CAI Weiqi, ZHONG Wujie, ZHOU Jiajian, et al. Isolation, Identification, Optimization of Fermentation Conditions and Application of β-Phenylethanol-producing Yeast from Natural Soy Sauce Mash[J]. Science and Technology of Food Industry, 2023, 44(17): 149−157. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022110011.
Citation: CAI Weiqi, ZHONG Wujie, ZHOU Jiajian, et al. Isolation, Identification, Optimization of Fermentation Conditions and Application of β-Phenylethanol-producing Yeast from Natural Soy Sauce Mash[J]. Science and Technology of Food Industry, 2023, 44(17): 149−157. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022110011.

Isolation, Identification, Optimization of Fermentation Conditions and Application of β-Phenylethanol-producing Yeast from Natural Soy Sauce Mash

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  • Received Date: November 01, 2022
  • Available Online: July 02, 2023
  • To improve the quality of soy sauce, this paper presented a study on the production of β-phenylethanol by Zygosaccharomyces rouxii and its application in the brewing of high-salt liquid-state fermented soy sauce. The β-phenylethanol-producing yeast strain J13, screened from natural sauce mash, was identified by morphological observation, physiological and biochemical results and molecular biological methods. The biological characteristics of Z. rouxii 13 through salt tolerance, pH tolerance and temperature tolerance analysis were studied. A series of single-factor and orthogonal tests were conducted to get the optimized medium composition and fermentation conditions for this strain. Z. rouxii 13 was then cultured under the optimal fermentation conditions to obtained flavor liquid and applied to the brewing process of high-salt liquid-state fermented soy sauce. The results showed that J13 was identified as Zygosaccharomyces rouxii, named Z. rouxii 13 accordingly. Z. rouxii 13 could successfully grow under high salinity (18% NaCl) conditions. The optimum pH was 4~8 and the optimum temperature was 25~35 ℃. Under the optimal fermentation conditions: L-phenylalanine concentration of 4 g/L, glucose concentration of 30 g/L, peptone concentration of 10 g/L, magnesium sulfate concentration of 0.5 g/L, dipotassium hydrogen phosphate concentration of 5 g/L, NaCl concentration of 50 g/L, pH6.0, and temperature of 28 ℃, the highest content with β-phenylethanol of 1.40 g/L was obtained. The flavor liquid was applied to soy sauce brewing and the finished products were obtained. The sensory evaluation showed that the flavor liquid group had more prominent soy sauce flavor and fullness, and better umami and overall evaluation compared with the control group that without flavor liquid.
  • [1]
    DESMOND K. The role of microorganisms in soy sauce production[M]. London: Advances in Applied Microbiology, 2019: 45−113.
    [2]
    QI Q, HUANG J, ZHOU R Q, et al. Characterizing microbial community and metabolites of Cantonese soy sauce[J]. Food Bioscience,2020,40:100872.
    [3]
    刘晓艳, 叶月华, 钱敏, 等. 大型发酵酱油酿造过程中风味物质的动态变化分析[J]. 食品科学,2021,42(12):242−247. [LIU X Y, YE Y H, QIAN M, et al. Analysis of dynamic changes of flavor components during Large-Scale brewing of soy sauce[J]. Food Science,2021,42(12):242−247. doi: 10.7506/spkx1002-6630-20200514-167

    LIU X Y, YE Y H, QIAN M, et al. Analysis of dynamic changes of flavor components during Large-Scale brewing of soy sauce[J]. Food Science, 2021, 42(12): 242-247. doi: 10.7506/spkx1002-6630-20200514-167
    [4]
    DIEZ-SIMON C, EICHELSHEIM C, MUMM R, et al. Chemical and sensory characteristics of soy sauce: A review[J]. Journal of Agricultural and Food Chemistry,2020,68(42):11612−11630. doi: 10.1021/acs.jafc.0c04274
    [5]
    DEVANTHI P V P, GKATZIONIS K. Soy sauce fermentation: Microorganisms, aroma formation, and process modification[J]. Food Research International,2019,120:364−374. doi: 10.1016/j.foodres.2019.03.010
    [6]
    FENG Y Z, CUI C, ZHAO H F, et al. Effect of koji fermentation on generation of volatile compounds in soy sauce production[J]. International Journal of Food Science & Technology,2013,48(3):609−619.
    [7]
    葛金鑫, 李永凯, 曾斌. 酱油的风味物质[J]. 中国酿造,2019,38(10):16−20. [GE J X, LI Y K, ZENG B. Flavor substances in soy sauce[J]. China Brewing,2019,38(10):16−20. doi: 10.11882/j.issn.0254-5071.2019.10.004

    GE J X, LI Y K, ZENG B. Flavor substances in soy sauce[J]. China Brewing, 2019, 38(10): 16-20. doi: 10.11882/j.issn.0254-5071.2019.10.004
    [8]
    LEE S M, KIM S B, KIM Y S. Determination of key volatile compounds related to Long-Term fermentation of soy sauce[J]. Journal of Food Science,2019,84(10):2758−2776. doi: 10.1111/1750-3841.14771
    [9]
    陈杰, 赵莹, 韩舜羽, 等. 17种市售广式酱油中风味物质的检测分析[J]. 中国酿造,2021,40(12):165−170. [CHEN J, ZHAO Y, HAN S Y, et al. Detection and analysis of flavor substances in 17 kinds of commercially available Cantonese soy sauce[J]. China Brewing,2021,40(12):165−170. doi: 10.11882/j.issn.0254-5071.2021.12.030

    CHEN J, ZHAO Y, HAN S Y, et al. Detection and analysis of flavor substances in 17 kinds of commercially available Cantonese soy sauce[J]. China Brewing, 2021, 40(12): 165-170. doi: 10.11882/j.issn.0254-5071.2021.12.030
    [10]
    白佳伟, 陈亮, 周尚庭, 等. 特级高盐稀态酿造酱油中关键香气物质的分析[J]. 中国酿造,2019,38(11):179−185. [BAI J W, CHEN L, ZHOU S T, et al. Analysis of key odor compounds in super grade soy sauce with high-salt and diluted-state fermentation[J]. China Brewing,2019,38(11):179−185. doi: 10.11882/j.issn.0254-5071.2019.11.036

    BAI J W, CHEN L, ZHOU S T, et al. Analysis of key odor compounds in super grade soy sauce with high-salt and diluted-state fermentation[J]. China Brewing, 2019, 38(11): 179-185. doi: 10.11882/j.issn.0254-5071.2019.11.036
    [11]
    冯云子. 高盐稀态酱油关键香气物质的变化规律及形成机理的研究[D]. 广州: 华南理工大学, 2015

    FENG Y Z. The evolution and formation mechanism of keyaroma compounds during the process of high-salt liquid fermentation soy sauce[D]. Guangzhou: South China University of Technology, 2015.
    [12]
    STEINHAUS P, SCHIEBERLE P. Characterization of the key aroma compounds in soy sauce using approaches of molecular sensory science[J]. Journal of Agricultural and Food Chemistry,2007,55(15):6262−6269. doi: 10.1021/jf0709092
    [13]
    WANG X J, GUO M Y, SONG H L, et al. Characterization of key aroma compounds in traditional Chinese soy sauce through the molecular sensory science technique[J]. LWT-Food Science and Technology,2020,128:1096−1127.
    [14]
    朱新贵, 李学伟, 曾小波. 典型广式酱油与日式酱油的风味物质差异研究[J]. 中国酿造,2016,35(7):30−35. [ZHU X G, LI X W, ZENG X B. Difference of flavor compounds in Cantonese-style soy sauce and Japanese-style soy sauce[J]. China Brewing,2016,35(7):30−35. doi: 10.11882/j.issn.0254-5071.2016.07.007

    ZHU X G, LI X W, ZENG X B. Difference of flavor compounds in Cantonese-style soy sauce and Japanese-style soy sauce[J]. China Brewing, 2016, 35(7): 30-35. doi: 10.11882/j.issn.0254-5071.2016.07.007
    [15]
    彭东, 蒋雪薇, 陈幽, 等. 高盐稀态酱醪中耐盐生香酵母的筛选及生香特性研究[J]. 食品与发酵工业,2020,46(13):76−84. [PENG D, JIANG X W, CHEN Y, et al. Screening and aroma-producing characteristics of salt-tolerant aroma-producing yeasts from high-salt liquid-state Moromi[J]. Food and Fermentation Industries,2020,46(13):76−84.

    PENG D, JIANG X W, CHEN Y, et al. Screening and aroma-producing characteristics of salt-tolerant aroma-producing yeasts from high-salt liquid-state Moromi[J]. Food and Fermentation Industries, 2020, 46(13): 76-84.
    [16]
    YAO S J, ZHOU R Q, JIN Y, et al. Effect of co-culture with Tetragenococcus halophilus on the physiological characterization and transcription profiling of Zygosaccharomyces rouxii[J]. Food Research International,2019,121:348−358. doi: 10.1016/j.foodres.2019.03.053
    [17]
    阮志强, 董玺梅, 蒋雪薇, 等. 高盐稀态酱油发酵优势真菌与风味物质相关性分析[J]. 食品科学,2021,43(10):1−11. [RUAN Z Q, DONG X M, JIANG X W, et al. Study on the correlation between dominant fungi and the variation of flavor compounds in high-salt liquid-state soy sauce fermentation[J]. Food Science,2021,43(10):1−11. doi: 10.7506/spkx1002-6630-20200514-165

    RUAN Z Q, DONG X M, JIANG X W, et al. Study on the correlation between dominant fungi and the variation of flavor compounds in high-salt liquid-state soy sauce fermentation[J]. Food Science, 2021, 43(10): 1-11. doi: 10.7506/spkx1002-6630-20200514-165
    [18]
    王靖雯, 赵谋明, 陈涛, 等. 高盐稀态酱醪中功能性酵母菌的筛选、鉴定及发酵特性[J]. 食品科学, 2020: 1−11

    WANG J W, ZHAO M M, CHEN T, et al. Screening, identification and fermentation characteristics of functional yeast from high-salt liquid-state moromi[J]. Food Science, 2020: 1−11.
    [19]
    马荣山, 王姗姗. 酱醪中耐高温耐盐酵母菌分离鉴定[J]. 食品工业科技,2013,34(14):191−196. [MA R S, WANG S S. Isolation, purification and identification of high temperature halophilic yeast from soy sauce Mash[J]. Science and Technology of Food Industry,2013,34(14):191−196.

    MA R S, WANG S S. Isolation, purification and identification of high temperature halophilic yeast from soy sauce Mash[J]. Science and Technology of Food Industry, 2013, 34: 191-196.
    [20]
    刘滢, 朱新贵, 李学伟. 酱油增香用耐盐酵母的分离及生长特性分析[J]. 中国酿造,2012,31(10):109−112. [LIU Y, ZHU X G, LI X W. Isolation and growth characteristics of the salt-tolerant yeast for soy sauce flavor improvement[J]. China Brewing,2012,31(10):109−112. doi: 10.3969/j.issn.0254-5071.2012.10.030

    LIU Y, ZHU X G, LI X W. Isolation and growth characteristics of the salt-tolerant yeast for soy sauce flavor improvement[J]. China Brewing, 2012, 31: 109-112. doi: 10.3969/j.issn.0254-5071.2012.10.030
    [21]
    巴尼特, 佩恩, 亚罗. 酵母菌的特征与鉴定手册[M]. 青岛: 青岛海洋大学出版社, 1991: 448

    BARNETT, PENN, YARROW. Yeasts: characteristics and identification[M]. Qingdao: Qingdao Ocean University Press, 1991: 448.
    [22]
    刘慧. 现代食品微生物学实验技术[M]. 北京: 中国轻工业出版社, 2017

    LIU H. Modern experiment technology of food microbiology[M]. 北京: China Light Industry Press, 2017.
    [23]
    SAITOU N, NEI M. The neighbor-joining method: A new method for reconstructing phylogenetic trees[J]. Molecular Biology and Evolution,1987,4(4):406−425.
    [24]
    许岱, 范光森, 富志磊, 等. 一株高产β-苯乙醇酵母菌的筛选、鉴定及其发酵条件优化[J]. 食品工业科技,2017,38(5):151−158. [XU D, FAN G S, FU Z L, et al. Screening of a high-yield β-phenylethanol yeast and optimization of its cultural conditions[J]. Science and Technology of Food Industry,2017,38(5):151−158.

    XU D, FAN G S, FU Z L, et al. Screening of a high-yield β-phenylethanol yeast and optimization of its cultural conditions[J]. Science and Technology of Food Industry, 2017, 38(5): 151-158.
    [25]
    史荣超, 刘洒洒, 侯阳阳, 等. 梅奇酵母转化l-苯丙氨酸合成2-苯乙醇发酵条件优化[J]. 食品与发酵工业,2021,47(15):22−28. [SHI R C, LIU S S, HOU Y Y, et al. Optimized bio-conversion of L-phenylalanine to 2-phenylethanol in Metschnikowia sp

    J]. Food and Fermentation Industries,2021,47(15):22−28.
    [26]
    FAN G S, CHENG L J, FU Z L, et al. Screening of yeasts isolated from Baijiu environments for 2-phenylethanol production and optimization of production conditions[J]. 3 Biotech,2020,10(6):326−332.
    [27]
    高献礼. 高盐稀态酱油在发酵和巴氏杀菌过程中风味物质形成和变化的研究[D]. 广州: 华南理工大学, 2010

    GAO X L. Study on the formation and changes of flavor compounds in high-salt and diluted-state soy sauce during fermentation and pasteurization[D]. Guangzhou: South China University of Technology, 2010.
    [28]
    荣绍丰, 伍进, 管世敏, 等. 生物合成天然2-苯乙醇细菌的筛选及合成途径分析[J]. 食品科学,2021,42(16):69−75. [RONG S F, WU J, GUAN S M, et al. Isolation of 2-Phenylethanol-Synthesizing bacteria and synthetic pathway analysis[J]. Food Science,2021,42(16):69−75. doi: 10.7506/spkx1002-6630-20200515-174

    RONG S F, WU J, GUAN S M, et al. Isolation of 2-Phenylethanol-Synthesizing bacteria and synthetic pathway analysis[J]. Food Science, 2021, 42(16): 69-75. doi: 10.7506/spkx1002-6630-20200515-174
    [29]
    IMAMURA M. Descriptive terminology for the sensory evaluation of soy sauce[J]. Journal of Sensory Studies,2016,31(5):393−407. doi: 10.1111/joss.12223
    [30]
    CHERDCHU P, CHAMBERS E, SUWONSICHON T. Sensory lexicon development using trained panelists in thailand and the USA: Soy sauce[J]. Journal of Sensory Studies,2013,28(3):248−255. doi: 10.1111/joss.12041
    [31]
    陈卫平, 涂谨, 熊建华, 等. 红四氮唑在酒精酵母选育中的应用效果研究[J]. 酿酒科技,2003(6):35−37. [CHEN W P, TU J, XIONG J H, et al. Research on application effect of TTC in breeding of alcohol yeast[J]. Liquor-Making Science & Technology,2003(6):35−37. doi: 10.3969/j.issn.1001-9286.2003.06.007

    CHEN W P, TU J, XIONG J H, et al. Research on application effect of TTC in breeding of alcohol yeast[J]. Liquor-Making Science & Technology, 2003, (6): 35-37. doi: 10.3969/j.issn.1001-9286.2003.06.007
    [32]
    王虎玄, 岳田利, 胡仲秋, 等. 陕西浓缩苹果汁中高渗酵母的分离鉴定[J]. 农业机械学报,2015,46(4):246−251. [WANG H X, YUE T L, HU Z Q, et al. Identification of osmotolerant yeast in apple juice concentrate from shaanxi province[J]. Transactions of the Chinese Society for Agricultural Machinery,2015,46(4):246−251. doi: 10.6041/j.issn.1000-1298.2015.04.036

    WANG H X, YUE T L, HU Z Q, et al. Identification of osmotolerant yeast in apple juice concentrate from shaanxi province[J]. Transactions of the Chinese Society for Agricultural Machinery, 2015, 46(4): 246-251. doi: 10.6041/j.issn.1000-1298.2015.04.036
    [33]
    SENG H T, 周斌, 侯莎, 等. 盐分对广式高盐稀态酱油发酵微生物菌群结构的影响[J]. 食品与发酵工业,2022,48(1):45−54. [SENG T H, ZHOU B, HOU S, et al. Effect of salt on the microbial community structure of Cantonese-style high-salt dilute soy sauce fermentation[J]. Food and Fermentation Industries,2022,48(1):45−54.

    SENG T H, ZHOU B, HOU S, et al. Effect of salt on the microbial community structure of Cantonese-style high-salt dilute soy sauce fermentation[J]. Food and Fermentation Industries, 2022, 48(1): 45-54.
    [34]
    GHOSH S, KEBAARA B W, ATKIN A L, et al. Regulation of aromatic alcohol production in candida albicans[J]. Applied and Environmental Microbiology,2008,74(23):7211−7218. doi: 10.1128/AEM.01614-08
    [35]
    赵佳豪, 周韬, 刘爽, 等. 中日两国酱油香味与滋味差异[J]. 食品与发酵工业,2020,46(17):232−236. [ZHAO J H, ZHOU T, LIU S, et al. Differences in the flavor and taste between Chinese and Japanese soy sauce[J]. Food and Fermentation Industries,2020,46(17):232−236.

    ZHAO J H, ZHOU T, LIU S, et al. Differences in the flavor and taste between Chinese and Japanese soy sauce[J]. Food and Fermentation Industries, 2020, 46(17): 232-236.
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