Engineering precursor supply for the high-level production of ergothioneine in Saccharomyces cerevisiae.

van der Hoek, Steven A, Rusnák, Matej, Wang, Guokun, Stanchev, Lyubomir Dimitrov, de Fátima Alves, Luana, Jessop-Fabre, Mathew M, Paramasivan, Kalaivani, Jacobsen, Irene Hjorth, Sonnenschein, Nikolaus, Martínez, José L
et al (show 3 more authors) (2022) Engineering precursor supply for the high-level production of ergothioneine in Saccharomyces cerevisiae. Metabolic engineering, 70. pp. 129-142.

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Ergothioneine (ERG) is an unusual sulfur-containing amino acid. It is a potent antioxidant, which shows great potential for ameliorating neurodegenerative and cardiovascular diseases. L-ergothioneine is rare in nature, with mushrooms being the primary dietary source. The chemical synthesis process is complex and expensive. Alternatively, ERG can be produced by fermentation of recombinant microorganisms engineered for ERG overproduction. Here, we describe the engineering of S. cerevisiae for high-level ergothioneine production on minimal medium with glucose as the only carbon source. To this end, metabolic engineering targets in different layers of the amino acid metabolism were selected based on literature and tested. Out of 28 targets, nine were found to improve ERG production significantly by 10%-51%. These targets were then sequentially implemented to generate an ergothioneine-overproducing yeast strain capable of producing 106.2 ± 2.6 mg/L ERG in small-scale cultivations. Transporter engineering identified that the native Aqr1 transporter was capable of increasing the ERG production in a yeast strain with two copies of the ERG biosynthesis pathway, but not in the strain that was further engineered for improved precursor supply. Medium optimization indicated that additional supplementation of pantothenate improved the strain's productivity further and that no supplementation of amino acid precursors was necessary. Finally, the engineered strain produced 2.39 ± 0.08 g/L ERG in 160 h (productivity of 14.95 ± 0.49 mg/L/h) in a controlled fed-batch fermentation without supplementation of amino acids. This study paves the way for the low-cost fermentation-based production of ergothioneine.

Item Type: Article
Uncontrolled Keywords: Ergothioneine, Metabolic engineering, Medium optimization, Saccharomyces cerevisiae, Yeast, Nutraceutical
Divisions: Faculty of Health and Life Sciences
Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology
Depositing User: Symplectic Admin
Date Deposited: 18 May 2022 09:28
Last Modified: 18 Jan 2023 21:01
DOI: 10.1016/j.ymben.2022.01.012
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