Title
Coupling engineering of Saccharomyces cerevisiae with medium optimization for the production of ergothioneine
Conference Dates
July 14-18, 2019
Abstract
Ergothioneine (ERG) is a naturally occurring, exogenous antioxidant that is nonetheless abundant in the human body. It has been shown both to reduce oxidative damage and to be involved in several diseases in vivo1,2. Therefore, ergothioneine is poised to take a place in the dietary supplement industry. Here we describe the engineering of the yeast Saccharomyces cerevisiae and subsequent medium optimization to produce ergothioneine by fermentation. After integrating combinations of biosynthetic pathways from different organisms, we screened yeast strains for their production of ERG. Next, the highest producing strain was engineered with ergothioneine transporters, and its amino acid metabolism was altered by knock-out of Tor1 or Yih1. The bottleneck for ergothioneine production was determined by integration of a second copy of the pathway enzymes. We also optimized the media composition for production of ergothioneine using yeast S. cerevisiae. Following these manipulations, we obtained a titer of 630 mg/l in fed-batch cultivation in bioreactors. This work shows that with further engineering of the strain, current chemical synthesis of ergothioneine could be replaced with a sustainable alternative. 1. Cheah, I. K. & Halliwell, B. Ergothioneine; antioxidant potential, physiological function and role in disease. Biochim. Biophys. Acta - Mol. Basis Dis. 1822, 784–793 (2012). 2. Halliwell, B., Cheah, I. K. & Tang, R. M. Y. Ergothioneine - a diet-derived antioxidant with therapeutic potential. FEBS Lett. (2018). doi:10.1002/1873-3468.13123
Recommended Citation
Steven Axel van der Hoek, Bala Krishna Prabhala, Karolina Zugaj, Mathias Bernfried Biron, Douglas Bruce Kell, Irina Borodina, and Behrooz Darbani, "Coupling engineering of Saccharomyces cerevisiae with medium optimization for the production of ergothioneine" in "Biochemical and Molecular Engineering XXI", Christina Chan, Michigan State University, USA Mattheos Koffas, RPI, USA Steffen Schaffer, Evonik Industries, Germany Rashmi Kshirsagar, Biogen, USA Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/biochem_xxi/7