Chen Chong, Zheng Zhaojun, Wang Yue, Ji Xiaoying, Li Dongliang, Li Pinhe, Liu Yuanfa
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, National Engineering Research Center for Functional Food, National Engineering Laboratory for Cereal Fermentation Technology, Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan University, 1800 Lihu Road, Wuxi 214122, Jiangsu, People's Republic of China.
Cigar Fermentation Technology Key Laboratory of China Tobacco (China Tobacco Sichuan Industrial Co., Ltd.), China Tobacco Technology Innovation Center for Cigar, Chengdu 610000, Sichuan, People's Republic of China.
Food Res Int. 2025 Jan;199:115393. doi: 10.1016/j.foodres.2024.115393. Epub 2024 Nov 24.
Ester-producing strains are of great importance for enhancing the quality and flavor profiles of alcoholic beverages. However, traditional methods for screening ester-producing strains are labor-intensive and time-consuming, significantly impeding the development of alcoholic beverages industry. In this study, we selected five brands of Jiuqu to cultivate within different media, results showed that XB (Jiuqu) incubated with malt extract medium possessed the highest ester-producing capability, with the identification of 27 esters at the concentration of 31.44 ± 2.17 mM. Subsequently, fluorescence-activated cell sorting (FACS) was used to screen and isolate the living fungal cells which accounted for 42.80 % of total cells, followed by cultivation utilizing a culturomics approach. High-throughput screening (HTS) assays using 4-Methylumbelliferyl acetate (4-MA) were utilized to evaluate the ester-production potentials of 960 selected strains. The top10 highest ester-producing strains were sequenced, and all were identified as Saccharomyces cerevisiae, Cyberlindnera fabianii, and Wickerhamomyces anomalus. Eventually, three microbial strains were co-incubated with rice wine starter to improve the nutritional and flavor properties of glutinous rice wine. Compared to the control group, Cyberlindnera fabianii could increase reducing sugar content, up to 0.33 ± 0.01 g/mL, and significantly decrease the concentration of bitter amino acids by 55.83 %, resulting in a final concentration of 32.29 ± 1.51 mg/L. Furthermore, the glutinous rice wine with Cyberlindnera fabianii showed 38 distinct ester compounds at the content of 16.22 ± 0.51 mM, which was superior to the control group (30 ester compounds at the concentration of 11.89 ± 1.39 mM). The diversity and concentrations of flavor components, such as alcohols, aldehydes, and ketones, were also enhanced. Our findings would contribute to advancing the rapid screening of ester-producing strains, as well as providing a theoretical basis for improving the quality of glutinous rice wine.
产酯菌株对于提升酒精饮料的品质和风味特征至关重要。然而，传统的筛选产酯菌株的方法 labor-intensive 且耗时，严重阻碍了酒精饮料行业的发展。在本研究中，我们选取了五个品牌的酒曲在不同培养基中培养，结果显示，在麦芽提取物培养基中培养的 XB（酒曲）产酯能力最高，鉴定出 27 种酯，浓度为 31.44 ± 2.17 mM。随后，使用荧光激活细胞分选（FACS）筛选并分离出占总细胞 42.80%的活真菌细胞，接着采用 culturomics 方法进行培养。使用乙酸 4-甲基伞形酮酯（4-MA）的高通量筛选（HTS）分析用于评估 960 株选定菌株的产酯潜力。对产酯能力最高的前 10 株菌株进行测序，所有菌株均被鉴定为酿酒酵母、法比安有孢圆酵母和异常威克汉姆酵母。最终，将三株微生物菌株与米酒酒曲共同培养以改善糯米酒的营养和风味特性。与对照组相比，法比安有孢圆酵母可使还原糖含量增加至 0.33 ± 0.01 g/mL，并显著降低苦味氨基酸浓度 55.83%，最终浓度为 32.29 ± 1.51 mg/L。此外，含有法比安有孢圆酵母的糯米酒显示出含量为 16.22 ± 0.51 mM 的 38 种不同酯类化合物，优于对照组（浓度为 11.89 ± 1.39 mM 的 30 种酯类化合物）。醇类、醛类和酮类等风味成分的多样性和浓度也有所提高。我们的研究结果将有助于推进产酯菌株的快速筛选，并为提高糯米酒质量提供理论依据。
