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Respiratory reoxidation of NADH is a key contributor to high oxygen requirements of oxygen-limited cultures of Ogataea parapolymorpha

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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Identification of fungal dihydrouracil-oxidase genes by expression in Saccharomyces cerevisiae

Bouwknegt, J. (author), Vos, A.M. (author), Ortiz Merino, R.A. (author), van Cuylenburg, Daphne C. (author), Luttik, M.A.H. (author), Pronk, J.T. (author)

Analysis of predicted fungal proteomes revealed a large family of sequences that showed similarity to the Saccharomyces cerevisiae Class-I dihydroorotate dehydrogenase Ura1, which supports synthesis of pyrimidines under aerobic and anaerobic conditions. However, expression of codon-optimised representatives of this gene family, from the...

journal article 2022

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An engineered non-oxidative glycolytic bypass based on Calvin-cycle enzymes enables anaerobic co-fermentation of glucose and sorbitol by Saccharomyces cerevisiae

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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A Framework for Evaluating Performance of Large-Scale Nature-Based Solutions to Reduce Hydro-Meteorological Risks and Enhance Co-benefits

Ruangpan, L. (author), Vojinovic, Zoran (author)

Over recent decades, hydro-meteorological disasters appear to be becoming more intense and frequent. Nature-Based Solutions (NBS) have been introduced to address hydro-meteorological risks as they offer the possibility of working closely with nature. This provides solutions to adapt to future changes in climate and society, as well as to...

book chapter 2022

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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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Critical parameters and procedures for anaerobic cultivation of yeasts in bioreactors and anaerobic chambers

Mooiman, C. (author), Bouwknegt, J. (author), Dekker, W.J.C. (author), Wiersma, S.J. (author), Ortiz Merino, R.A. (author), de Hulster, A.F. (author), Pronk, J.T. (author)

All known facultatively fermentative yeasts require molecular oxygen for growth. Only in a small number of yeast species, these requirements can be circumvented by supplementation of known anaerobic growth factors such as nicotinate, sterols and unsaturated fatty acids. Biosynthetic oxygen requirements of yeasts are typically small and,...

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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Natronoglycomyces albus gen. Nov., sp. nov, a haloalkaliphilic actinobacterium from a soda solonchak soil

Sorokin, Dimitry Y. (author), Khijniak, Tatjana V. (author), Zakharycheva, Alicia P. (author), Elcheninov, Alexander G. (author), Hahnke, Richard L. (author), Boueva, Olga V. (author), Ariskina, Elena V. (author), Bunk, Boyke (author), Kublanov, Ilya V. (author), Evtushenko, Lyudmila I. (author)

A haloalkaliphilic hydrolytic actinobacterium, strain ACPA22^T, was enriched and isolated in pure culture from saline alkaline soil (soda solonchak) in northeastern Mongolia. The isolate was facultatively alkaliphilic, growing at pH 6.5–10.5 (optimum at 7.3–9.0) and highly salt-tolerant, tolerating up to 3 M total Na⁺ as...

journal article 2021

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Uncoupling growth and succinic acid production in an industrial Saccharomyces cerevisiae strain

Liu, Y. (author), Esen, Osman (author), Pronk, J.T. (author), van Gulik, W.M. (author)

This study explores the relation between biomass-specific succinic acid (SA) production rate and specific growth rate of an engineered industrial strain of Saccharomyces cerevisiae, with the aim to investigate the extent to which growth and product formation can be uncoupled. Ammonium-limited aerobic chemostat and retentostat cultures were...

journal article 2021

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Engineering the thermotolerant industrial yeast Kluyveromyces marxianus for anaerobic growth

Dekker, W.J.C. (author), Ortiz Merino, R.A. (author), Kaljouw, Astrid (author), Battjes, Julius (author), Wiering, Frank W. (author), Mooiman, C. (author), de la Torre, P. (author), Pronk, J.T. (author)

Current large-scale, anaerobic industrial processes for ethanol production from renewable carbohydrates predominantly rely on the mesophilic yeast Saccharomyces cerevisiae. Use of thermotolerant, facultatively fermentative yeasts such as Kluyveromyces marxianus could confer significant economic benefits. However, in contrast to S. cerevisiae,...

journal article 2021

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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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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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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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Squalene-Tetrahymanol Cyclase Expression Enables Sterol-Independent Growth of Saccharomyces cerevisiae

Wiersma, S.J. (author), Mooiman, C. (author), Giera, Martin (author), Pronk, J.T. (author)

Biosynthesis of sterols, which are considered essential components of virtually all eukaryotic membranes, requires molecular oxygen. Anaerobic growth of the yeast Saccharomyces cerevisiae therefore strictly depends on sterol supplementation of synthetic growth media. Neocallimastigomycota are a group of strictly anaerobic fungi which, instead...

journal article 2020

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Hydroxynitrile lyases covalently immobilized in continuous flow microreactors

van der Helm, M. (author), Bracco Garcia, M.P. (author), Busch, H. (author), Szymańska, Katarzyna (author), Jarzȩbski, Andrzej B. (author), Hanefeld, U. (author)

Enzymes are supreme catalysts when it comes to high enantiopurities and their immobilization will pave the way for continuous operation. In this context, we show the covalent immobilization of hydroxynitrile lyases HbHNL (from Hevea brasiliensis) and MeHNL (from Manihot esculenta) in a siliceous monolithic microreactor for continuous...

journal article 2019

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Phenotype-independent isolation of interspecies Saccharomyces hybrids by dual-dye fluorescent staining and fluorescence-activated cell sorting

Gorter de Vries, A.R. (author), Koster, C.C. (author), Weening, S.M. (author), Luttik, M.A.H. (author), Kuijpers, N.G.A. (author), Geertman, Jan Maarten A. (author), Pronk, J.T. (author), Daran, J.G. (author)

Interspecies hybrids of Saccharomyces species are found in a variety of industrial environments and often outperform their parental strains in industrial fermentation processes. Interspecies hybridization is therefore increasingly considered as an approach for improvement and diversification of yeast strains for industrial application....

journal article 2019

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Selection of Pof^- saccharomyces eubayanus variants for the construction of S. cerevisiae × S. eubayanus hybrids with reduced 4-Vinyl guaiacol formation

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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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, J.S. (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

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Laboratory evolution for forced glucose-xylose co-consumption enables identification of mutations that improve mixed-sugar fermentation by xylose-fermenting Saccharomyces cerevisiae

Papapetridis, I. (author), Verhoeven, M.D. (author), Wiersma, S.J. (author), Goudriaan, M. (author), van Maris, A.J.A. (author), Pronk, J.T. (author)

Simultaneous fermentation of glucose and xylose can contribute to improved productivity and robustness of yeast-based processes for bioethanol production from lignocellulosic hydrolysates. This study explores a novel laboratory evolution strategy for identifying mutations that contribute to simultaneous utilisation of these sugars in batch...

journal article 2018

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Fermentation of glucose-xylose-arabinose mixtures by a synthetic consortium of single-sugar-fermenting Saccharomyces cerevisiae strains

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