title: Manipulation of the Precursor Supply in Yeast Significantly Enhances the Accumulation of Prenylated β-Carbolines.
abstract: The tryptophan derivative 1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid (MTCA) is present in many plants and foods including fermentation products of the baker's yeast Saccharomyces cerevisiae. MTCA is formed from tryptophan and acetaldehyde via a Pictet-Spengler reaction. In this study, up to 9 mg/L of MTCA were detected as a mixture of (1S,3S) and (1R,3S) isomers in a ratio of 2.2:1 in Saccharomyces cerevisiae cultures. To the best of our knowledge, this is the first report on the presence of MTCA in laboratory baker's yeast cultures. Expression of three fungal tryptophan prenyltransferase genes, fgaPT2, 5-dmats, and 7-dmats in S. cerevisiae resulted in the formation of MTCA derivatives with prenyl moieties at different positions of the indole ring. Expression of these genes in dimethylallyl diphosphate and tryptophan overproducing strains led to generation of up to 400 mg/L of prenylated MTCAs as mixtures of (1S,3S) and (1R,3S) diastereomers in ratios similar to that of unprenylated MTCA. The structures of the described substances including their stereochemistry were unequivocally elucidated by mass spectrometry as well as one- and two-dimensional NMR spectroscopy. The results of this study provide a convenient system for the production of high amounts of designed prenylated MTCAs in S. cerevisiae. Furthermore, our work can be considered as an excellent example for the construction of more complex molecules by introducing just one key gene.
authors.affiliation: Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg , Robert-Koch-Straße 4, 35037 Marburg, Germany.|Zentrum für Synthetische Mikrobiologie, Philipps-Universität Marburg , Hans-Meerwein-Straße, 35032 Marburg, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Backhaus
authors.affiliation: Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg , Robert-Koch-Straße 4, 35037 Marburg, Germany.
authors.forename: Lena
authors.initials: L
authors.lastname: Ludwig-Radtke
authors.affiliation: Fachbereich Chemie, Philipps-Universität Marburg , Hans-Meerwein-Straße, 35032 Marburg, Germany.
authors.forename: Xiulan
authors.initials: X
authors.lastname: Xie
authors.affiliation: Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg , Robert-Koch-Straße 4, 35037 Marburg, Germany.|Zentrum für Synthetische Mikrobiologie, Philipps-Universität Marburg , Hans-Meerwein-Straße, 35032 Marburg, Germany.
authors.forename: Shu-Ming
authors.identifier: 0000-0003-4583-2655
authors.initials: SM
authors.lastname: Li
mesh_terms: D002243:Carbolines / Q000032:analysis / Q000378:metabolism*; D018929:Cell Culture Techniques; D004122:Dimethylallyltranstransferase / Q000378:metabolism*; D060847:Metabolic Engineering; D054876:Prenylation; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D014364:Tryptophan / Q000378:metabolism
chemical_list: D002243:Carbolines; C038114:1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid; D014364:Tryptophan; D004122:Dimethylallyltranstransferase
keywords: 1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid; fungal tryptophan prenyltransferase; regiospecific prenylation; β-carboline