Abstract
Yeasts can convert amino acids to flavor alcohols following the Ehrlich pathway, a reaction sequence comprising transamination, decarboxylation, and reduction. The alcohols can be further derivatized to the acetate esters by alcohol acetyl transferase. Using l-methionine as sole nitrogen source and at high concentration, 3-(methylthio)-1-propanol (methionol) and 3-(methylthio)-propylacetate (3-MTPA) were produced with Saccharomyces cerevisiae. Methionol and 3-MTPA acted growth inhibiting at concentrations of >5 and >2 g L−1, respectively. With the wild type strain S. cerevisiae CEN.PK113-7D, 3.5 g L−1 methionol and trace amounts of 3-MTPA were achieved in a bioreactor. Overexpression of the alcohol acetyl transferase gene ATF1 under the control of a TDH3 (glyceraldehyde-3-phosphate dehydrogenase) promoter together with an optimization of the glucose feeding regime led to product concentrations of 2.2 g L−1 3-MTPA plus 2.5 g L−1 methionol. These are the highest concentrations reported up to now for the biocatalytic synthesis of these flavor compounds which are applied in the production of savory aroma compositions such as meat, potato, and cheese flavorings.
Similar content being viewed by others
References
Aoki T, Uchida K (1991) Enhanced formation of 3-(methylthio-)-1-propanol in a salt-tolerant yeast, Zygosaccharomyces rouxii, due to deficiency of S-adenosylmethionine synthase. Agric Biol Chem 55:2113–2116
Arctander S (1969) Perfume and flavor chemicals. Aroma chemicals. Published by the author, Montclair, USA
Arfi K, Landaud S, Bonnarme P (2006) Evidence for distinct L-methionine catabolic pathways in the yeast Geotrichum candidum and the bacterium Brevibacterium linens. Appl Environ Microbiol 72:2155–62
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1989) Current protocols in molecular biology. Wiley, New York
Ballance PE (1961) Production of volatile compounds related to the flavour of foods from the Strecker degradation of DL-methionine. J Sci Food Agric 12:532–536
Berry DR, Brown C (1987) Physiology of yeast growth. In: Berry DR, Russel I, Steward GG (eds) Yeast biotechnology. Allen and Unwin, London, pp 159–199
Bonnarme P, Psoni L, Spinnler HE (2000) Diversity of L-methionine catabolism pathways in cheese-ripening bacteria. Appl Environ Microbiol 66:5514–7
Burdock GA (2004) Fenaroli’s handbook of flavor ingredients. CRC, Boca Raton, p 1320
Dickinson JR, Lanterman MM, Danner DJ, Pearson BM, Sanz P, Harrison SJ, Hewlins MJ (1997) A 13C nuclear magnetic resonance investigation of the metabolism of leucine to isoamyl alcohol in Saccharomyces cerevisiae. J Biol Chem 272:26871–8
Dickinson JR, Harrison SJ, Hewlins MJ (1998) An investigation of the metabolism of valine to isobutyl alcohol in Saccharomyces cerevisiae. J Biol Chem 273:25751–6
Dickinson JR, Harrison SJ, Dickinson JA, Hewlins MJ (2000) An investigation of the metabolism of isoleucine to active Amyl alcohol in Saccharomyces cerevisiae. J Biol Chem 275:10937–42
Dickinson JR, Salgado LE, Hewlins MJ (2003) The catabolism of amino acids to long chain and complex alcohols in Saccharomyces cerevisiae. J Biol Chem 278:8028–34
van Dijken JP, Bauer J, Brambilla L, Duboc P, Francois JM, Gancedo C, Giuseppin ML, Heijnen JJ, Hoare M, Lange HC, Madden EA, Niederberger P, Nielsen J, Parrou JL, Petit T, Porro D, Reuss M, van Riel N, Rizzi M, Steensma HY, Verrips CT, Vindelov J, Pronk JT (2000) An interlaboratory comparison of physiological and genetic properties of four Saccharomyces cerevisiae strains. Enzyme Microb Technol 26:706–714
Entian K-D, Kötter P (2007) 25 Yeast genetic strain and plasmid collections. Meth Microbiol 36:629–666
Etschmann MMW, Sell D, Schrader J (2003) Screening of yeasts for the production of the aroma compound 2-phenylethanol in a molasses-based medium. Biotech Lett 25:531–536
Etschmann MMW, Sell D, Schrader J (2004) Medium optimization for the production of the aroma compound 2-phenylethanol using a genetic algorithm. J Mol Catal B 29:187–193
Etschmann MM, Sell D, Schrader J (2005) Production of 2-phenylethanol and 2-phenylethylacetate from L-phenylalanine by coupling whole-cell biocatalysis with organophilic pervaporation. Biotechnol Bioeng 92:624–634
Etschmann MM, Schrader J (2006) An aqueous-organic two-phase bioprocess for efficient production of the natural aroma chemicals 2-phenylethanol and 2-phenylethylacetate with yeast. Appl Microbiol Biotechnol 71:440–443
Gijs L, Perpète P, Timmermans A, Collin S (2000) 3-Methylthiopropionaldehyde as precursor of dimethyl trisulfide in aged beers. J Agric Food Chem 48:6196–6199
Güldener US, Heck T, Fielder J, Beinhauer, Hegemann JH (1996) A new efficient gene disruption cassette for repeated use in budding yeast. Nucleic Acids Res 24:2519–24
Hayashibe M, Katoda S, Owada H, Yoshida H, Katayosa A, Terashima T (1970) Methionine metabolism in yeast. J Ferment Technol 48:22–28
Hazelwood LA, Daran JM, van Maris AJ, Pronk JT, Dickinson JR (2008) The Ehrlich pathway for fusel alcohol production: a century of research on Saccharomyces cerevisiae metabolism. Appl Environ Microbiol 74:2259–66
Hirosawa I, Aritomi K, Hoshida H, Kashiwagi S, Nishizawa Y, Akada R (2004) Construction of a self-cloning sake yeast that overexpresses alcohol acetyltransferase gene by a two-step gene replacement protocol. Appl Microbiol Biotechnol 65:68–73
Hoffman CS, Winston F (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57:267–272
Kagkli DM, Tache R, Cogan TM, Hill C, Casaregola S, Bonnarme P (2006) Kluyveromyces lactis and Saccharomyces cerevisiae, two potent deacidifying and volatile-sulphur-aroma-producing microorganisms of the cheese ecosystem. Appl Microbiol Biotechnol 73:434–442
Kumar D, Gomes J (2005) Methionine production by fermentation. Biotechnol Adv 23:41–61
Landaud S, Helinck S, Bonnarme P (2008) Formation of volatile sulfur compounds and metabolism of methionine and other sulfur compounds in fermented food. Appl Microbiol Biotechnol 77:1191–205
Lilly M, Lambrechts MG, Pretorius IS (2000) Effect of increased yeast alcohol acetyltransferase activity on flavor profiles of wine and distillates. Appl Environ Microbiol 66:744–753
Liu SQ, Crow VL (2007) Dairy product and process. WO 2007/136280A1
Lusk JL, Rozan A (2005) Consumer acceptance of biotechnology and the role of second generation technologies in the USA and Europe. Trends Biotechnol 23:386–387
Moreira N, Mendes F, Hogg T, Vasconcelos I (2005) Alcohols, esters and heavy sulphur compounds production by pure and mixed cultures of apiculate wine yeasts. Int J Food Microbiol 103:285–294
Mueller DA (2007) Flavours: the legal framework. In: Berger RG (ed) Flavours and fragrances. Chemistry, bioprocessing and sustainability. Springer, Berlin, pp 15–24
Perpete P, Duthoit O, De Maeyer S, Imray L, Lawton AI, Stavropoulos KE, Gitonga VW, Hewlins MJ, Dickinson JR (2006) Methionine catabolism in Saccharomyces cerevisiae. FEMS Yeast Res 6:48–56
Schiestl RH, Gietz RD (1989) High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet 16:339–46
Schrader J, Etschmann MM, Sell D, Hilmer JM, Rabenhorst J (2004) Applied biocatalysis for the synthesis of natural flavour compounds—current industrial processes and future prospects. Biotechnol Lett 26:463–72
Schreiber WL, Scharpf LG, Katz I (1997) Future needs of chemistry in flavors and fragrances. Perfumer & Flavorist 22:11–16
Schreier P, Drawert FD, Junker A, Barton H, Leupold G (1976) Über die Biosynthese von Aromastoffen durch Mikroorganismen. Z Lebensm Unters Forsch 162:279–284
Seward R, Willetts JM, Dinsdale MG, Lloyd D (1996) The effects of ethanol, hexan-1-ol, and 2-phenylethanol on cider yeast growth, viability, and energy status; synergistic inhibition. J Inst Brew 102:439–443
Stark D, Münch T, Sonnleitner B, Marison IW, Stockar von U (2002) Extractive bioconversion of 2-phenylethanol from L-phenylalanine by Saccharomyces cerevisiae. Biotechnol Prog 18:514–523
Wach A, Brachat A, Pohlmann R, Philippsen P (1994) New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae. Yeast 10:1793–808
Whitehead IM, Ohleyer E (1993) Microbial carboxylic acid production method. WO9308293
Acknowledgements
We thank Dr. Rinji Akada for the donation of the yeast strain RAK 1857, R.C. Treatt & Co. Ltd. for the donation of the 3-MTPA standard, and the German Federal Ministry for Food, Agriculture and Consumer Protection for funding the project (No. 22008803).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Etschmann, M.M.W., Kötter, P., Hauf, J. et al. Production of the aroma chemicals 3-(methylthio)-1-propanol and 3-(methylthio)-propylacetate with yeasts. Appl Microbiol Biotechnol 80, 579–587 (2008). https://doi.org/10.1007/s00253-008-1573-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-008-1573-4