title: Exploiting cell metabolism for biocatalytic whole-cell transamination by recombinant Saccharomyces cerevisiae.
abstract: The potential of Saccharomyces cerevisiae for biocatalytic whole-cell transamination was investigated using the kinetic resolution of racemic 1-phenylethylamine (1-PEA) to (R)-1-PEA as a model reaction. As native yeast do not possess any ω-transaminase activity for the reaction, a recombinant yeast biocatalyst was constructed by overexpressing the gene coding for vanillin aminotransferase from Capsicum chinense. The yeast-based biocatalyst could use glucose as the sole co-substrate for the supply of amine acceptor via cell metabolism. In addition, the biocatalyst was functional without addition of the co-factor pyridoxal-5'-phosphate (PLP), which can be explained by a high inherent cellular capacity to sustain PLP-dependent reactions in living cells. In contrast, external PLP supplementation was required when cell viability was low, as it was the case when using pyruvate as a co-substrate. Overall, the results indicate a potential for engineered S. cerevisiae as a biocatalyst for whole-cell transamination and with glucose as the only co-substrate for the supply of amine acceptor and PLP.
authors.affiliation: Division of Applied Microbiology, Department of Chemistry, Lund University, Getingevägen 60, 22100, Lund, Sweden.
authors.forename: Nora
authors.initials: N
authors.lastname: Weber
authors.forename: Marie
authors.initials: M
authors.lastname: Gorwa-Grauslund
authors.forename: Magnus
authors.initials: M
authors.lastname: Carlquist
mesh_terms: D002212:Capsicum / Q000201:enzymology / Q000235:genetics; D004798:Enzymes / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D010627:Phenethylamines / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D000637:Transaminases / Q000378:metabolism*
chemical_list: D004798:Enzymes; D010627:Phenethylamines; D011994:Recombinant Proteins; D000637:Transaminases; D005947:Glucose