Screening of aroma-producing Saccharomyces cerevisiae strains and their effects on the flavor profiles of mechanized Huangjiu

Abstract

Mechanized Huangjiu (Chinese rice wine) is characterized by a stable flavor quality and short fermentation time, yet its flavor quality is inferior to that of traditional Huangjiu. Given the significant market demand for high-quality mechanized Huangjiu and the existing flavor quality gap, this study aimed to isolate and identify highly efficient aroma-producing yeasts from the fermentation mash of traditional Huangjiu for application in mechanized Huangjiu production. From 26 isolated strains, three Saccharomyces cerevisiae strains (OS7302, q1, and NL9) were selected based on their high aroma production capacity in simulated fermentation media. Gas chromatography-mass spectrometry (GC–MS), gas chromatography-olfactometry (GC–O), sensory evaluation, and physicochemical analysis were then combined to systematically verify their abilities to produce aroma compounds. In co-fermentation with mechanized starter cultures, S. cerevisiae OS7302 induced significant increases in the concentrations and aroma intensities of key esters, including ethyl hexanoate, ethyl caprate, and ethyl caprylate, thereby generating an ester aroma profile of mechanized Huangjiu comparable to that of traditional Huangjiu. Furthermore, Huangjiu fermented with S. cerevisiae OS7302 exhibited more intense fruit and ester aromas. In contrast, strains q1 and NL9 promoted the formation of higher alcohols. The use of aroma-producing S. cerevisiae as adjunct cultures for co-fermentation with mechanized starter cultures can improve the flavor quality of mechanized Huangjiu, alleviating the flavor gap between mechanized and traditional Huangjiu.

References

[1] Chen C, Tian T, Yu H, Yuan H, Wang B, et al. 2022. Characterisation of the key volatile compounds of commercial Gouda cheeses and their contribution to aromas according to Chinese consumers' preferences. Food Chemistry: X 15: 100416

[2] Tian S, Zeng W, Fang F, Zhou J, Du G. 2022. The microbiome of Chinese rice wine (Huangjiu). Current Research in Food Science 5: 325-335

[3] Yu H, Li Q, Guo W, Chen C, Ai L, et al. 2023. Dynamic analysis of volatile metabolites and microbial community and their correlations during the fermentation process of traditional Huangjiu (Chinese rice wine) produced around Winter Solstice . Food Chemistry: X 18: 100620

[4] Yang Y, Xia Y, Yan X, Li S, Ni L, et al. 2023. Insights into whereby raw wheat Qu contributes to the flavor quality of Huangjiu during brewing. LWT 178: 114619

[5] Liu S, Chen Q, Zou H, Yu Y, Zhou Z, et al. 2019. A metagenomic analysis of the relationship between microorganisms and flavor development in Shaoxing mechanized huangjiu fermentation mashes. International Journal of Food Microbiology 303: 9-18

[6] Xie G, Zheng H, Qiu Z, Lin Z, Peng Q, et al. 2021. Study on relationship between bacterial diversity and quality of Huangjiu (Chinese Rice Wine) fermentation. Food Science & Nutrition 9: 3885-3892

[7] Xie GF, Cai JH, Qian B, Wang L, Lu ZD, et al. 2020. Changes of flavor components during mechanized brewing process of dry Huangjiu fermented in tank. Food and Fermentation Industries 46: 157-164 (in Chinese)

[8] Peng Q, Meng K, Zheng H, Yu H, Zhang Y, et al. 2022. Metabolites comparison in post—fermentation stage of manual (mechanized) Chinese Huangjiu (yellow rice wine) based on GC—MS metabolomics. Food Chemistry: X 14: 100324

[9] Yang Y, Xia Y, Wang G, Yu J, Ai L. 2017. Effect of mixed yeast starter on volatile flavor compounds in Chinese rice wine during different brewing stages. LWT 78: 373-381

[10] Gao Y, Wang Y, Cui F, Yin S, Hu F. 2024. The formation, influencing factors, efficacy, and analytical techniques of flavor substances in Huangjiu. Journal of Food Composition and Analysis 135: 106585

[11] Huang ZR, Hong JL, Xu JX, Li L, Guo WL, et al. 2018. Exploring core functional microbiota responsible for the production of volatile flavour during the traditional brewing of Wuyi Hong Qu glutinous rice wine . Food Microbiology 76: 487-496

[12] Zhao Y, Liu S, Yang Q, Han X, Zhou Z, et al. 2022. Saccharomyces cerevisiae strains with low—yield higher alcohols and high—yield acetate esters improve the quality, drinking comfort and safety of huangjiu . Food Research International 161: 111763

[13] Yang Y, Xia Y, Lin X, Wang G, Zhang H, et al. 2018. Improvement of flavor profiles in Chinese rice wine by creating fermenting yeast with superior ethanol tolerance and fermentation activity. Food Research International 108: 83-92

[14] Ma D, Yuan L, Mao J, Liu T, Zhao Y, et al. 2025. Optimizing Huangjiu fermentation for enhanced aroma: insights into Saccharomyces cerevisiae jiangnan1# strain . Journal of Food Composition and Analysis 139: 107051

[15] Yang H, Qu T, Liu J, Zhang C, Chen M. 2025. Aroma—producing Saccharomyces cerevisiae improved the quality of Dangshan pear (Pyrus spp.) wine through the biosynthesis of key esters . Food Bioscience 69: 106799

[16] Svedlund N, Kiepert A, Rodríguez Pérez J, Magalhães F, Wietstock PC, et al. 2025. Enhancement of dry—hopped cider aroma through selection of apple cultivar, hop variety and yeast strain. Food Microbiology 129: 104755

[17] Guo W. 2022. Study on changes of community structure and aroma substances during fermentation of traditional Shaoxing Huangjiu under different ambient temperature conditions . Thesis. Shanghai Institute of Technology, Chinese. pp. 43-44 (in Chinese) doi: 10.27801/d.cnki.gshyy.2022.000179

[18] Huang X, Fan Y, Meng J, Sun S, Wang X, et al. 2021. Laboratory—scale fermentation and multidimensional screening of lactic acid bacteria from Daqu . Food Bioscience 40: 100853

[19] Yu H, Guo W, Xie T, Ai L, Tian H, et al. 2021. Aroma characteristics of traditional Huangjiu produced around Winter Solstice revealed by sensory evaluation, gas chromatography—mass spectrometry and gas chromatography—ion mobility spectrometry . Food Research International 145: 110421

[20] Yu H, Xie T, Xie J, Ai L, Tian H. 2019. Characterization of key aroma compounds in Chinese rice wine using gas chromatography—mass spectrometry and gas chromatography—olfactometry. Food Chemistry 293: 8-14

[21] Wang L, Fan S, Yan Y, Yang L, Chen S, et al. 2020. Characterization of potent odorants causing a pickle—like off—odor in Moutai—aroma type Baijiu by comparative aroma extract dilution analysis, quantitative measurements, aroma addition, and omission studies. Journal of Agricultural and Food Chemistry 68: 1666-1677

[22] Zhao Y, Liu S, Yang Q, Liu X, Xu Y, et al. 2023. Effects of simultaneous inoculation of non— Saccharomyces yeasts and Saccharomyces cerevisiae jiangnan1# on overall quality, flavor compounds, and sensory analysis of huangjiu . Food Bioscience 53: 102539

[23] 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. Applied Microbiology and Biotechnology 55: 296-300

[24] Zhou Z, Jiang Z, Zhang Z, Liu S, Ji Z, et al. 2024. Identification and interaction of the key odorants in Jimo Huangjiu by means of molecular sensory science. Journal of Food Composition and Analysis 135: 106695

[25] Molina AM, Swiegers JH, Varela C, Pretorius IS, Agosin E. 2007. Influence of wine fermentation temperature on the synthesis of yeast—derived volatile aroma compounds. Applied Microbiology and Biotechnology 77: 675-687

[26] Zhang B, Wang J, Jiang X, Huang M, Liu H, et al. 2024. Comparative study on key odorants of Jiujiang Fenggang Huangjiu and their succession regularities during aging using sensory—directed flavor analysis. Food Chemistry 430: 137052

[27] Fan W, Qian MC. 2005. Headspace solid phase microextraction and gas chromatography−olfactometry dilution analysis of young and aged Chinese "Yanghe Daqu" liquors. Journal of Agricultural and Food Chemistry 53: 7931-7938

[28] Chen GM, Huang ZR, Wu L, Wu Q, Guo WL, et al. 2021. Microbial diversity and flavor of Chinese rice wine (Huangjiu): an overview of current research and future prospects . Current Opinion in Food Science 42: 37-50

[29] Hu K, Jin GJ, Mei WC, Li T, Tao YS. 2018. Increase of medium—chain fatty acid ethyl ester content in mixed H. uvarum/ S. cerevisiae fermentation leads to wine fruity aroma enhancement . Food Chemistry 239: 495-501

[30] Pires EJ, Teixeira JA, Brányik T, Vicente AA. 2014. Yeast: the soul of beer's aroma—a review of flavour—active esters and higher alcohols produced by the brewing yeast. Applied Microbiology and Biotechnology 98: 1937-1949

[31] Zhao W, Ruan F, Qian M, Huang X, Li X, et al. 2023. Comparing the differences of physicochemical properties and volatiles in semi—dry Hakka rice wine and traditional sweet rice wine via HPLC, GC—MS and E—tongue analysis. Food Chemistry: X 20: 100898

[32] Fu W, Wang S, Xue W. 2024. Mechanism of carbohydrate and protein conversion during sourdough fermentation: an analysis based on representative Chinese sourdough microbiota. International Journal of Food Microbiology 410: 110487

[33] Wang P, Mao J, Meng X, Li X, Liu Y, et al. 2014. Changes in flavour characteristics and bacterial diversity during the traditional fermentation of Chinese rice wines from Shaoxing region. Food Control 44: 58-63

[34] Chen S, Xu Y, Qian MC. 2018. Comparison of the aromatic profile of traditional and modern types of Huang Jiu (Chinese rice wine) by aroma extract dilution analysis and chemical analysis . Flavour and Fragrance Journal 33: 263-271

[35] Liu SQ, Holland R, Crow VL. 2004. Esters and their biosynthesis in fermented dairy products: a review. International Dairy Journal 14: 923-945

[36] Bordet F, Romanet R, Bahut F, Ballester J, Eicher C, et al. 2023. Expanding the diversity of Chardonnay aroma through the metabolic interactions of Saccharomyces cerevisiae cocultures . Frontiers in Microbiology 13: 1032842

[37] Ye Y, Wang L, Zhan P, Tian H, Liu J. 2022. Characterization of the aroma compounds of Millet Huangjiu at different fermentation stages. Food Chemistry 366: 130691

[38] Grosch W. 1994. Determination of potent odourants in foods by aroma extract dilution analysis (AEDA) and calculation of odour activity values (OAVs). Flavour and Fragrance Journal 9: 147-158

[39] Van Gemert LJ. 2003. Compilations of odour threshold values in air, water and other media . Utrecht: Oliemans Punter & Partners BV. 352 pp.

[40] Luo T, Fan W, Xu Y. 2008. Characterization of volatile and semi—volatile compounds in Chinese rice wines by headspace solid phase microextraction followed by gas chromatography—mass spectrometry. Journal of the Institute of Brewing 114: 172-179

[41] Mo X, Fan W, Xu Y. 2009. Changes in volatile compounds of Chinese rice wine wheat Qu during fermentation and storage. Journal of the Institute of Brewing 115: 300-307

[42] Hu Q, Huang Z, Cheng S, Quan L, Wang L, et al. 2026. Advances in the study of bitter compounds in alcoholic beverages: a review and implications for the production of Chinese Huangjiu. Journal of Food Composition and Analysis 151: 108961

[43] Mao X, Yue SJ, Xu DQ, Fu RJ, Han JZ, et al. 2023. Research progress on flavor and quality of Chinese rice wine in the brewing process. ACS Omega 8: 32311-32330

[44] Carrau FM, Medina K, Farina L, Boido E, Henschke PA, et al. 2008. Production of fermentation aroma compounds by Saccharomyces cerevisiae wine yeasts: effects of yeast assimilable nitrogen on two model strains . FEMS Yeast Research 8: 1196-1207

[45] Swiegers JH, Bartowsky EJ, Henschke PA, Pretorius IS. 2005. Yeast and bacterial modulation of wine aroma and flavour. Australian Journal of Grape and Wine Research 11: 139-173