Print Email Facebook Twitter In vivo recombination of Saccharomyces eubayanus maltose-transporter genes yields a chimeric transporter that enables maltotriose fermentation Title In vivo recombination of Saccharomyces eubayanus maltose-transporter genes yields a chimeric transporter that enables maltotriose fermentation Author Brouwers, N. (TU Delft BT/Industriele Microbiologie; TU Delft OLD BT/Cell Systems Engineering) Gorter de Vries, A.R. (TU Delft BT/Industriele Microbiologie; TU Delft OLD BT/Cell Systems Engineering) van den Broek, M.A. (TU Delft BT/Industriele Microbiologie; TU Delft OLD BT/Cell Systems Engineering) Weening, S.M. (TU Delft BT/Industriele Microbiologie; TU Delft OLD BT/Cell Systems Engineering) Elink Schuurman, Tom D. (Heineken Supply Chain) Kuijpers, Niels G.A. (Heineken Supply Chain) Pronk, J.T. (TU Delft BT/Industriele Microbiologie; TU Delft OLD BT/Cell Systems Engineering) Daran, J.G. (TU Delft BT/Industriele Microbiologie; TU Delft OLD BT/Cell Systems Engineering) Date 2019 Abstract 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 the subtelomeric MAL gene family, and requires transporters for maltotriose uptake. While S. eubayanus strain CBS 12357T harbors four SeMALT genes which enable uptake of the α-di-glucoside maltose, it lacks maltotriose transporter genes. In S. cerevisiae, sequence identity indicates that maltotriose and maltose transporters likely evolved from a shared ancestral gene. To study the evolvability of maltotriose utilization in S. eubayanus CBS 12357T, maltotriose-assimilating mutants obtained after UV mutagenesis were subjected to laboratory evolution in carbon-limited chemostat cultures on maltotriose-enriched wort. An evolved strain showed improved maltose and maltotriose fermentation in 7 L fermenter experiments on industrial wort. Whole-genome sequencing revealed a novel mosaic SeMALT413 gene, resulting from repeated gene introgressions by non-reciprocal translocation of at least three SeMALT genes. The predicted tertiary structure of SeMalT413 was comparable to the original SeMalT transporters, but overexpression of SeMALT413 sufficed to enable growth on maltotriose, indicating gene neofunctionalization had occurred. The mosaic structure of SeMALT413 resembles the structure of S. pastorianus maltotriose-transporter gene SpMTY1, which has high sequences identity to alternatingly S. cerevisiae MALx1, S. paradoxus MALx1 and S. eubayanus SeMALT3. Evolution of the maltotriose transporter landscape in hybrid S. pastorianus lager-brewing strains is therefore likely to have involved mechanisms similar to those observed in the present study. To reference this document use: http://resolver.tudelft.nl/uuid:205dff5d-8e75-4670-bc83-5dbc6a83d910 DOI https://doi.org/10.1371/journal.pgen.1007853 ISSN 1553-7390 Source PLoS Genetics (Print), 15 (4), e1007853 Part of collection Institutional Repository Document type journal article Rights © 2019 N. Brouwers, A.R. Gorter de Vries, M.A. van den Broek, S.M. Weening, Tom D. Elink Schuurman, Niels G.A. Kuijpers, J.T. Pronk, J.G. Daran Files PDF journal.pgen.1007853.pdf 2.26 MB Close viewer /islandora/object/uuid:205dff5d-8e75-4670-bc83-5dbc6a83d910/datastream/OBJ/view