document

amdSYM, a new dominant recyclable marker cassette for Saccharomyces cerevisiae

Solis-Escalante, D. (author), Kuijpers, N.G.A. (author), Bongaerts, N. (author), Bolat, I. (author), Bosman, L. (author), Pronk, J.T. (author), Daran, J.M. (author), Daran-Lapujade, P.A.S. (author)

Despite the large collection of selectable marker genes available for Saccharomyces cerevisiae, marker availability can still present a hurdle when dozens of genetic manipulations are required. Recyclable markers, counterselectable cassettes that can be removed from the targeted genome after use, are therefore valuable assets in ambitious...

journal article 2012

document

Carbon dioxide fixation by Calvin-Cycle enzymes improves ethanol yield in yeast

Guadalupe-Medina, V. (author), Wisselink, H.W. (author), Luttik, M.A.H. (author), De Hulster, E. (author), Daran, J.M. (author), Pronk, J.T. (author), Van Maris, A.J.A. (author)

Background Redox-cofactor balancing constrains product yields in anaerobic fermentation processes. This challenge is exemplified by the formation of glycerol as major by-product in yeast-based bioethanol production, which is a direct consequence of the need to reoxidize excess NADH and causes a loss of conversion efficiency. Enabling the use of...

journal article 2013

document

Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae

Milne, N. (author), Luttik, M.A.H. (author), Cueto Rojas, H.F. (author), Wahl, A. (author), Van Maris, A.J.A. (author), Pronk, J.T. (author), Daran, J.G. (author)

In microbial processes for production of proteins, biomass and nitrogen-containing commodity chemicals, ATP requirements for nitrogen assimilation affect product yields on the energy producing substrate. In Saccharomyces cerevisiae, a current host for heterologous protein production and potential platform for production of nitrogen-containing...

journal article 2015

document

Alternative reactions at the interface of glycolysis and citric acid cycle in Saccharomyces cerevisiae

Rossum, Harmen M. (author), Kozak, B.U. (author), Niemeijer, M.S. (author), Duine, Hendrik J. (author), Luttik, M.A.H. (author), Boer, VM (author), Kötter, P (author), Daran, J.G. (author), van Maris, A.J.A. (author), Pronk, J.T. (author)

Pyruvate and acetyl-coenzyme A, located at the interface between glycolysis and TCA cycle, are important intermediates in yeast metabolism and key precursors for industrially relevant products. Rational engineering of their supply requires knowledge of compensatory reactions that replace predominant pathways when these are inactivated. This...

journal article 2016

document

Nanopore sequencing enables near-complete de novo assembly of Saccharomyces cerevisiae reference strain CEN.PK113-7D

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

document

Laboratory evolution of a biotin-requiring Saccharomyces cerevisiae strain for full biotin prototrophy and identification of causal mutations

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

document

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

document

Engineering de novo anthocyanin production in Saccharomyces cerevisiae

Levisson, Mark (author), Patinios, Constantinos (author), Hein, Sascha (author), de Groot, P.A. (author), Daran, J.G. (author), Martens, Stefan (author), Hall, Robert D. (author), Beekwilder, Jules (author)

Background: Anthocyanins are polyphenolic pigments which provide pink to blue colours in fruits and flowers. There is an increasing demand for anthocyanins, as food colorants and as health-promoting substances. Plant production of anthocyanins is often seasonal and cannot always meet demand due to low productivity and the complexity of the...

journal article 2018

document

Toward developing a yeast cell factory for the production of prenylated flavonoids

Levisson, Mark (author), Araya-Cloutier, Carla (author), De Bruijn, Wouter J.C. (author), Van Der Heide, Menno (author), Salvador López, José Manuel (author), Daran, J.G. (author), Vincken, Jean Paul (author), Beekwilder, Jules (author)

Prenylated flavonoids possess a wide variety of biological activities, including estrogenic, antioxidant, antimicrobial, and anticancer activities. Hence, they have potential applications in food products, medicines, or supplements with health-promoting activities. However, the low abundance of prenylated flavonoids in nature is limiting...

journal article 2019

document

Connecting central carbon and aromatic amino acid metabolisms to improve de novo 2-phenylethanol production in Saccharomyces cerevisiae

Else-Hassing, J. (author), de Groot, P.A. (author), Marquenie, Vita R. (author), Pronk, J.T. (author), Daran, J.G. (author)

The organic compound 2-phenylethanol (2PE) has a pleasant floral scent and is intensively used in the cosmetic and food industries. Microbial production of 2PE by phenylalanine bioconversion or de novo biosynthesis from sugar offer sustainable, reliable and natural production processes compared to chemical synthesis. Despite the ability of...

journal article 2019

document

Vitamin requirements and biosynthesis in Saccharomyces cerevisiae

Perli, T. (author), Wronska, A.K. (author), Ortiz Merino, R.A. (author), Pronk, J.T. (author), Daran, J.G. (author)

Chemically defined media for yeast cultivation (CDMY) were developed to support fast growth, experimental reproducibility, and quantitative analysis of growth rates and biomass yields. In addition to mineral salts and a carbon substrate, popular CDMYs contain seven to nine B-group vitamins, which are either enzyme cofactors or precursors for...

journal article 2020

document

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

document

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

document

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

document

Identification of Oxygen-Independent Pathways for Pyridine Nucleotide and Coenzyme A Synthesis in Anaerobic Fungi by Expression of Candidate Genes in Yeast

Perli, T. (author), Vos, A.M. (author), Bouwknegt, J. (author), Dekker, W.J.C. (author), Wiersma, S.J. (author), Mooiman, C. (author), Ortiz Merino, R.A. (author), Daran, J.G. (author), Pronk, J.T. (author)

Neocallimastigomycetes are unique examples of strictly anaerobic eukaryotes. This study investigates how these anaerobic fungi bypass reactions involved in synthesis of pyridine nucleotide cofactors and coenzyme A that, in canonical fungal pathways, require molecular oxygen. Analysis of Neocallimastigomycetes proteomes identified a candidate...

journal article 2021

document

Elimination of aromatic fusel alcohols as by-products of Saccharomyces cerevisiae strains engineered for phenylpropanoid production by 2-oxo-acid decarboxylase replacement

Else-Hassing, J. (author), Buijs, Joran (author), Blankerts, Nikki (author), Luttik, M.A.H. (author), de Hulster, A.F. (author), Pronk, J.T. (author), Daran, J.G. (author)

Engineered strains of the yeast Saccharomyces cerevisiae are intensively studied as production platforms for aromatic compounds such as hydroxycinnamic acids, stilbenoids and flavonoids. Heterologous pathways for production of these compounds use L-phenylalanine and/or L-tyrosine, generated by the yeast shikimate pathway, as aromatic...

journal article 2021

document

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

document

Full humanization of the glycolytic pathway in Saccharomyces cerevisiae

Boonekamp, F.J. (author), Knibbe, E. (author), Vieira Lara, M.A. (author), Wijsman, M. (author), Luttik, M.A.H. (author), van Eunen, Karen (author), den Ridder, M.J. (author), Pabst, Martin (author), Daran, J.G. (author), Daran-Lapujade, P.A.S. (author)

Although transplantation of single genes in yeast plays a key role in elucidating gene functionality in metazoans, technical challenges hamper humanization of full pathways and processes. Empowered by advances in synthetic biology, this study demonstrates the feasibility and implementation of full humanization of glycolysis in yeast. Single...

journal article 2022

document

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

document

Expanding the genome editing toolbox of Saccharomyces cerevisiae with the endonuclease ErCas12a

Bennis, N.X. (author), Anderson, Jonah P. (author), Kok, Siebe M.C. (author), Daran, J.G. (author)

ErCas12a is a class 2 type V CRISPR-Cas nuclease isolated from Eubacterium rectale with attractive fundamental characteristics, such as RNA self-processing capability, and lacks reach-through royalties typical for Cas nucleases. This study aims to develop a ErCas12a-mediated genome editing tool applicable in the model yeast Saccharomyces...

journal article 2023