Rapid screening of industrial brewer’s yeast with low acetaldehyde yield by multi-round ARTP mutagenesis
SUN Kecheng1, YIN Hua2, ZHAO Xinrui1,3,4, CHEN Lu2, LI Jianghua1,3,4, HOU Xiaoping2, DU Guocheng1,3,4,5*
1(School of Biotechnology,Jiangnan University,Wuxi 214122,China) 2(State Key Laboratory of Biological Fermentation Engineering of Beer,Tsingtao Brewery Company Limited,Qingdao 266000,China) 3(Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,China) 4(Science Center for Future Foods,Jiangnan University,Wuxi 214122,China) 5(The Key Laboratory of Carbohydrate Chemistry and Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China)
Abstract: Acetaldehyde is a major carbonyl compound in beer which influences the flavor and stability. The metabolism of yeast is the major source of acetaldehyde for beer. In order to select Saccharomyces cerevisiae with low acetaldehyde and superior fermentability, the industrial Lager yeast was mutated by multi-round ARTP. In addition, disulfiram and acetaldehyde with different contents were added for screening and domesticating mutants. Next, the activities of alcohol dehydrogenase and acetaldehyde dehydrogenase were measured in the initially screened strains. Furthermore, the industrial brewer's yeast with low acetaldehyde was obtained which showed few differences in the major aroma substances, fermentability and biological characteristics. The results showed that in the Lager-16 fermented beer the acetaldehyde decreased by 61.63% from 42.03 mg/L to 16.51 mg/L, and the main aroma components, alcohol, original wort, fermentation rate, growth curve and flocculability were no obvious differences with Lager yeast. Therefore, Lager-16 was the most promising strain for industrial lager beer. The results could provide a theoretical basis for further developing industrial brewer’s yeast and industrial production of high-quality beer.
JAEGER S R,WORCH T,PHELPS T,et al.Effects of “craft” vs.“traditional” labels to beer consumers with different flavor preferences:A comprehensive multi-response approach[J].Food Quality and Preference,2021,87:104 043.
[2]
KUCHARCZYK K,ZYLA K,TUSZYNSKI T.Simultaneous optimization of acetaldehyde and DMS concentrations for better sensory quality of beer fermented on an industrial scale[J].Foods,2020,9(8):1 043.
[3]
SHIN K S,LEE J H.Acetaldehyde contents and quality characteristics of commercial alcoholic beverages[J].Food Science and Biotechnology,2019,28(4):1 027-1 036.
[4]
刘春凤, 赵云,李崎,等.低乙醛Lager型啤酒酵母研究进展[J].菌物学报,2018,37(11):1 411-1 423.LIU C F,ZHAO Y,LI Q,et al.Research advances in lager brewer’s yeast with low acetaldehyde production[J].Mycosystema,2018,37(11):1 411-1 423.
[5]
XU X,NIU C T,LIU C F,et al.Unraveling the mechanisms for low-level acetaldehyde production during alcoholic fermentation in Saccharomyces pastorianus lager yeast[J].Journal of Agricultural and Food Chemistry,2019,67(7):2 020-2 027.
[6]
SANCHEZ R G,SOLODOVNIKOVA N,WENDLAND J.Breeding of lager yeast with Saccharomyces cerevisiae improves stress resistance and fermentation performance[J].Yeast,2012,29(8):343-355.
[7]
沈楠. 低产乙醛啤酒酵母的选育[D].无锡:江南大学,2013.SHEN N.Screening of brewer’s yeast of low acetaldehyde production[D].Wuxi:Jiangnan University,2013.
[8]
张媛媛. 低产乙醛啤酒酵母的选育与研究[D].无锡:江南大学,2011.ZHANG Y Y.Screening of brewer’s yeast with low acetaldehyde[D].Wuxi:Jiangnan University,2011.
[9]
王君伟. 啤酒酵母乙醛代谢关键酶及低乙醛菌株快速筛选的研究[D].济南:齐鲁工业大学,2016.WANG J W.Study on the key enzymes of acetaldehyde metabolism of beer yeast and rapid screening of yeast with low acetaldehyde[D].Jinan:Qilu University of Technology,2016.
[10]
曹荣锟, 李佳泰,王金晶,等.大米辅料啤酒中蛋白质疏水性与蛋白质泡沫稳定性的分析[J].食品与发酵工业,2018,44(9):66-70.CAO R K,LI J T,WANG J J,et al.Hydrophobicity and foaming stability analysis of rice beer protein[J].Food and Fermentation Industries,2018,44(9):66-70.
[11]
JIANG G Z,YANG Z M,WANG Y,et al.Enhanced astaxanthin production in yeast via combined mutagenesis and evolution[J].Biochemical Engineering Journal,2020,156:107 519.
[12]
沈楠, 王金晶,刘春凤,等.低产乙醛啤酒酵母的定向驯化筛选[J].食品与发酵工业,2013,39(7):94-97.SHEN N,WANG J J,LIU C F,et al.Screening of brewer’s yeast with low acetaldehyde by directional domestication[J].Food and Fermentation Industries,2013,39(7):94-97.
[13]
孙传伯, 曹猛,陈存武,等.低产乙醛拉格啤酒酵母的选育[J].皖西学院学报,2020,36(5):64-70.SUN C B,CAO M,CHEN C W,et al.Screening of lager’s yeast of low acetaldehyde production[J].Journal of West Anhui University,2020,36(5):64-70.
[14]
崔云前, 王君伟,李红,等.啤酒酵母乙醛代谢关键酶相关性研究[J].中国酿造,2016,35(1):29-33.CUI Y Q,WANG J W,LI H,et al.Correlation of key enzymes of acetaldehyde metabolism in beer yeast[J].China Brewing,2016,35(1):29-33.
[15]
SUNG C K,KIM S M,OH C J,et al.Taraxerone enhances alcohol oxidation via increases of alcohol dehyderogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities and gene expressions[J].Food and Chemical Toxicology,2012,50(7):2 508-2 514.
[16]
郑英训, 郑飞云,朱林江,等.脱胚玉米糁在啤酒酿造中的应用[J].食品与发酵工业,2016,42(10):87-92.ZHENG Y X,ZHENG F Y,ZHU L J,et al.The application of degermed corn grits as adjunct in beer brewing[J].Food and Fermentation Industries,2016,42(10):87-92.
[17]
朱凤娇, 陈叶福,王希彬,等.上面发酵高粱啤酒的工艺研究[J].现代食品科技,2017,33(9):210-216.ZHU F J,CHEN Y F,WANG X B,et al.Study on the brewing process of top-fermented sorghum beer[J].Modern Food Science and Technology,2017,33(9):210-216.
[18]
SALASPURO M.Local acetaldehyde:Its key role in alcohol-related oropharyngeal cancer[J].Visceral Medicine,2020,36(3):167-173.
[19]
VOLCHENKO N N,SAMKOV A A,MALYSHKO V V,et al.Influence of the environmental isotope composition modification on growth and metabolic activity of Rhodococcus and Saccharomyces[J].Biology Bulletin,2020,47(4):339-343.
[20]
KREGIEL D,BERLOWSKA J,SZUBZDA B.Novel permittivity test for determination of yeast surface charge and flocculation abilities[J].Journal of Industrial Microbiology & Biotechnology,2012,39(12):1 881-1 886.
[21]
XU X,WANG J,BAO M,et al.Reverse metabolic engineering in lager yeast:Impact of the NADH/NAD(+) ratio on acetaldehyde production during the brewing process[J].Applied Microbiology and Biotechnology,2019,103(2):869-880.
[22]
STANISLAV K,KABELOVA-FICOVA H,GREGOR T,et al.Preparation of malts for production of special beers[J].Potravinarstvo Slovak Journal of Food Sciences,2017,11(1):441-445.
[23]
王孟祺. 接种量对上面发酵酵母高级醇代谢的影响及机理研究[D].天津:天津科技大学,2019.WANG M Q.The mechanism of the effect of pitching rate on the metabolism of higher alcohols by top-fermenting yeast[D].Tianjin:Tianjin University of Science & Technology,2019.
[24]
PANTELOGLOU A G,SMART K A,COOK D J.Malt-induced premature yeast flocculation:current perspectives[J].Journal of Industrial Microbiology & Biotechnology,2012,39(6):813-822.