conserved kinase complex that fine-tunes an enormous number of cellular events. How upstream signals are sensed and transmitted to TORC1 has been intensively studied in major model organisms including the budding yeast Saccharomyces cerevisiae. This field recently saw a breakthrough: the identification of yeast phosphatidylInositol(3)-phosphate binding protein 2 (Pib2) protein as a critical regulator of TORC1. Although the study of Pib2 is still in its early days, multiple groups have provided important mechanistic insights on how Pib2 relays nutrient signals to TORC1. There remain, on the other hand, significant gaps in our knowledge and mysteries that warrant further investigations. This is the first dedicated review on Pib2 that summarizes major findings and outstanding questions around this emerging key player in cell growth regulation.
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8.
Lipid Droplets and Their Autophagic Turnover via the Raft-Like Vacuolar Microdomains.
Rahman, MA, Kumar, R, Sanchez, E, Nazarko, TY
International journal of molecular sciences. 2021;(15)
Abstract
Although once perceived as inert structures that merely serve for lipid storage, lipid droplets (LDs) have proven to be the dynamic organelles that hold many cellular functions. The LDs' basic structure of a hydrophobic core consisting of neutral lipids and enclosed in a phospholipid monolayer allows for quick lipid accessibility for intracellular energy and membrane production. Whereas formed at the peripheral and perinuclear endoplasmic reticulum, LDs are degraded either in the cytosol by lipolysis or in the vacuoles/lysosomes by autophagy. Autophagy is a regulated breakdown of dysfunctional, damaged, or surplus cellular components. The selective autophagy of LDs is called lipophagy. Here, we review LDs and their degradation by lipophagy in yeast, which proceeds via the micrometer-scale raft-like lipid domains in the vacuolar membrane. These vacuolar microdomains form during nutrient deprivation and facilitate internalization of LDs via the vacuolar membrane invagination and scission. The resultant intra-vacuolar autophagic bodies with LDs inside are broken down by vacuolar lipases and proteases. This type of lipophagy is called microlipophagy as it resembles microautophagy, the type of autophagy when substrates are sequestered right at the surface of a lytic compartment. Yeast microlipophagy via the raft-like vacuolar microdomains is a great model system to study the role of lipid domains in microautophagic pathways.
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9.
Multi-omics based strategy for toxicity analysis of acrylamide in Saccharomyces cerevisiae model.
Lin, Z, Luo, P, Huang, D, Wu, Y, Li, F, Liu, H
Chemico-biological interactions. 2021;:109682
Abstract
Although the toxicity of acrylamide (ACR) has been extensively investigated in different experimental models, its perturbations to multiple nodes of the cellular signaling network have not been systematically associated. In this study, changes at different omics layers in ACR exposed Saccharomyces cerevisiae cells were monitored using a multi-omics strategy. The results of the analysis highlighted the impairment of oxidative-reductive balance, energy metabolism, lipid metabolism, nucleotide metabolism, and ribosome function in yeast cells. Response to acute ACR damage, glutathione synthesis was upregulated, the process of protein degradation was accelerated, and the autophagy flux was initiated. Meanwhile, yeast upregulates gene expression levels of enzymes in carbohydrate metabolism and speeds up the oxidation process of fatty acids to compensate for energy depletion. Importantly, the multi-omics strategy captures features that have rarely been addressed in previous studies on the toxicology of ACR, including blocked de novo nucleotide synthesis, decreased levels of metabolic enzyme cofactors thiamine and D-biotin, increased intracellular concentrations of neurotoxic N-methyl d-aspartic acid and l-glutamic acid, and release of death mediators ceramide. The ACR perturbation network constructed in this work and the discovery of new damage features provide a theoretical basis for subsequent point-to-point toxicological studies.
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10.
Modeling and analysis of the macronutrient signaling network in budding yeast.
Jalihal, AP, Kraikivski, P, Murali, TM, Tyson, JJ
Molecular biology of the cell. 2021;(21):ar20
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Abstract
Adaptive modulation of the global cellular growth state of unicellular organisms is crucial for their survival in fluctuating nutrient environments. Because these organisms must be able to respond reliably to ever varying and unpredictable nutritional conditions, their nutrient signaling networks must have a certain inbuilt robustness. In eukaryotes, such as the budding yeast Saccharomyces cerevisiae, distinct nutrient signals are relayed by specific plasma membrane receptors to signal transduction pathways that are interconnected in complex information-processing networks, which have been well characterized. However, the complexity of the signaling network confounds the interpretation of the overall regulatory "logic" of the control system. Here, we propose a literature-curated molecular mechanism of the integrated nutrient signaling network in budding yeast, focusing on early temporal responses to carbon and nitrogen signaling. We build a computational model of this network to reconcile literature-curated quantitative experimental data with our proposed molecular mechanism. We evaluate the robustness of our estimates of the model's kinetic parameter values. We test the model by comparing predictions made in mutant strains with qualitative experimental observations made in the same strains. Finally, we use the model to predict nutrient-responsive transcription factor activities in a number of mutant strains undergoing complex nutrient shifts.