title: Enhanced incorporation yield of cysteine for glutathione overproduction by fed-batch fermentation of Saccharomyces cerevisiae.
abstract: In the following work a high cell density fed-batch process with Saccharomyces cerevisiae coupled with a high efficient incorporation of cysteine for glutathione (GSH) overproduction was developed. Therefore, a feeding strategy based on the respiratory quotient (RQ) was applied to ensure high biomass (96.1g/l). Furthermore, the optimal cysteine concentration and time of cysteine addition were investigated. Low concentrations of cysteine at late fermentation phases resulted in relatively high incorporation yields of about 0.40mol/mol and maintained the physiology of cultivated yeast. By changing the cysteine feeding from standard single shot to continuous addition, an often observed cell specific toxicity, triggered by high cysteine concentrations, could be prevented and the cysteine incorporation yield (0.54±0.01mol/mol) and GSH content (1650.7±42.8mg/l; 1.76±0.08%) were maximized, respectively. The developed process was transferred from laboratory into pilot plant scale. Further, the reduced cell specific toxicity enabled the development of a repeated fed-batch procedure with a suitable performance concerning cysteine incorporation yield (0.40±0.1mol/mol), biomass (84.2±1.2g/l) and GSH content (1304.7±61.4mg/l).
authors.affiliation: Berlin University of Technology, Institute of Biotechnology, Berlin, Germany.
authors.forename: Eric
authors.initials: E
authors.lastname: Lorenz
authors.affiliation: Research and Teaching Institute for Brewing in Berlin, Department Bioprocess Engineering and Applied Microbiology, Berlin, Germany.
authors.forename: Maximilian
authors.initials: M
authors.lastname: Schmacht
authors.affiliation: Berlin University of Technology, Institute of Biotechnology, Berlin, Germany; Research and Teaching Institute for Brewing in Berlin, Department Bioprocess Engineering and Applied Microbiology, Berlin, Germany.
authors.forename: Ulf
authors.initials: U
authors.lastname: Stahl
authors.affiliation: Research and Teaching Institute for Brewing in Berlin, Department Bioprocess Engineering and Applied Microbiology, Berlin, Germany. Electronic address: m.senz@vlb-berlin.org.
authors.forename: Martin
authors.initials: M
authors.lastname: Senz
mesh_terms: D061249:Batch Cell Culture Techniques / Q000379:methods*; D018533:Biomass; D003545:Cysteine / Q000378:metabolism*; D005285:Fermentation; D005978:Glutathione / Q000378:metabolism*; D010865:Pilot Projects; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013997:Time Factors
chemical_list: D005978:Glutathione; D003545:Cysteine
keywords: Cysteine incorporation yield; Fed-batch strategy; Glutathione enriched yeast; Glutathione overproduction; Repeated fed-batch; Scale-up