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Engineering Saccharomyces cerevisiae for fast vitamin-independent aerobic growth

Ehrmann, Anja K. (author), Wronska, A.K. (author), Perli, T. (author), de Hulster, A.F. (author), Luttik, M.A.H. (author), van den Broek, M.A. (author), Carqueija Cardoso, D.C. (author), Pronk, J.T. (author), Daran, J.G. (author)

Chemically defined media for cultivation of Saccharomyces cerevisiae strains are commonly supplemented with a mixture of multiple Class-B vitamins, whose omission leads to strongly reduced growth rates. Fast growth without vitamin supplementation is interesting for industrial applications, as it reduces costs and complexity of medium...

journal article 2024

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Draft genome sequence of the Saccharomyces cerevisiae SpyCas9 expressing strain IMX2600, a laboratory and platform strain from the CEN.PK lineage for cell-factory research

van den Broek, M.A. (author), Ortiz Merino, R.A. (author), Bennis, N.X. (author), Wronska, A.K. (author), Else-Hassing, J. (author), Daran-Lapujade, P.A.S. (author), Daran, J.G. (author)

The biobased-economy aims to create a circular biotechnology ecosystem to transition from a fossil fuel-based to a sustainable industry based on biomass. For this, new microbial cell-factories are essential. We present the draft genome of the CEN.PK-derived Saccharomyces cerevisiae SpyCas9 expressing strain (IMX2600), that serve as chassis of...

journal article 2024

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CRI-SPA: a high-throughput method for systematic genetic editing of yeast libraries

Cachera, Paul (author), Olsson, Helén (author), Coumou, Hilde (author), Jensen, Mads L. (author), Sánchez, Benjamín J. (author), Strucko, Tomas (author), van den Broek, M.A. (author), Daran, J.G. (author), Jensen, Michael K. (author), Sonnenschein, Nikolaus (author), Lisby, Michael (author), Mortensen, Uffe H. (author)

Biological functions are orchestrated by intricate networks of interacting genetic elements. Predicting the interaction landscape remains a challenge for systems biology and new research tools allowing simple and rapid mapping of sequence to function are desirable. Here, we describe CRI-SPA, a method allowing the transfer of chromosomal...

journal article 2023

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Improving CRISPR-Cas9 mediated genome integration in interspecific hybrid yeasts

Bennis, N.X. (author), Kostanjsek, M. (author), van den Broek, M.A. (author), Daran, J.G. (author)

Saccharomyces pastorianus is not a classical taxon, it is an interspecific hybrid resulting from the cross of Saccharomyces cerevisiae and Saccharomyces eubayanus. Exhibiting heterosis for phenotypic traits such as wort α-oligosaccharide consumption and fermentation at low temperature, it has been domesticated to become the main workhorse of...

journal article 2023

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Long-read direct RNA sequencing of the mitochondrial transcriptome of Saccharomyces cerevisiae reveals condition-dependent intron abundance

Koster, C.C. (author), Kleefeldt, Askar A. (author), van den Broek, M.A. (author), Luttik, M.A.H. (author), Daran, J.G. (author), Daran-Lapujade, P.A.S. (author)

Mitochondria fulfil many essential roles and have their own genome, which is expressed as polycistronic transcripts that undergo co- or posttranscriptional processing and splicing. Due to the inherent complexity and limited technical accessibility of the mitochondrial transcriptome, fundamental questions regarding mitochondrial gene...

journal article 2023

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Modular, synthetic chromosomes as new tools for large scale engineering of metabolism

Postma, E.D. (author), Else-Hassing, J. (author), Mangkusaputra, Venda (author), Geelhoed, J. (author), de la Torre, P. (author), van den Broek, M.A. (author), Mooiman, C. (author), Pabst, Martin (author), Daran, J.G. (author), Daran-Lapujade, P.A.S. (author)

The construction of powerful cell factories requires intensive genetic engineering for the addition of new functionalities and the remodeling of native pathways and processes. The present study demonstrates the feasibility of extensive genome reprogramming using modular, specialized de novo-assembled neochromosomes in yeast. The in vivo...

journal article 2022

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A supernumerary designer chromosome for modular in vivo pathway assembly in Saccharomyces cerevisiae

Postma, E.D. (author), Dashko, S. (author), van Breemen, Lars (author), Taylor Parkins, S.K. (author), van den Broek, M.A. (author), Daran, J.G. (author), Daran-Lapujade, P.A.S. (author)

The construction of microbial cell factories for sustainable production of chemicals and pharmaceuticals requires extensive genome engineering. Using Saccharomyces cerevisiae, this study proposes synthetic neochromosomes as orthogonal expression platforms for rewiring native cellular processes and implementing new functionalities....

journal article 2021

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Engineering oxygen-independent biotin biosynthesis in Saccharomyces cerevisiae

Wronska, A.K. (author), van den Broek, M.A. (author), Perli, T. (author), de Hulster, A.F. (author), Pronk, J.T. (author), Daran, J.G. (author)

An oxygen requirement for de novo biotin synthesis in Saccharomyces cerevisiae precludes the application of biotin-prototrophic strains in anoxic processes that use biotin-free media. To overcome this issue, this study explores introduction of the oxygen-independent Escherichia coli biotin-biosynthesis pathway in S. cerevisiae. Implementation...

journal article 2021

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Engineering heterologous molybdenum-cofactor-biosynthesis and nitrate-assimilation pathways enables nitrate utilization by Saccharomyces cerevisiae

Perli, T. (author), van der Vorm, Daan N.A. (author), Wassink, Mats (author), van den Broek, M.A. (author), Pronk, J.T. (author), Daran, J.G. (author)

Metabolic capabilities of cells are not only defined by their repertoire of enzymes and metabolites, but also by availability of enzyme cofactors. The molybdenum cofactor (Moco) is widespread among eukaryotes but absent from the industrial yeast Saccharomyces cerevisiae. No less than 50 Moco-dependent enzymes covering over 30 catalytic...

journal article 2021

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Design and Experimental Evaluation of a Minimal, Innocuous Watermarking Strategy to Distinguish Near-Identical DNA and RNA Sequences

Boonekamp, F.J. (author), Dashko, S. (author), Duiker, Donna (author), Gehrmann, T. (author), van den Broek, M.A. (author), den Ridder, M.J. (author), Pabst, Martin (author), Robert, Vincent (author), Abeel, T.E.P.M.F. (author), Postma, E.D. (author), Daran, J.G. (author), Daran-Lapujade, P.A.S. (author)

The construction of powerful cell factories requires intensive and extensive remodelling of microbial genomes. Considering the rapidly increasing number of these synthetic biology endeavors, there is an increasing need for DNA watermarking strategies that enable the discrimination between synthetic and native gene copies. While it is well...

journal article 2020

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Improving Industrially Relevant Phenotypic Traits by Engineering Chromosome Copy Number in Saccharomyces pastorianus

Gorter de Vries, A.R. (author), Knibbe, E. (author), van Roosmalen, Roderick (author), van den Broek, M.A. (author), de la Torre, P. (author), O’Herne, Stephanie F. (author), Vijverberg, Pascal A. (author), el Masoudi, Anissa (author), Brouwers, N. (author), Pronk, J.T. (author), Daran, J.G. (author)

The lager-brewing yeast Saccharomyces pastorianus is a hybrid between S. cerevisiae and S. eubayanus with an exceptional degree of aneuploidy. While chromosome copy number variation (CCNV) is present in many industrial Saccharomyces strains and has been linked to various industrially-relevant traits, its impact on the brewing performance of S...

journal article 2020

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Exploiting the Diversity of Saccharomycotina Yeasts To Engineer Biotin-Independent Growth of Saccharomyces cerevisiae

Wronska, A.K. (author), Haak, Meinske P. (author), Geraats, Ellen (author), Bruins Slot, Eva (author), van den Broek, M.A. (author), Pronk, J.T. (author), Daran, J.G. (author)

Biotin, an important cofactor for carboxylases, is essential for all kingdoms of life. Since native biotin synthesis does not always suffice for fast growth and product formation, microbial cultivation in research and industry often requires supplementation of biotin. De novo biotin biosynthesis in yeasts is not fully understood, which...

journal article 2020

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Adaptive Laboratory Evolution and Reverse Engineering of Single-Vitamin Prototrophies in Saccharomyces cerevisiae

Perli, T. (author), Moonen, Dewi P.I. (author), van den Broek, M.A. (author), Pronk, J.T. (author), Daran, J.G. (author)

Quantitative physiological studies on Saccharomyces cerevisiae commonly use synthetic media (SM) that contain a set of water-soluble growth factors that, based on their roles in human nutrition, are referred to as B vitamins. Previous work demonstrated that in S. cerevisiae CEN. PK113-7D, requirements for biotin were eliminated by laboratory...

journal article 2020

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Exploring the abundance of oleate hydratases in the genus Rhodococcus—discovery of novel enzymes with complementary substrate scope

Busch, H. (author), Tonin, F. (author), Alvarenga Da Silva, N. (author), van den Broek, M.A. (author), Lu, Simona (author), Daran, J.G. (author), Hanefeld, U. (author), Hagedoorn, P.L. (author)

Oleate hydratases (Ohys, EC 4.2.1.53) are a class of enzymes capable of selective water addition reactions to a broad range of unsaturated fatty acids leading to the respective chiral alcohols. Much research was dedicated to improving the applications of existing Ohys as well as to the identification of undescribed Ohys with potentially novel...

journal article 2020

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The complete genome sequence of the nitrile biocatalyst Rhodocccus rhodochrous ATCC BAA-870

Frederick, Joni (author), Hennessy, Fritha (author), Horn, Uli (author), de la Torre Cortés, Pilar (author), van den Broek, M.A. (author), Strych, Ulrich (author), Willson, Richard (author), Daran, J.G. (author), Otten, L.G. (author)

BACKGROUND: Rhodococci are industrially important soil-dwelling Gram-positive bacteria that are well known for both nitrile hydrolysis and oxidative metabolism of aromatics. Rhodococcus rhodochrous ATCC BAA-870 is capable of metabolising a wide range of aliphatic and aromatic nitriles and amides. The genome of the organism was sequenced and...

journal article 2020

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In vivo recombination of Saccharomyces eubayanus maltose-transporter genes yields a chimeric transporter that enables maltotriose fermentation

Brouwers, N. (author), Gorter de Vries, A.R. (author), van den Broek, M.A. (author), Weening, S.M. (author), Elink Schuurman, Tom D. (author), Kuijpers, Niels G.A. (author), Pronk, J.T. (author), Daran, J.G. (author)

Saccharomyces eubayanus is the non-S. cerevisiae parent of the lager-brewing hybrid S. pastorianus. In contrast to most S. cerevisiae and Frohberg-type S. pastorianus strains, S. eubayanus cannot utilize the α-tri-glucoside maltotriose, a major carbohydrate in brewer's wort. In Saccharomyces yeasts, utilization of maltotriose is encoded by...

journal article 2019

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Chromosome level assembly and comparative genome analysis confirm lager-brewing yeasts originated from a single hybridization

Salazar, A.N. (author), Gorter de Vries, A.R. (author), van den Broek, M.A. (author), Brouwers, N. (author), de la Torre, P. (author), Kuijpers, N.G.A. (author), Daran, J.G. (author), Abeel, T.E.P.M.F. (author)

Background: The lager brewing yeast, S. pastorianus, is a hybrid between S. cerevisiae and S. eubayanus with extensive chromosome aneuploidy. S. pastorianus is subdivided into Group 1 and Group 2 strains, where Group 2 strains have higher copy number and a larger degree of heterozygosity for S. cerevisiae chromosomes. As a result, Group 2...

journal article 2019

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Laboratory Evolution of a Saccharomyces cerevisiae × S. eubayanus Hybrid Under Simulated Lager-Brewing Conditions

Gorter de Vries, A.R. (author), Voskamp, Maaike (author), van Aalst, Aafke (author), Kristensen, L.H. (author), Jansen, Liset (author), Salazar, A.N. (author), van den Broek, M.A. (author), Brouwers, N. (author), Abeel, T.E.P.M.F. (author), Pronk, J.T. (author), Daran, J.G. (author)

Saccharomyces pastorianus lager-brewing yeasts are domesticated hybrids of S. cerevisiae x S. eubayanus that display extensive inter-strain chromosome copy number variation and chromosomal recombinations. It is unclear to what extent such genome rearrangements are intrinsic to the domestication of hybrid brewing yeasts and whether they...

journal article 2019

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The Genetic Makeup and Expression of the Glycolytic and Fermentative Pathways Are Highly Conserved Within the Saccharomyces Genus

Boonekamp, F.J. (author), Dashko, S. (author), van den Broek, M.A. (author), Gehrmann, T. (author), Daran, J.G. (author), Daran-Lapujade, P.A.S. (author)

The ability of the yeast Saccharomyces cerevisiae to convert glucose, even in the presence of oxygen, via glycolysis and the fermentative pathway to ethanol has played an important role in its domestication. Despite the extensive knowledge on these pathways in S. cerevisiae, relatively little is known about their genetic makeup in other...

journal article 2018

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Structural, physiological and regulatory analysis of maltose transporter genes in Saccharomyces eubayanus CBS 12357^T

Brickwedde, A. (author), Brouwers, N. (author), van den Broek, M.A. (author), Gallego Murillo, Joan Sebastián (author), Fraiture, J.L. (author), Pronk, J.T. (author), Daran, J.G. (author)

Saccharomyces pastorianus lager brewing yeasts are domesticated hybrids of Saccharomyces cerevisiae and cold-tolerant Saccharomyces eubayanus. To understand the contribution of both parental genomes to maltose metabolism in brewing wort, this study focuses on maltose transport in the S. eubayanus type strain CBS 12357^T/FM1318. To...

journal article 2018