Print Email Facebook Twitter Engineering Saccharomyces cerevisiae for fast vitamin-independent aerobic growth Title Engineering Saccharomyces cerevisiae for fast vitamin-independent aerobic growth Author Ehrmann, Anja K. (Technical University of Denmark) Wronska, A.K. (TU Delft BT/Industriele Microbiologie) Perli, T. (TU Delft BT/Industriele Microbiologie) de Hulster, A.F. (TU Delft BT/Industriele Microbiologie) Luttik, M.A.H. (TU Delft BT/Industriele Microbiologie) van den Broek, M.A. (TU Delft BT/Industriele Microbiologie) Carqueija Cardoso, D.C. (TU Delft BT/Industriele Microbiologie) Pronk, J.T. (TU Delft BT/Biotechnologie) Daran, J.G. (TU Delft BT/Industriele Microbiologie) Department BT/Biotechnologie Date 2024 Abstract 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 preparation and may decrease susceptibility to contamination by auxotrophic microbes. In this study, suboptimal growth rates of S. cerevisiae CEN.PK113-7D in the absence of pantothenic acid, para-aminobenzoic acid (pABA), pyridoxine, inositol and/or biotin were corrected by single or combined overexpression of ScFMS1, ScABZ1/ScABZ2, ScSNZ1/ScSNO1, ScINO1 and Cyberlindnera fabianii BIO1, respectively. Several strategies were explored to improve growth of S. cerevisiae CEN.PK113-7D in thiamine-free medium. Overexpression of ScTHI4 and/or ScTHI5 enabled thiamine-independent growth at 83% of the maximum specific growth rate of the reference strain in vitamin-supplemented medium. Combined overexpression of seven native S. cerevisiae genes and CfBIO1 enabled a maximum specific growth rate of 0.33 ± 0.01 h−1 in vitamin-free synthetic medium. This growth rate was only 17 % lower than that of a congenic reference strain in vitamin-supplemented medium. Physiological parameters of the engineered vitamin-independent strain in aerobic glucose-limited chemostat cultures (dilution rate 0.10 h−1) grown on vitamin-free synthetic medium were similar to those of similar cultures of the parental strain grown on vitamin-supplemented medium. Transcriptome analysis revealed only few differences in gene expression between these cultures, which primarily involved genes with roles in Class-B vitamin metabolism. These results pave the way for development of fast-growing vitamin-independent industrial strains of S. cerevisiae. To reference this document use: http://resolver.tudelft.nl/uuid:5611465d-9d0c-404e-afcd-587e9631e06f DOI https://doi.org/10.1016/j.ymben.2024.01.010 ISSN 1096-7176 Source Metabolic Engineering, 82, 201-215 Part of collection Institutional Repository Document type journal article Rights © 2024 Anja K. Ehrmann, A.K. Wronska, T. Perli, A.F. de Hulster, M.A.H. Luttik, M.A. van den Broek, D.C. Carqueija Cardoso, J.T. Pronk, J.G. Daran Files PDF 1-s2.0-S1096717624000168-main.pdf 5.78 MB Close viewer /islandora/object/uuid:5611465d-9d0c-404e-afcd-587e9631e06f/datastream/OBJ/view