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van Aalst, Aafke C.A. (author), Geraats, Ellen H. (author), Martini, M.L.A. (author), Mans, R. (author), Pronk, J.T. (author)In anaerobic Saccharomyces cerevisiae cultures, NADH (reduced form of nicotinamide adenine dinucleotide)-cofactor balancing by glycerol formation constrains ethanol yields. Introduction of an acetate-to-ethanol reduction pathway based on heterologous acetylating acetaldehyde dehydrogenase (A-ALD) can replace glycerol formation as 'redox-sink'...journal article 2023
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van Aalst, A.C.A. (author), van der Meulen, Igor S. (author), Jansen, Mickel L.A. (author), Mans, R. (author), Pronk, J.T. (author)Glycerol is the major organic byproduct of industrial ethanol production with the yeast Saccharomyces cerevisiae. Improved ethanol yields have been achieved with engineered S. cerevisiae strains in which heterologous pathways replace glycerol formation as the predominant mechanism for anaerobic re-oxidation of surplus NADH generated in...journal article 2023
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van Aalst, A.C.A. (author), Jansen, Mickel L.A. (author), Mans, R. (author), Pronk, J.T. (author)Background: Anaerobic Saccharomyces cerevisiae cultures require glycerol formation to re-oxidize NADH formed in biosynthetic processes. Introduction of the Calvin-cycle enzymes phosphoribulokinase (PRK) and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) has been shown to couple re-oxidation of biosynthetic NADH to ethanol...journal article 2023
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de Valk, S.C. (author), Mans, R. (author)A major challenge in the research of transport proteins is to understand how single amino acid residues contribute to their structure and biological function. Amino acid substitutions that result in a selective advantage in adaptive laboratory evolution experiments can provide valuable hints at their role in transport proteins. In this study, we...journal article 2022
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van Aalst, A.C.A. (author), de Valk, S.C. (author), van Gulik, W.M. (author), Jansen, Mickel L.A. (author), Pronk, J.T. (author), Mans, R. (author)Product yield on carbohydrate feedstocks is a key performance indicator for industrial ethanol production with the yeast Saccharomyces cerevisiae. This paper reviews pathway engineering strategies for improving ethanol yield on glucose and/or sucrose in anaerobic cultures of this yeast by altering the ratio of ethanol production, yeast growth...journal article 2022
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Malubhoy, Zahabiya (author), Bahia, Frederico Mendonça (author), de Valk, Sophie Claire (author), de Hulster, A.F. (author), Rendulić, Toni (author), Ortiz, Juan Paulo Ragas (author), Xiberras, Joeline (author), Klein, Mathias (author), Mans, R. (author), Nevoigt, Elke (author)Background: The microbial production of succinic acid (SA) from renewable carbon sources via the reverse TCA (rTCA) pathway is a process potentially accompanied by net-fixation of carbon dioxide (CO<sub>2</sub>). Among reduced carbon sources, glycerol is particularly attractive since it allows a nearly twofold higher CO<sub>2</sub>-fixation...journal article 2022
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de Hulster, A.F. (author), Mooiman, C. (author), Timmermans, Rowin (author), Mans, R. (author)Evolutionary engineering of microbes provides a powerful tool for untargeted optimization of (engineered) cell factories and identification of genetic targets for further research. Directed evolution is an intrinsically time-intensive effort, and automated methods can significantly reduce manual labor. Here, design considerations for various...journal article 2022
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de Valk, S.C. (author), Bouwmeester, Susan E. (author), de Hulster, A.F. (author), Mans, R. (author)Background: In the yeast Saccharomyces cerevisiae, which is widely applied for industrial bioethanol production, uptake of hexoses is mediated by transporters with a facilitated diffusion mechanism. In anaerobic cultures, a higher ethanol yield can be achieved when transport of hexoses is proton-coupled, because of the lower net ATP yield of...journal article 2022
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van Winden, Wouter A. (author), Mans, R. (author), Breestraat, Stefaan (author), Verlinden, Rob A.J. (author), Mielgo Gomez, A. (author), de Hulster, A.F. (author), de Bruijn, J.A. (author), Noorman, H.J. (author)A novel fermentation process was developed in which renewable electricity is indirectly used as an energy source in fermentation, synergistically decreasing both the consumption of sugar as a first generation carbon source and emission of the greenhouse gas CO<sub>2</sub>. As an illustration, a glucose-based process is co-fed with formic acid...journal article 2022
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van Aalst, A.C.A. (author), Mans, R. (author), Pronk, J.T. (author)Background: Saccharomyces cerevisiae is intensively used for industrial ethanol production. Its native fermentation pathway enables a maximum product yield of 2 mol of ethanol per mole of glucose. Based on conservation laws, supply of additional electrons could support even higher ethanol yields. However, this option is disallowed by the...journal article 2022
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Dekker, W.J.C. (author), Jürgens, H. (author), Ortiz Merino, R.A. (author), Mooiman, C. (author), van den Berg, Remon (author), Kaljouw, Astrid (author), Mans, R. (author), Pronk, J.T. (author)While thermotolerance is an attractive trait for yeasts used in industrial ethanol production, oxygen requirements of known thermotolerant species are incompatible with process requirements. Analysis of oxygen-sufficient and oxygen-limited chemostat cultures of the facultatively fermentative, thermotolerant species Ogataea parapolymorpha...journal article 2022
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Baldi, N. (author), de Valk, S.C. (author), Sousa-Silva, Maria (author), Casal, Margarida (author), Soares-Silva, Isabel (author), Mans, R. (author)In Saccharomyces cerevisiae, the complete set of proteins involved in transport of lactic acid across the cell membrane has not been determined. In this study, we aimed to identify transport proteins not previously described to be involved in lactic acid transport via a combination of directed evolution, whole-genome resequencing and reverse...journal article 2021
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Barata-Antunes, Cláudia (author), Alves, Rosana (author), Talaia, Gabriel (author), Casal, Margarida (author), Gerós, Hernâni (author), Mans, R. (author), Paiva, Sandra (author)Plasma membrane transporters play pivotal roles in the import of nutrients, including sugars, amino acids, nucleobases, carboxylic acids, and metal ions, that surround fungal cells. The selective removal of these transporters by endocytosis is one of the most important regulatory mechanisms that ensures a rapid adaptation of cells to the...review 2021
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D'Ambrosio, Vasil (author), Dore, Eleonora (author), Di Blasi, Roberto (author), van den Broek, M.A. (author), Sudarsan, Suresh (author), Horst, Jolanda ter (author), Ambri, Francesca (author), Keasling, Jay D. (author), Mans, R. (author)Engineering living cells for production of chemicals, enzymes and therapeutics can burden cells due to use of limited native co-factor availability and/or expression burdens, totalling a fitness deficit compared to parental cells encoded through long evolutionary trajectories to maximise fitness. Ultimately, this discrepancy puts a selective...journal article 2020
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Henderson, Ryan K. (author), de Valk, Sophie C. (author), Poolman, Bert (author), Mans, R. (author)Proton coupled transport of α-glucosides via Mal11 into Saccharomyces cerevisiae costs one ATP per imported molecule. Targeted mutation of all three acidic residues in the active site resulted in sugar uniport, but expression of these mutant transporters in yeast did not enable growth on sucrose. We then isolated six unique transporter...journal article 2020
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Xiberras, Joeline (author), Klein, Mathias (author), de Hulster, A.F. (author), Mans, R. (author), Nevoigt, Elke (author)Previously, our lab replaced the endogenous FAD-dependent pathway for glycerol catabolism in S. cerevisiae by the synthetic NAD-dependent dihydroxyacetone (DHA) pathway. The respective modifications allow the full exploitation of glycerol’s higher reducing power (compared to sugars) for the production of the platform chemical succinic acid ...journal article 2020
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Gabba, Matteo (author), Frallicciardi, Jacopo (author), van ’t Klooster, Joury (author), Henderson, Ryan (author), Syga, Łukasz (author), Mans, R. (author), van Maris, A.J.A. (author), Poolman, Bert (author)We present a fluorescence-based approach for determination of the permeability of small molecules across the membranes of lipid vesicles and living cells. With properly designed experiments, the method allows us to assess the membrane physical properties both in vitro and in vivo. We find that the permeability of weak acids increases in the...journal article 2020
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Baldi, N. (author), Dykstra, James C. (author), Luttik, M.A.H. (author), Pabst, Martin (author), Wu, Liang (author), Benjamin, Kirsten R. (author), Vente, André (author), Pronk, J.T. (author), Mans, R. (author)Efficient production of fuels and chemicals by metabolically engineered micro-organisms requires availability of precursor molecules for product pathways. In eukaryotic cell factories, heterologous product pathways are usually expressed in the cytosol, which may limit availability of precursors that are generated in other cellular...journal article 2019
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Mans, R. (author), Daran, J.G. (author), Pronk, J.T. (author)Evolutionary engineering, which uses laboratory evolution to select for industrially relevant traits, is a popular strategy in the development of high-performing yeast strains for industrial production of fuels and chemicals. By integrating whole-genome sequencing, bioinformatics, classical genetics and genome-editing techniques, evolutionary...review 2018
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Marques, W.L. (author), Mans, R. (author), Henderson, Ryan K. (author), Marella, Eko Roy (author), ter Horst, J. (author), de Hulster, A.F. (author), Poolman, Bert (author), Daran, J.G. (author), Pronk, J.T. (author), Gombert, Andreas K. (author), van Maris, A.J.A. (author)Anaerobic industrial fermentation processes do not require aeration and intensive mixing and the accompanying cost savings are beneficial for production of chemicals and fuels. However, the free-energy conservation of fermentative pathways is often insufficient for the production and export of the desired compounds and/or for cellular growth...journal article 2018
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