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β-Carotene production by Saccharomyces cerevisiae with regard to plasmid stability and culture media

  • Applied Microbial and Cell Physiology
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Abstract

A recombinant Saccharomyces cerevisiae strain was used for the production of β-carotene. The episomal plasmid YEplac195YB/I/E was extended by a gene coding for the mevalonate kinase (mvaK1) from Staphylococcus aureus. The adh1 promoter was chosen for constitutive expression of mvaK1. The recombinant strain S. cerevisiae G175 (YEplac-CaroSA) synthesised β-carotene by expressing the carotenogenic genes of Xanthophyllomyces dendrorhous together with the mvaK1 gene. Cells of this strain were investigated for their carotenoid contents in YNB and YPD media. A corresponding mvaK1 transcript in the recombinant yeast host was verified. Growth experiments of a specific erg12 deletion mutant showed that the mevalonate kinase (MvaK1) was able to complement the function of the deleted native mevalonate kinase (Erg12) from S. cerevisiae in the MVA pathway under control of the constitutive adh1 promoter. Cells of S. cerevisiae G175 (YEplac-CaroSA) exhibited high plasmid stability under either selective or non-selective cultivation conditions. Time course experiments demonstrated high plasmid stability even over extended cultivation periods. Carotenoid production was therefore also stable in larger culture volumes. Due to the stability of the plasmid, cultivation of the cells in complex YPD medium was possible, and 14.3 mg β-carotene per litre and a cell density of 9 g cell dry matter (CDM) per litre were achieved. The highest amount of 3,897 μg β-carotene per gramme CDM at a cell density of 1 g CDM per litre was measured after cultivation of the cells in YNB medium with glucose as sole carbon source.

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Acknowledgements

We thank Rene Verwaal (DSM Biotechnology Centre, Delft, The Netherlands) for the generous provision of the yeast plasmids for β-carotene production. We thank Roland Klassen and Friedhelm Meinhardt (Institut für Molekulare Mikrobiologie und Biotechnologie, Münster, Germany) for providing vector pUG73 and Kai Müller (Fraunhofer Institut für Molekularbiologie und Angewandte Ökologie, Aachen, Germany) for providing vector PADNsβAS1. We also thank M. Gustavsson, E. Wiberg, P. Stolt and S. Stymne (Scandinavian Biotechnology Research, Alnarp, Sweden) for providing S. cerevisiae G175.

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  1. Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Münster, Corrensstraße 3, 48149, Münster, Germany

    Nicole Lange & Alexander Steinbüchel

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  1. Nicole Lange
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  2. Alexander Steinbüchel
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Correspondence to Alexander Steinbüchel.

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Lange, N., Steinbüchel, A. β-Carotene production by Saccharomyces cerevisiae with regard to plasmid stability and culture media. Appl Microbiol Biotechnol 91, 1611–1622 (2011). https://doi.org/10.1007/s00253-011-3315-2

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