在啤酒工业中,啤酒酵母的絮凝特性是决定啤酒高质量生产的重要因素之一。下面啤酒酵母菌株L-1应用于大生产存在酵母悬浮稳定期较短等问题,导致发酵不彻底,影响了最后的啤酒风味质量。基于此,该研究以菌株L-1为出发菌株,进行常压室温等离子体诱变并对诱变菌株进行初筛和二轮复筛,获得了絮凝性减弱且遗传稳定性能较良好的3株目的菌株179、293和361,其絮凝能力分别降低了6.67％、6.67％和5.56％,死亡率分别降低了31.21％、34.57％和16.93％,对应的发酵液中乙醛含量分别降低了19.53％、19.17％和15.83％,同时其消耗碳源、α-氨基氮、产酒精能力等指标均有所提高,主要高级醇含量无明显变化。这些菌株的获得在啤酒工业生产中具有一定的实际应用潜力。
刘小航
,
阮玉磊
,
宿萌
,
付晓芬
,
贺秀丽
,
郭立芸
,
武晓乐
,
郭学武
,
肖冬光
,
陈叶福
. 弱絮凝性下面啤酒酵母菌株的选育[J]. 食品与发酵工业, 2022
, 48(15)
: 47
-54
.
DOI: 10.13995/j.cnki.11-1802/ts.030107
In beer industry, the flocculation properties of Saccharomyces pastorianus is one of the important factors affecting the production of beer. In the application of Saccharomyces pastorianus L-1 in large-scale production, problems such as short yeast suspension stability period and high mortality rate of reused yeast mud existed, resulting in incomplete fermentation, thus affecting the reduction of undesirable flavor substances and the final beer flavor quality. Therefore, L-1 was used as the starting strain to perform atmospheric and room temperature plasma (ARTP) mutagenesis and the mutagenic strains were screened for the first and second screening. Three target strains 179, 293, and 361 with weakened flocculation and good genetic stability were obtained. The flocculation capacity was reduced by 6.67%, 6.67%, and 5.56% respectively, the mortality rate was reduced by 31.21%, 34.57%, and 16.93% respectively, and the corresponding acetaldehyde content in the fermentation broth was reduced by 19.53%, 19.17%, and 15.83%. Indexes such as carbon source consumption, α-amino nitrogen, and alcohol production capacity had all been improved, and the content of major higher alcohols has not changed significantly. The acquisition of these strains has certain practical application potential in the production of beer industry.
[1] STEWART G.Yeast flocculation:Sedimentation and flotation[J].Fermentation, 2018, 4(2):28.
[2] 崔云前, 袭祥雨, 吉春晖, 等.啤酒酵母絮凝影响因素及改善途径的研究概况[J].食品工业, 2020, 41(3):246-250.
CUI Y Q, XI X Y, JI C H, et al.Research on influencing factors and improving ways of beer yeast flocculation[J].The Food Industry, 2020, 41(3):246-250.
[3] 陆健, 商曰玲, 孙军勇, 等.啤酒酵母提前絮凝的研究进展[J].食品与生物技术学报, 2015, 34(5):449-455.
LU J, SHANG Y L, SUN J Y, et al.Research progress of premature yeast flocculation in beer[J].Journal of Food Science and Biotechnology, 2015, 34 (5):449-455.
[4] SOARES E V.Flocculation in Saccharomyces cerevisiae:A review[J].Journal of Applied Microbiology, 2011, 110(1):1-18.
[5] ZHOU X F, SUO J Y, LIU C F, et al.Genome comparison of three lager yeasts reveals key genes affecting yeast flocculation during beer fermentation[J].FEMS Yeast Research, 2021, 21(4):4.
[6] 张建早, 曹健, 张彦青, 等.絮凝性适中优良啤酒酵母的选育[J].酿酒科技, 2013, (2):57-60.
ZHANG J Z, CAO J, ZHANG Y Q, et al.Screening of fine barm strains with good flocculability[J].Liquor-Making Science & Technology, 2013, (2):57-60.
[7] VAN MULDERS S E, CHRISTIANEN E, SAERENS S M G, et al.Phenotypic diversity of Flo protein family-mediated adhesion in Saccharomyces cerevisiae[J].FEMS Yeast Research, 2009, 9(2):178-190.
[8] DEGREIF D, DE ROND T, BERTL A, et al.Lipid engineering reveals regulatory roles for membrane fluidity in yeast flocculation and oxygen-limited growth[J].Metabolic Engineering, 2017, 41:46-56.
[9] GOVENDER P, DOMINGO J L, BESTER M C, et al.Controlled expression of the dominant flocculation genes FLO1,FLO5,and FLO11 in Saccharomyces cerevisiae[J].Applied and Environmental Microbiology, 2008, 74(19):6 041-6 052.
[10] KARABÍN M, JELÍNEK L, KOTRBA P, et al.Enhancing the performance of brewing yeasts[J].Biotechnology Advances, 2018, 36(3):691-706.
[11] ZHANG X, ZHANG X F, LI H P, et al.Atmospheric and room temperature plasma (ARTP) as a new powerful mutagenesis tool[J].Applied Microbiology and Biotechnology, 2014, 98(12):5 387-5 396.
[12] 徐欢欢, 张红兵, 李会宣, 等.常压室温等离子体技术在微生物诱变中的应用进展[J].生物技术进展, 2020, 10(4):358-362.
XU H H, ZHANG H B, LI H X, et al.Application progress of atmospheric and room temperature plasma technology in microbial mutagenesis[J].Current Biotechnology, 2020, 10 (4):358-362.
[13] WANG L, LI B, WANG S P, et al.Improving multiple stress-tolerance of a flocculating industrial Saccharomyces cerevisiae strain by random mutagenesis and hybridization[J].Process Biochemistry, 2021, 102:275-285.
[14] 冯鹏鹏, 周钰涵, 高杏, 等.低产高级醇工业上面发酵酵母的选育[J].食品与发酵工业, 2022, 48(5):23-28.
FENG P P, ZHOU Y H, GAO X, et al.Breeding of industrial fermentation yeast with low yield of higher alcohol[J].Food and Fermentation Industries, 2022, 48(5):23-28.
[15] 李崎, 刘春凤.ASBC分析方法[M].北京:中国轻工业出版社,2012.
LI Q, LIU C F.ASBC Analysis Method[M].Beijing:China Light Industry Press, 2012.
[16] 张明芳. 高温敏感型啤酒酵母的选育及其自溶研究[D].无锡:江南大学, 2020.
ZHANG M F.Screening of thermosensitive autolytic mutant brewer’s yeast and study on yeast autolysis[D].Wuxi:Jiangnan University, 2020.
[17] ABERNATHY D G, SPEDDING G, STARCHER B.Analysis of protein and total usable nitrogen in beer and wine using a microwell ninhydrin assay[J].Journal of the Institute of Brewing, 2009, 115(2):122-127.
[18] 高莹莹. 低醇葡萄酒酵母菌株的选育[D].天津:天津科技大学, 2020.
GAO Y Y.Breeding of low-ethanol-wine yeasts[D].Tianjin:Tianjin University of Science and Technology, 2020.
[19] MATSUSHIKA A, MORIKAWA H, GOSHIMA T, et al.Effect of fermentation conditions on the flocculation of recombinant Saccharomyces cerevisiae capable of co-fermenting glucose and xylose[J].Applied Biochemistry and Biotechnology, 2014, 174(2):623-631.
