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Jürgens, H. (author), Varela, Javier A. (author), Gorter de Vries, A.R. (author), Perli, T. (author), Gast, V.J.M. (author), Gyurchev, N.Y. (author), Rajkumar, Arun S. (author), Mans, R. (author), Pronk, J.T. (author), Morrissey, John P. (author), Daran, J.G. (author)While CRISPR-Cas9-mediated genome editing has transformed yeast research, current plasmids and cassettes for Cas9 and guide-RNA expression are species specific. CRISPR tools that function in multiple yeast species could contribute to the intensifying research on non-conventional yeasts. A plasmid carrying a pangenomic origin of replication...journal article 2018
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Wijsman, M. (author), Swiat, M.A. (author), Marques, W.L. (author), Hettinga, Johanna K. (author), van den Broek, M.A. (author), Torre Cortés, Pilar de la (author), Mans, R. (author), Pronk, J.T. (author), Daran, J.G. (author), Daran-Lapujade, P.A.S. (author)Hexose transporter-deficient yeast strains are valuable testbeds for the study of sugar transport by native and heterologous transporters. In the popular Saccharomyces cerevisiae strain EBY.VW4000, deletion of 21 transporters completely abolished hexose transport. However, repeated use of the LoxP/Cre system in successive deletion rounds also...journal article 2018
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Verhoeven, M.D. (author), de Valk, S.C. (author), Daran, J.G. (author), van Maris, A.J.A. (author), Pronk, J.T. (author)D-Glucose, D-xylose and L-arabinose are major sugars in lignocellulosic hydrolysates. This study explores fermentation of glucose-xylose-arabinose mixtures by a consortium of three ‘specialist’ Saccharomyces cerevisiae strains. A D-glucose- and L-arabinose-tolerant xylose specialist was constructed by eliminating hexose phosphorylation in an...journal article 2018
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Verhoeven, M.D. (author), Bracher, J.M. (author), Nijland, Jeroen G. (author), Bouwknegt, J. (author), Daran, J.G. (author), Driessen, Arnold J.M. (author)Cas9-assisted genome editing was used to construct an engineered glucose-phosphorylation-negative S. cerevisiae strain, expressing the Lactobacillus plantaruml-arabinose pathway and the Penicillium chrysogenum transporter PcAraT. This strain, which showed a growth rate of 0.26 h−1 on l-arabinose in aerobic batch cultures, was subsequently...journal article 2018
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Diderich, J.A. (author), Weening, S.M. (author), van den Broek, M.A. (author), Pronk, J.T. (author), Daran, J.G. (author)Saccharomyces pastorianus is an interspecies hybrid between S. cerevisiae and S. eubayanus. The identification of the parental species of S. pastorianus enabled the de novo reconstruction of hybrids that could potentially combine a wide array of phenotypic traits. Lager yeasts are characterized by their inability to decarboxylate ferulic acid...journal article 2018
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Mans, R. (author), Wijsman, M. (author), Daran-Lapujade, P.A.S. (author), Daran, J.G. (author)Here, two methods are described for efficient genetic modification of Saccharomyces cerevisiae using CRISPR/Cas9. The first method enables the modification of a single genetic locus using in vivo assembly of a guide RNA (gRNA) expression plasmid without the need for prior cloning. A second method using in vitro assembled plasmids that could...journal article 2018
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Gorter de Vries, A.R. (author), Couwenberg, Lucas G.F. (author), van den Broek, M.A. (author), De La Torre Cortés, Pilar (author), ter Horst, J. (author), Pronk, J.T. (author), Daran, J.G. (author)Targeted DNA double-strand breaks (DSBs) with CRISPR-Cas9 have revolutionized genetic modification by enabling efficient genome editing in a broad range of eukaryotic systems. Accurate gene editing is possible with near-perfect efficiency in haploid or (predominantly) homozygous genomes. However, genomes exhibiting polyploidy and/or high...journal article 2018
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Mans, R. (author), Else-Hassing, J. (author), Wijsman, M. (author), Giezekamp, Annabel (author), Pronk, J.T. (author), Daran, J.G. (author), van Maris, A.J.A. (author)CRISPR/Cas9-based genome editing allows rapid, simultaneous modification of multiple genetic loci in Saccharomyces cerevisiae. Here, this technique was used in a functional analysis study aimed at identifying the hitherto unknown mechanism of lactate export in this yeast. First, an S. cerevisiae strain was constructed with deletions in 25...journal article 2017
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Swiat, M.A. (author), Dashko, S. (author), den Ridder, M.J. (author), Wijsman, M. (author), van der Oost, John (author), Daran, J.G. (author), Daran-Lapujade, P.A.S. (author)Cpf1 is a new class II family of CRISPR-Cas RNA-programmable endonucleases with unique features that make it a very attractive alternative or complement to Cas9 for genome engineering. Using constitutively expressed Cpf1 from Francisella novicida, the present study demonstrates that FnCpf1 can mediate RNA-guided DNA cleavage at targeted...journal article 2017
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Bracher, J.M. (author), de Hulster, A.F. (author), Koster, C.C. (author), van den Broek, M.A. (author), Daran, J.G. (author), van Maris, A.J.A. (author), Pronk, J.T. (author)Biotin prototrophy is a rare, incompletely understood, and industrially relevant characteristic of Saccharomyces cerevisiae strains. The genome of the haploid laboratory strain CEN.PK113-7D contains a full complement of biotin biosynthesis genes, but its growth in biotin-free synthetic medium is extremely slow (specific growth rate [μ] ≈ 0.01...journal article 2017
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Gorter de Vries, A.R. (author), Pronk, J.T. (author), Daran, J.G. (author)Chromosomal copy number variation (CCNV) plays a key role in evolution and health of eukaryotes. The unicellular yeast Saccharomyces cerevisiae is an important model for studying the generation, physiological impact, and evolutionary significance of CCNV. Fundamental studies of this yeast have contributed to an extensive set of methods for...journal article 2017
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Cueto Rojas, H.F. (author), Milne, Nicholas (author), van Helmond, W. (author), Pieterse, M.M. (author), Maris, Antonius J A (author), Daran, J.G. (author), Wahl, S.A. (author)Background: Microbial production of nitrogen containing compounds requires a high uptake flux and assimilation of the N-source (commonly ammonium), which is generally coupled with ATP consumption and negatively influences the product yield. In the industrial workhorse Saccharomyces cerevisiae, ammonium (NH<sub>4</sub> <sup>+</sup>) uptake is...journal article 2017
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Verhoeven, M.D. (author), Lee, Misun (author), Kamoen, L. (author), van den Broek, M.A. (author), Janssen, Dick B. (author), Daran, J.G. (author), van Maris, A.J.A. (author), Pronk, J.T. (author)Combined overexpression of xylulokinase, pentose-phosphate-pathway enzymes and a heterologous xylose isomerase (XI) is required but insufficient for anaerobic growth of Saccharomyces cerevisiae on d-xylose. Single-step Cas9-assisted implementation of these modifications yielded a yeast strain expressing Piromyces XI that showed fast aerobic...journal article 2017
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Brickwedde, A. (author), van den Broek, M.A. (author), Geertman, Jan Maarten A. (author), Magalhães, Frederico (author), Kuijpers, Niels G.A. (author), Gibson, Brian (author), Pronk, J.T. (author), Daran, J.G. (author)The lager brewing yeast Saccharomyces pastorianus, an interspecies hybrid of S. eubayanus and S. cerevisiae, ferments maltotriose, maltose, sucrose, glucose and fructose in wort to ethanol and carbon dioxide. Complete and timely conversion ("attenuation") of maltotriose by industrial S. pastorianus strains is a key requirement for process...journal article 2017
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Marques, W.L. (author), Mans, R. (author), Marella, Eko Roy (author), Cordeiro, Rosa Lorizolla (author), van den Broek, M.A. (author), Daran, J.G. (author), Pronk, J.T. (author), Gombert, Andreas K. (author), van Maris, A.J.A. (author)Many relevant options to improve efficacy and kinetics of sucrose metabolism in Saccharomyces cerevisiae and, thereby, the economics of sucrose-based processes remain to be investigated. An essential first step is to identify all native sucrose-hydrolysing enzymes and sucrose transporters in this yeast, including those that can be activated...journal article 2017
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Gorter de Vries, A.R. (author), de Groot, P.A. (author), van den Broek, M.A. (author), Daran, J.G. (author)Background: The ease of use of CRISPR-Cas9 reprogramming, its high efficacy, and its multiplexing capabilities have brought this technology at the forefront of genome editing techniques. Saccharomyces pastorianus is an aneuploid interspecific hybrid of Saccharomyces cerevisiae and Saccharomyces eubayanus that has been domesticated for...journal article 2017
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Salazar, A.N. (author), Gorter de Vries, A.R. (author), van den Broek, M.A. (author), Wijsman, M. (author), de la Torre, P. (author), Brickwedde, A. (author), Brouwers, N. (author), Daran, J.G. (author), Abeel, T.E.P.M.F. (author)The haploid Saccharomyces cerevisiae strain CEN.PK113–7D is a popular model system for metabolic engineering and systems biology research. Current genome assemblies are based on short-read sequencing data scaffolded based on homology to strain S288C. However, these assemblies contain large sequence gaps, particularly in subtelomeric regions, and...journal article 2017
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Gonzalez Ramos, D. (author), Gorter de Vries, A.R. (author), Grijseels, Sietske S. (author), van Berkum, M.C. (author), Swinnen, Steve (author), van den Broek, M.A. (author), Nevoigt, Elke (author), Daran, J.G. (author), Pronk, J.T. (author), van Maris, A.J.A. (author)Background: Acetic acid, released during hydrolysis of lignocellulosic feedstocks for second generation bioethanol production, inhibits yeast growth and alcoholic fermentation. Yeast biomass generated in a propagation step that precedes ethanol production should therefore express a high and constitutive level of acetic acid tolerance before...journal article 2016
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Stribny, Jiri (author), Romagnoli, G. (author), Perez Torrado, R. (author), Daran, J.G. (author), Querol, Amparo (author)Background: The yeast amino acid catabolism plays an important role in flavour generation since higher alcohols and acetate esters, amino acid catabolism end products, are key components of overall flavour and aroma in fermented products. Comparative studies have shown that other Saccharomyces species, such as S. kudriavzevii, differ during...journal article 2016
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Kozak, B.U. (author), Rossum, Harmen M. (author), Niemeijer, M.S. (author), van Dijk, M. (author), Benjamin, Kirsten (author), Wu, Liang (author), Daran, J.G. (author), Pronk, J.T. (author), van Maris, A.J.A. (author)In Saccharomyces cerevisiae ethanol dissimilation is initiated by its oxidation and activation to cytosolic acetyl-CoA. The associated consumption of ATP strongly limits yields of biomass and acetyl-CoA-derived products. Here, we explore the implementation of an ATP-independent pathway for acetyl-CoA synthesis from ethanol that, in theory,...journal article 2016