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Study on the construction and aroma-producing characteristics of the recombinant Saccharomyces cerevisiae strain W303-EAT

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Abstract

In this study, a recombinant Saccharomyces cerevisiae strain W303-EAT overexpressing the alcohol acetyltransferase (AAT) gene EAT was constructed by the homologous recombination principle. The results showed that, compared the recombinant strain W303-EAT to its parent W303-1A, there were no significant differences in the growth performance, CO2 loss, alcohol content, and reducing sugar content. Bioinformatics analysis results demonstrated that AAT protein is a hydrophobic, secreted, and non-transmembrane protein. Then, the differences in fermentation performance and aroma-producing characteristics between the recombinant strain W303-EAT and its parent strain W303-1A were studied by low-alcohol fermentation wines. The results showed that the rice wine, blueberry wine, and rose wine of the recombinant strain W303-EAT applied to the fermentation produced higher ethyl acetate than its parent strain W303-1A. Meanwhile, we found that the rice wine, blueberry wine, and rose wine fermented by the recombinant strain W303-EAT had lower contents of isobutanol, isoamylol, and phenethanol than its parent strain W303-1A, indicating that the recombinant strain W303-EAT had an inevitable effect of reducing higher alcohol contents. This study provided a better understanding of aroma-producing characteristics in recombinant Saccharomyces cerevisiae strain W303-EAT overexpressing the EAT gene.

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References

  1. Mitropoulou G, Nikolaou A, Santarmaki V, Sgouros G, Kourkoutas Y (2020) Citrus medica and Cinnamomum zeylanicum essential oils as potential biopreservatives against spoilage in low alcohol wine products. Foods 9(5):577

    Article  CAS  Google Scholar 

  2. Gobbi M, Comitini F, Domizio P, Romani C, Lencioni L, Mannazzu I (2010) Non-Saccharomyces yeasts in controlled mixed culture fermentation in winemaking: the role of metabolic interactions. J Biotechnol 150:299–300

    Article  Google Scholar 

  3. Vilanova M, Martínez C (2007) First study of determination of aromatic compounds of red wine from Vitis vinifera cv. Castañal grown in Galicia (NW Spain). Eur Food Res Technol 224:431–436

    Article  CAS  Google Scholar 

  4. Ivanova V, Stefova M, Vojnoski B, Stafilov T, Bíró I, Bufa A, Felinger A, Kilár F (2013) Volatile composition of Macedonian and Hungarian wines assessed by GC/MS. Food Bioprocess Tech 6:1609–1617

    Article  CAS  Google Scholar 

  5. Bai MY, Wu ZF, Li RY, Xiong GT, Weng PF (2019) The growth law of wine yeasts in a mixed culture and difference analysis of volatile flavor compounds. J Chin Inst Food Sci Technol 19:214–221

    Google Scholar 

  6. Liu CZ, Guo B, Bu AQ, Yu YQ, Zhou SS (2019) Flavor characteristics of fruit wine yeast and Wickerhamomyces anomalus yeast mixed fermentation. J Anhui Agric Sci 47(4):162-164+169

    Google Scholar 

  7. Zhang M, Shuai XX, Ma FY, Du LQ, Tu XH (2018) Optimization of fermentation condition for Manggo fruit wine and analysis of volatile flavor components. Guangdong Chem 21(45):23–26

    Google Scholar 

  8. Lu H, Du H, Xu Y (2012) Study on the change rules of earthy-musty odorants in solid fermentation and distillation process of Chinese liquor. Liquor-Mak Sci Technol 8:29–32

    Google Scholar 

  9. Kruis AJ, Gallone B, Jonker T, Mars AE, van Rijswijck IMH, Wolkers-Rooijackers JCM, Smid EJ, Steensels J, Verstrepen KJ, Kengen SWM, van der Oost J, Weusthuis RA (2018) Contribution of Eat1 and other alcohol acyltransferases to ester production in Saccharomyces cerevisiae. Front Microbiol 9:3202

    Article  Google Scholar 

  10. Park YC, Shaffer CEH, Bennett GN (2009) Microbial formation of esters. Appl Microbiol Biot 85:13–25

    Article  CAS  Google Scholar 

  11. Kruis AJ, Levisson M, Mars AE, van der Ploeg M, Daza FG, Ellena V, Kengen SWM, van der Oosta J, Weusthuis RA (2017) Ethyl acetate production by the elusive alcohol acetyltransferase from yeast. Metab Eng 41:92–101

    Article  CAS  Google Scholar 

  12. Gietz RD, Woods RA (2002) Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method. Methods Enzymol 350:87–96

    Article  CAS  Google Scholar 

  13. Liu YL, Zhang CY, Zhang Y, Wang DS, Xiao DG (2012) Fermentation performances of Saccharomyces cerevisiae with ATF1 overexpression and BAT2 knock-out. China Brew 31:151–153

    CAS  Google Scholar 

  14. Du YJ, Chen YF, Li J, He YH, Gong R, Guo XW, Xiao DG (2018) Effect of FAS2 gene overexpression on the production of favor-active. Mod Food Sci Tech 34(7):81–88

    CAS  Google Scholar 

  15. Wang FR (2005) Analysis and detection of wine. Chemical Industry Publisher, Beijing

    Google Scholar 

  16. Zhao K, Xu PJ, Gu GY (2008) Study on determination of reducing sugar content using 3,5-dinitrosalicylic acid method. Food Sci 29:534–536

    CAS  Google Scholar 

  17. Miao SQ, Yao WX (2015) Study on optimum fermentation conditions for rice wine. Liquor Making 42:67–69

    CAS  Google Scholar 

  18. Liu CT (2020) Study on fermentation process of blueberry wine. Guizhou University

    Google Scholar 

  19. Zheng SY, Wang HY, Yu WP, Hou DS, Kong QL, You Y (2017) Fermentation process of rose wine. Sci Technol Food Ind 38:114–118

    Google Scholar 

  20. Yoshioka K, Hashimoto N (1981) Ester formation by alcohol acetyltransferase from brewers’ yeast. Agric Bioi Chem 45:2183–2190

    CAS  Google Scholar 

  21. Plata C, Millán C, Mauricio JC, Ortega JM (2020) Formation of ethyl acetate and isoamyl acetate by various species of wine yeasts. Food Microbiol 20:217–224

    Article  Google Scholar 

  22. Fu ZL, Sun BG, Li XT, Fan GS, Teng C, Allaa A, Jia YM (2018) Isolation and characterization of a high ethyl acetate-producing yeast from Laobaigan Daqu and its fermentation conditions for producing high-quality Baijiu. Biotechnol Biotech Eq 32:1218–1227

    Article  CAS  Google Scholar 

  23. Kruis AJ, Mars AE, Kengen SWM, Borst JW, van der Oost J, Weusthuis RA (2018) Alcohol acetyltransferase Eat1 is located in yeast mitochondria. Appl Environ Microb 84(19):e01640-18

    Article  Google Scholar 

  24. Gallone B, Steensels J, Prahl T, Soriaga L, Saels V, Herrera-Malaver B, Merlevede A, Roncoroni M, Voordeckers K, Miraglia L, Teiling C, Steffy B, Taylor M, Schwartz A, Richardson T, White C, Baele G, Maere S, Verstrepen KJ (2016) Domestication and divergence of Saccharomyces cerevisiae beer yeasts. Cell 166:1397.e16-1410.e16

    Article  Google Scholar 

  25. Saerens SMG, Delvaux F, Verstrepen KJ, Van Dijck P, Thevelein JM, Delvaux FR (2008) Parameters affecting ethyl ester production by Saccharomyces cerevisiae during fermentation. Appl Environ Microb 74:454–461

    Article  CAS  Google Scholar 

  26. Kruis AJ, Bohnenkamp AC, Nap B, Nielsen J, Mars AE, Wijffels RH, van der Oost J, Kengen SWM, Weusthuis RA (2020) From Eat to trEat: engineering the mitochondrial Eat1 enzyme for enhanced ethyl acetate production in Escherichia coli. Biotechnol Biofuels 13:76

    Article  CAS  Google Scholar 

  27. Löbs AK, Engel R, Schwartz C, Flores A, Wheeldon I (2017) CRISPR–Cas9-enabled genetic disruptions for understanding ethanol and ethyl acetate biosynthesis in Kluyveromyces marxianus. Biotechnol Biofuels 10:164

    Article  Google Scholar 

  28. Holt S, Trindade de Carvalho B, Foulquié-Moreno MR, Thevelein JM (2018) Polygenic analysis in absence of major effector ATF1 unveils novel components in yeast flavor ester biosynthesis. MBio 9(4):e01279-18

    Article  Google Scholar 

  29. Quain D, Duffield M (1985) A metabolic function for higher alcohol production by yeast. J Inst Brew 91:123–125

    Google Scholar 

  30. Meilgaard M (1975) Flavor chemistry of beer: Part I: flavor interaction between principal volatiles. Tech Q Mast Brew Assoc Am 12:107–117

    CAS  Google Scholar 

  31. Procopio S, Qian F, Becker T (2011) Function and regulation of yeast genes involved in higher alcohol and ester metabolism during beverage fermentation. Eur Food Res Technol 233(5):721–729

    Article  CAS  Google Scholar 

  32. Verstrepen KJ, Van Laere SDM, Vanderhaegen BMP, Derdelinckx G, Dufour JP, Pretorius IS, Winderickx J, Thevelein JM, Delvaux FR (2003) Expression levels of the yeast alcohol acetyltransferase genes ATF1, Lg-ATF1, and ATF2 control the formation of a broad range of volatile esters. Appl Environ Microb 69(9):5228–5237

    Article  CAS  Google Scholar 

Download references

Funding

This research was supported by Science and Technology Platform and Talent Team Project of Guizhou Province, Grant number [2018]5251; Talent Base of Fermentation Engineering and Liquor Making in Guizhou Province, Grant number [2018]3; Zunyi City Innovative Talent Team Program Project, Grant number [2020]9; Excellent Young Scientific and Technological Talent Program, Grant number [2019]5645.

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Authors and Affiliations

  1. School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China

    Xiaodan Wang, Ke Shen, Pei Li, Hongxiang Zhou, Xiaoye Luo, Shuyi Qiu & Shidong Ban

  2. Guizhou Provincial Key Laboratory of Fermentation Engineering and Biological Pharmacy, Guizhou University, Guiyang, 550025, China

    Xiaodan Wang, Ke Shen, Pei Li, Hongxiang Zhou, Xiaoye Luo, Shuyi Qiu & Shidong Ban

  3. Guizhou Zhenjiu Brewing Co., Ltd., Zunyi, 563000, China

    Ke Shen & Shaopei Tang

  4. Qiandongnan Engineering and Technology Research Center for Comprehensive Utilization of National Medicine, Kaili University, Kaili, 556011, China

    Pei Li

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  1. Xiaodan Wang
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  2. Ke Shen
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  3. Shaopei Tang
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  4. Pei Li
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  6. Xiaoye Luo
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  7. Shuyi Qiu
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  8. Shidong Ban
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Contributions

XW and PL formal analysis, data curation, writing—original draft preparation. WX, HZ, XL, SQ, and SB conceptualization, writing—original draft preparation, funding acquisition. XW, KS, SQ, and SB investigation, methodology. XW, SQ, and SB conceptualization, funding acquisition. KS and ST formal analysis, data curation. HZ and XL resources. SB writing—reviewing and editing. XW and SQ supervision.

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Correspondence to Shidong Ban.

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Wang, X., Shen, K., Tang, S. et al. Study on the construction and aroma-producing characteristics of the recombinant Saccharomyces cerevisiae strain W303-EAT. Eur Food Res Technol 248, 447–456 (2022). https://doi.org/10.1007/s00217-021-03890-z

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  • DOI: https://doi.org/10.1007/s00217-021-03890-z

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