scholarly journals Engineering energetically efficient transport of dicarboxylic acids in yeast Saccharomyces cerevisiae

2019 ◽  
Vol 116 (39) ◽  
pp. 19415-19420 ◽  
Author(s):  
Behrooz Darbani ◽  
Vratislav Stovicek ◽  
Steven Axel van der Hoek ◽  
Irina Borodina
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dicarboxylic Acid ◽  
Acid Production ◽  
Dicarboxylic Acids ◽  
Scale Production ◽  
Protein Motif ◽  
Yeast Saccharomyces Cerevisiae ◽  
Organic Acid Production ◽  
Cell Factories ◽  
Voltage Dependent

Biobased C4-dicarboxylic acids are attractive sustainable precursors for polymers and other materials. Commercial scale production of these acids at high titers requires efficient secretion by cell factories. In this study, we characterized 7 dicarboxylic acid transporters in Xenopus oocytes and in Saccharomyces cerevisiae engineered for dicarboxylic acid production. Among the tested transporters, the Mae1(p) from Schizosaccharomyces pombe had the highest activity toward succinic, malic, and fumaric acids and resulted in 3-, 8-, and 5-fold titer increases, respectively, in S. cerevisiae, while not affecting growth, which was in contrast to the tested transporters from the tellurite-resistance/dicarboxylate transporter (TDT) family or the Na+ coupled divalent anion–sodium symporter family. Similar to SpMae1(p), its homolog in Aspergillus carbonarius, AcDct(p), increased the malate titer 12-fold without affecting the growth. Phylogenetic and protein motif analyses mapped SpMae1(p) and AcDct(p) into the voltage-dependent slow-anion channel transporter (SLAC1) clade of transporters, which also include plant Slac1(p) transporters involved in stomata closure. The conserved phenylalanine residue F329 closing the transport pore of SpMae1(p) is essential for the transporter activity. The voltage-dependent SLAC1 transporters do not use proton or Na+ motive force and are, thus, less energetically expensive than the majority of other dicarboxylic acid transporters. Such transporters present a tremendous advantage for organic acid production via fermentation allowing a higher overall product yield.

scholarly journals Unique genetic basis of the distinct antibiotic potency of high acetic acid production in the probiotic yeast Saccharomyces cerevisiae var. boulardii

2019 ◽  
Vol 29 (9) ◽  
pp. 1478-1494 ◽  
Author(s):  
Benjamin Offei ◽  
Paul Vandecruys ◽  
Stijn De Graeve ◽  
María R. Foulquié-Moreno ◽  
Johan M. Thevelein
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Acetic Acid ◽  
Acid Production ◽  
Genetic Basis ◽  
Yeast Saccharomyces Cerevisiae ◽  
Acetic Acid Production ◽  
Probiotic Yeast

scholarly journals Analysis of lipid composition reveals mechanisms of ethanol tolerance in the model yeast Saccharomyces cerevisiae

2021 ◽  
Author(s):  
M Lairón-Peris ◽  
S. J. Routledge ◽  
J. A. Linney ◽  
J Alonso-del-Real ◽  
C.M. Spickett ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Stress Responses ◽  
Ethanol Tolerance ◽  
Yeast Species ◽  
Culture Media ◽  
Sugar Content ◽  
Yeast Cells ◽  
Yeast Saccharomyces Cerevisiae ◽  
Tolerant Strain ◽  
Cell Factories

Saccharomyces cerevisiae is an important unicellular yeast species within the biotechnological and food and beverage industries. A significant application of this species is the production of ethanol, where concentrations are limited by cellular toxicity, often at the level of the cell membrane. Here, we characterize 61 S. cerevisiae strains for ethanol tolerance and further analyse five representatives with varying ethanol tolerances. The most tolerant strain, AJ4, was dominant in co-culture at 0% and 10% ethanol. Unexpectedly, although it does not have the highest NIC or MIC, MY29 was the dominant strain in co-culture at 6% ethanol, which may be linked to differences in its basal lipidome. Whilst relatively few lipidomic differences were observed between strains, a significantly higher PE concentration was observed in the least tolerant strain, MY26, at 0% and 6% ethanol compared to the other strains that became more similar at 10%, indicating potential involvement of this lipid with ethanol sensitivity. Our findings reveal that AJ4 is best able to adapt its membrane to become more fluid in the presence of ethanol and lipid extracts from AJ4 also form the most permeable membranes. Furthermore, MY26 is least able to modulate fluidity in response to ethanol and membranes formed from extracted lipids are least leaky at physiological ethanol concentrations. Overall, these results reveal a potential mechanism of ethanol tolerance and suggests a limited set of membrane compositions that diverse yeast species use to achieve this. Importance Many microbial processes are not implemented at the industrial level because the product yield is poorer and more expensive than can be achieved by chemical synthesis. It is well established that microbes show stress responses during bioprocessing, and one reason for poor product output from cell factories is production conditions that are ultimately toxic to the cells. During fermentative processes, yeast cells encounter culture media with high sugar content, which is later transformed into high ethanol concentrations. Thus, ethanol toxicity is one of the major stresses in traditional and more recent biotechnological processes. We have performed a multilayer phenotypic and lipidomic characterization of a large number of industrial and environmental strains of Saccharomyces to identify key resistant and non-resistant isolates for future applications.

scholarly journals Harnessing the yeast Saccharomyces cerevisiae for the production of fungal secondary metabolites

2021 ◽  
Author(s):  
Guokun Wang ◽  
Douglas B. Kell ◽  
Irina Borodina
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Secondary Metabolites ◽  
High Throughput Screening ◽  
Large Scale ◽  
Strain Improvement ◽  
Cell Factory ◽  
Yeast Saccharomyces Cerevisiae ◽  
Cell Factories ◽  
Genetically Engineer ◽  
Fungal Secondary Metabolites

Abstract Fungal secondary metabolites (FSMs) represent a remarkable array of bioactive compounds, with potential applications as pharmaceuticals, nutraceuticals, and agrochemicals. However, these molecules are typically produced only in limited amounts by their native hosts. The native organisms may also be difficult to cultivate and genetically engineer, and some can produce undesirable toxic side-products. Alternatively, recombinant production of fungal bioactives can be engineered into industrial cell factories, such as aspergilli or yeasts, which are well amenable for large-scale manufacturing in submerged fermentations. In this review, we summarize the development of baker’s yeast Saccharomyces cerevisiae to produce compounds derived from filamentous fungi and mushrooms. These compounds mainly include polyketides, terpenoids, and amino acid derivatives. We also describe how native biosynthetic pathways can be combined or expanded to produce novel derivatives and new-to-nature compounds. We describe some new approaches for cell factory engineering, such as genome-scale engineering, biosensor-based high-throughput screening, and machine learning, and how these tools have been applied for S. cerevisiae strain improvement. Finally, we prospect the challenges and solutions in further development of yeast cell factories to more efficiently produce FSMs.

scholarly journals The Model System Saccharomyces cerevisiae Versus Emerging Non-Model Yeasts for the Production of Biofuels

Life ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 299
Author(s):  
Maria Priscila Lacerda ◽  
Eun Joong Oh ◽  
Carrie Eckert
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Metabolic Engineering ◽  
Synthetic Biology ◽  
Biofuel Cell ◽  
Efficient Production ◽  
Yeast Saccharomyces Cerevisiae ◽  
Fatty Acid Production ◽  
Inhibitor Tolerance ◽  
Cell Factories ◽  
Recent Advances

Microorganisms are effective platforms for the production of a variety of chemicals including biofuels, commodity chemicals, polymers and other natural products. However, deep cellular understanding is required for improvement of current biofuel cell factories to truly transform the Bioeconomy. Modifications in microbial metabolic pathways and increased resistance to various types of stress caused by the production of these chemicals are crucial in the generation of robust and efficient production hosts. Recent advances in systems and synthetic biology provide new tools for metabolic engineering to design strategies and construct optimal biocatalysts for the sustainable production of desired chemicals, especially in the case of ethanol and fatty acid production. Yeast is an efficient producer of bioethanol and most of the available synthetic biology tools have been developed for the industrial yeast Saccharomyces cerevisiae. Non-conventional yeast systems have several advantageous characteristics that are not easily engineered such as ethanol tolerance, low pH tolerance, thermotolerance, inhibitor tolerance, genetic diversity and so forth. Currently, synthetic biology is still in its initial steps for studies in non-conventional yeasts such as Yarrowia lipolytica, Kluyveromyces marxianus, Issatchenkia orientalis and Pichia pastoris. Therefore, the development and application of advanced synthetic engineering tools must also focus on these underexploited, non-conventional yeast species. Herein, we review the basic synthetic biology tools that can be applied to the standard S. cerevisiae model strain, as well as those that have been developed for non-conventional yeasts. In addition, we will discuss the recent advances employed to develop non-conventional yeast strains that are efficient for the production of a variety of chemicals through the use of metabolic engineering and synthetic biology.

scholarly journals Modulation of the voltage-dependent anion-selective channel by cytoplasmic proteins from wild type and the channel depleted cells of Saccharomyces cerevisiae.

2003 ◽  
Vol 50 (2) ◽  
pp. 415-424 ◽  
Author(s):  
Hanna Kmita ◽  
Małgorzata Budzińska ◽  
Olgierd Stobienia
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein S ◽  
Mitochondrial Outer Membrane ◽  
Cell Physiology ◽  
Intermembrane Space ◽  
Wild Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
Cytoplasmic Proteins ◽  
Voltage Dependent ◽  
Selective Channel

It is well known that effective exchange of metabolites between mitochondria and the cytoplasm is essential for cell physiology. The key step of the exchange is transport across the mitochondrial outer membrane, which is supported by the voltage-dependent anion-selective channel (VDAC). Therefore, it is clear that the permeability of VDAC must be regulated to adjust its activity to the actual cell needs. VDAC-modulating activities, often referred to as the VDAC modulator, were identified in the intermembrane space of different organism mitochondria but the responsible protein(s) has not been identified as yet. Because the VDAC modulator was reported to act on VDAC of intact mitochondria when added to the cytoplasmic side it has been speculated that a similar modulating activity might be present in the cytoplasm. To check the speculation we used mitochondria of the yeast Saccharomyces cerevisiae as they constitute a perfect model to study VDAC modulation. The mitochondria contain only a single isoform of VDAC and it is possible to obtain viable mutants devoid of the channel (Deltapor1). Moreover, we have recently characterised a VDAC-modulating activity located in the intermembrane space of wild type and Deltapor1 S. cerevisiae mitochondria. Here, we report that the cytoplasm of wild type and Deltapor1 cells of S. cerevisiae contains a VDAC-modulating activity as measured in a reconstituted system and with intact mitochondria. Since quantitative differences were observed between the modulating fractions isolated from wild type and Deltapor1 cells when they were studied with intact wild type mitochondria as well as by protein electrophoresis it might be concluded that VDAC may influence the properties of the involved cytoplasmic proteins. Moreover, the VDAC-modulating activity in the cytoplasm differs distinctly from that reported for the mitochondrial intermembrane space. Nevertheless, both these activities may contribute efficiently to VDAC regulation. Thus, the identification of the proteins is very important.

scholarly journals The influence of depletion of voltage dependent anion selective channel on protein import into the yeast Saccharomyces cerevisiae mitochondria.

2001 ◽  
Vol 48 (3) ◽  
pp. 719-728 ◽  
Author(s):  
A Szczechowicz ◽  
L Hryniewiecka ◽  
H Kmita
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fusion Protein ◽  
Outer Membrane ◽  
Protein Import ◽  
Wild Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
Tom Complex ◽  
Saccharomyces Cerevisiae Mitochondria ◽  
Voltage Dependent ◽  
Selective Channel

The supply of substrates to the respiratory chain as well as of other metabolites (e.g. ATP) into inner compartments of mitochondria is crucial to preprotein import into these organelles. Transport of the compounds across the outer mitochondrial membrane is enabled by mitochondrial porin, also known as the voltage-dependent anion-selective channel (VDAC). Our previous studies led to the conclusion that the transport of metabolites through the outer membrane of the yeast Saccharomyces cerevisiae mitochondria missing VDAC (now termed YVDAC1) is considerably restricted. Therefore we expected that depletion of YVDAC1 should also hamper protein import into the mutant mitochondria. We report here that YVDAC1-depleted mitochondria are able to import a fusion protein termed pSu9-DHFR in the amount comparable to that of wild type mitochondria, although over a considerably longer time. The rate of import of the fusion protein into YVDAC1-depleted mitochondria is dis- tinctly lower than into wild type mitochondria probably due to restricted ATP access to the intermembrane space and is additionally influenced by the way the supporting respiratory substrates are transported through the outer membrane. In the presence of ethanol, diffusing freely through lipid membranes, YVDAC1-depleted mitochondria are able to import the fusion protein at a higher rate than in the presence of external NADH which is, like ATP, transported through the outer membrane by facilitated diffusion. It has been shown that transport of external NADH across the outer membrane of YVDAC1-depleted mitochondria is supported by the protein import machinery, i.e. the TOM complex (Kmita & Budzińska, 2000, Biochim. Biophys. Acta 1509, 86-94.). Since the TOM complex might also contribute to the permeability of the membrane to ATP, it seems possible that external NADH and ATP as well as the imported preprotein could compete with one another for the passage through the outer membrane in YVDAC1-depleted mitochondria.

Sign in / Sign up

Export Citation Format

Share Document