European Food Research and Technology

, Volume 236, Issue 6, pp 1009–1014 | Cite as

Increased esters and decreased higher alcohols production by engineered brewer’s yeast strains

  • Cui-Ying Zhang
  • Yu-Lan Liu
  • Ya-Nan Qi
  • Jian-Wei Zhang
  • Long-Hai Dai
  • Xue Lin
  • Dong-Guang XiaoEmail author
Original Paper


Esters and higher alcohols produced by yeast during the fermentation of wort have the greatest impact on the smell and taste of beer. Alcohol acetyltransferase, which is mainly encoded by the ATF1 gene, is one of the most important enzymes for acetate ester synthesis. Cytosolic branched-chain amino acid aminotransferase, on the other hand, which is encoded by the BAT2 gene, plays an important role in the production of branched-chain alcohols. The objective of this study is to construct engineered brewer’s yeast strains that produce more acetate esters and less higher alcohols. Industrial brewer’s yeast strain S5 was used as the parental strain to construct ATF1 overexpression and BAT2 deletion mutants. The engineered strains S5-2 and S5-4, which feature partial BAT2 allelic genes replaced by the constructed ATF1 overexpression cassette, were obtained. The ester production of the engineered strains was observed to increase significantly compared with that of the parental cells. The concentrations of ethyl acetate produced by the engineered strains S5-2 and S5-4 increased to 78.88 and 117.40 mg L−1, respectively, or about 7.7-fold and 11.5-fold higher than that produced by parental S5 cells. The isoamyl acetate produced by S5-2 and S5-4 also increased to 5.14 and 9.25 mg L−1, respectively; by contrast, no isoamyl acetate was detected in the fermentation sample of the parental strain S5. Moreover, S5-2 and S5-4, respectively, produced about 65 and 51 % of higher alcohols produced by the parental strain. The increase in acetate ester content and decrease in higher alcohol concentration shown by the engineered brewer’s yeast strains at the end of fermentation process indicate that the new strains are useful in future developments in the wheat beer industry.


Saccharomyces cerevisiae Higher alcohols Acetate ester Branched-chain amino acids aminotransferase Alcohol acetyltransferase 



This work was supported by the National Natural Science Foundation of China (31000043) and Major Project of Research Program on Applied Fundamentals and Advanced Technologies of Tianjin (10JCZDJC16700).

Conflict of interest


Compliance with Ethics Requirements

This article does not contain any studies with human or animal subjects.


  1. 1.
    Styger G, Prior B, Bauer FF (2011) Wine flavor and aroma. J Ind Microbiol Biotechnol 38:1145–1159CrossRefGoogle Scholar
  2. 2.
    Swiegers JH, Pretorius IS (2005) Yeast modulation of wine flavour. Adv Appl Microbiol 57:131–175CrossRefGoogle Scholar
  3. 3.
    Swiegers JH, Bartowsky EJ, Henschke PA, Pretorius IS (2005) Yeast and bacterial modulation of wine aroma and flavour. Aust J Grape Wine Res 11:139–173CrossRefGoogle Scholar
  4. 4.
    Amerine AM, Berg HV, Kunkee RE, Ough CS, Singleton VL, Webb AD (1980) The technology of winemaking, 4th edn. AVI Technical Books Inc, WestportGoogle Scholar
  5. 5.
    Styger G, Jacobson D, Bauer FF (2011) Identifying genes that impact on aroma profiles produced by Saccharomyces cerevisiae and the production of higher alcohols. Appl Microbiol Biotechnol 91(3):713–730CrossRefGoogle Scholar
  6. 6.
    Saerens SM, Delvaux FR, Verstrepen KJ, Thevelein JM (2010) Production and biological function of volatile esters in Saccharomyces cerevisiae. Microb Biotechnol 3(2):165–177CrossRefGoogle Scholar
  7. 7.
    Meilgaard MC (2001) Effects on flavour of innovations in brewery equipment and processing: a review. J Inst Brew 107:271–286CrossRefGoogle Scholar
  8. 8.
    Meilgaard MC (1975) Flavour chemistry of beer Flavour and threshold of 239 aroma volatiles. MBAA Tech Q 12:151–168Google Scholar
  9. 9.
    Meilgaard MC (1975) Flavour chemistry of beer Flavour interaction between principal volatiles. MBAA Tech Q 12:107–117Google Scholar
  10. 10.
    Verstrepen KJ, Moonjai N, Derdelinckx G, Dufour JP, Winderickx J, Thevelein JM, Pretorius IS, Delvaux FR (2003) Genetic regulation of ester synthesis in brewer’s yeast: new facts, insights and implications for the brewer. In: Smart K (ed) Brewing yeast fermentation performance, vol 2, 2nd edn. Blackwell Science, Oxford, pp 234–248Google Scholar
  11. 11.
    Rankine BC (1967) Formation of higher alcohols by wine yeasts, and relationship to taste thresholds. J Sci Food Agric 18:583–589CrossRefGoogle Scholar
  12. 12.
    Saerens SM, Verbelen PJ, Vanbeneden N, Thevelein JM, Delvaux FR (2008) Monitoring the influence of high-gravity brewing and fermentation temperature on flavour formation by analysis of gene expression levels in brewing yeast. Appl Microbiol Biotechnol 80(6):1039–1051CrossRefGoogle Scholar
  13. 13.
    Sentheshanmuganathan S (1956) The formation of tyrosol (2-p-hydroxyethyl) from tyrosine by Saccharomyces cerevisiae. Biochem J 64:37–38Google Scholar
  14. 14.
    Sentheshanmuganathan S (1960) The mechanism of formation of higher alcohols from amino acids by Saccharomyces cerevisiae. Biochem J 74:568–576Google Scholar
  15. 15.
    Eden A, Benvenisty N (1998) Characterization of a branched-chain amino-acid aminotransferase from Schizosaccharomyces pombe. Yeast 14:189–194CrossRefGoogle Scholar
  16. 16.
    Eden A, Simchen G, Benvenisty N (1996) Two yeast homologs of ECA39, a target for c-Myc regulation, code for cytosolic and mitochondrial branched-chain amino acid aminotransferases. J Biol Chem 271(34):20242–20245CrossRefGoogle Scholar
  17. 17.
    Kispal G, Steiner H, Court DA, Rolinski B, Lill R (1996) Mitochondrial and cytosolic branched-chain amino acid transaminases from yeast, homologs of the myc oncogene-regulated Eca39 protein. J Biol Chem 271(40):24458–24464CrossRefGoogle Scholar
  18. 18.
    Lilly M, Bauer FF, Styger G, Lambrechts MG, Pretorius IS (2006) The effect of increased branched-chain amino acid transaminase activity in yeast on the production of higher alcohols and on the flavour profiles of wine and distillates. FEMS Yeast Res 6(5):726–743CrossRefGoogle Scholar
  19. 19.
    Eden A, Van Nedervelde L, Drukker M, Benvenisty N, Debourg A (2001) Involvement of branched-chain amino acid aminotransferases in the production of fusel alcohols during fermentation in yeast. Appl Microbiol Biotechnol 55(3):296–300CrossRefGoogle Scholar
  20. 20.
    Nordström K (1964) Formation of esters from alcohols by brewer’s yeast. J Inst Brew 70:328–336CrossRefGoogle Scholar
  21. 21.
    Nordström K (1962) Formation of ethyl acetate in fermentation with brewer’s yeast III: participation of coenzyme A. J Inst Brew 68:398–407CrossRefGoogle Scholar
  22. 22.
    Nordström K (1963) Formation of ethyl acetate in fermentation with brewer’s yeast IV: metabolism of acetyl coenzyme A. J Inst Brew 69:142–153CrossRefGoogle Scholar
  23. 23.
    Lilly M, Bauer FF, Lambrechts MG, Swiegers JH, Cozzolino D, Pretorius IS (2006) The effect of increased yeast alcohol acetyltransferase and esterase activity on the flavour profiles of wine and distillates. Yeast 23(9):641–659CrossRefGoogle Scholar
  24. 24.
    Fujii T, Nagasawa N, Iwamatsu A, Bogaki T, Tamai Y, Hamachi M (1994) Molecular cloning, sequence analysis and expression of the yeast alcohol acetyltransferase gene. Appl Environ Microbiol 60:2786–2792Google Scholar
  25. 25.
    Fujii T, Yoshimoto H, Nagasawa N, Bogaki T, Tamai Y, Hamachi M (1996) Nucleotide sequence of alcohol acetyltransferase genes from lager brewing yeast, Saccharomyces carlsbergensis. Yeast 12:593–598CrossRefGoogle Scholar
  26. 26.
    Yoshimoto H, Fujiwara D, Momma T, Tanaka K, Sone H, Nagasawa N, Tamai T (1999) Isolation and characterization of the ATF2 gene encoding alcohol acetyltransferase II in the bottom fermenting yeast Saccharomyces pastorianus. Yeast 15:409–417CrossRefGoogle Scholar
  27. 27.
    Yoshimoto H, Momma T, Fujiwara D, Sone H, Kaneko Y, Tamai T (1998) Characterization of the ATF1 and Lg-ATF1 genes encoding alcohol acetyltransferases in the bottom fermenting yeast Saccharomyces pastorianus. J Ferment Bioeng 86:15–20CrossRefGoogle Scholar
  28. 28.
    Lilly M, Lambrechts MG, Pretorius IS (2000) Effect of increased yeast alcohol acetyltransferase activity on flavour profiles of wine and distillates. Appl Environ Microbiol 66:744–753CrossRefGoogle Scholar
  29. 29.
    Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1994) Current protocols in molecular biology. Wiley, New YorkGoogle Scholar
  30. 30.
    Schiestl RH, Gietz RD (1989) High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet 16(5–6):339–346CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Cui-Ying Zhang
    • 1
  • Yu-Lan Liu
    • 1
  • Ya-Nan Qi
    • 1
  • Jian-Wei Zhang
    • 1
  • Long-Hai Dai
    • 1
  • Xue Lin
    • 1
  • Dong-Guang Xiao
    • 1
    Email author
  1. 1.Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, College of BiotechnologyTianjin University of Science and TechnologyTianjinPeople’s Republic of China

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