Isolation, identification and characteristics of saccharifying strains for rice wine
LI Zeyang1,2, WU Shihua1,2, LONG Xiufeng1,2*, WU Jun3, YI Yi1,2
1(College of Biological and Chemical Engineering,Guangxi University of Science and Technology, Liuzhou 545006, China) 2(Guangxi Key Laboratory of Green Processing of Sugar Resources Liuzhou 545006, China) 3(Teaching Quality Monitoring and Evaluation Center,Guangxi University of Science and Technology Liuzhou 545006, China)
Abstract: In order to obtain strains with strong ability of starch saccharification in rice wine brewing, spore purification method was used to screen molds from nine kinds of Jiuqu(liquor fermentation starter), and liquefying and saccharifying power were determined to rescreen strains with outstanding saccharification ability. The results indicated that among the 13 molds obtained by primary screening, strains 1M5, 5M2 and 7M1 had high liquefying and saccharifying powers, which were (1 315±46.54) and (1 252±63.78), (1 868±92.74) and (1 556±15.56), (1 929±94.55) and 1 054±21.11 mg/(g·h), respectively. According to morphological and phylogenetic analysis, strains 1M5, 5M2 and 7M1 were identified as Rhizopus microsporus, Aspergillus clavatus and Aspergillus niger, respectively. Moreover, saccharifying rice tests showed that the saccharification rates of strains 1M5, 5M2 and 7M1 were 62.55%, 55.61% and 67.04% respectively. In addition, the flavor compounds of strain 5M2 which exhibited outstanding protease activity were analyzed, and 13 main flavor compounds were detected, among which 3-methylvaleric acid, benzylalcohol, phenylethanol, phenylacetic acid and ethylpentadecanoate showed pleasant fragrance. In summary, strains 1M5, 5M2 and 7M1 have good saccharification performance, showing great application potential in rice wine brewing.
[1] LIANG Z C, LIN X Z, HE Z G, et al.Dynamic changes of total acid and bacterial communities during the traditional fermentation of Hong Qu glutinous rice wine[J].Electronic Journal of Biotechnology, 2020, 43:23-31. [2] LYU X C, WENG X, ZHANG W, et al.Microbial diversity of traditional fermentation starters for Hong Qu glutinous rice wine as determined by PCR-mediated DGGE[J].Food Control, 2012, 28(2):426-434. [3] 贾瑞博, 胡荣康, 周文斌, 等.米曲霉(Aspergillus oryzae FAFU)淀粉酶的分离纯化及其酶学性质研究[J].食品与发酵工业, 2016, 42(11):71-76. JIA R B, HU R K, ZHOU W B, et al.Purification and characterization of amylase produced by Aspergillus oryzae FAFU from rice wine fermentation starter[J].Food and Fermentation Industries, 2016, 42(11):71-76. [4] 苏小军, 熊兴耀, 谭兴和, 等.产生淀粉酶菌株的诱变选育及酶学性质研究[J].食品与机械, 2009,25(2):11-14. SU X J, XIONG X Y, TAN X H, et al.Study on the mutation breeding of raw-starch-hydrolysing amylase-producing strain and the enzyme properties[J].Food & Machinery, 2009,25(2):11-14. [5] 罗惠波, 谢军, 黄治国, 等.纯种根霉麸曲制曲工艺优化研究[J].四川理工学院学报(自然科学版), 2015, 28(5):7-11. LUO H B, XIE J, HUANG Z G, et al.Research on optimization of purebred rhizopus bran koji making technics[J].Journal of Sichuan University of Science & Engineering(Natural Science Edition)[J].2015, 28(5):7-11. [6] JIN Y, LI D Y, AI M, et al.Correlation between volatile profiles and microbial communities:A metabonomic approach to study Jiang- flavor liquor Daqu[J].Food Research International, 2019, 121:422-432. [7] 曹文悦, 李灵灵, 高亚梅, 等.一株糖化真菌T6的分离筛选[J].黑龙江八一农垦大学学报, 2019, 31(2):59-64;71. CAO W Y, LI L L, GAO Y M, et al.Isolation and screening of the saccharifying fungus[J].2019, 31(2):59-64;71. [8] 杨勇. 方便甜酒酿的关键技术的研究[D].南昌:江西农业大学, 2012. YANG Y.Study on the key production technology of instant sweet rice wine[D].Nanchang:Jiangxi Agricultural University, 2012. [9] 龙婷, 余志晟, 汪强, 等.我国传统白酒酿造与工业酒精发酵的比较分析[J].中国酿造, 2018, 37(12):7-11. LONG T, YU Z S, WANG Q, et al.Comparative analysis of traditional Baijiu brewing and industrial alcohol fermentation in China[J].China Brewing, 2018, 37(12):7-11. [10] CHEN B, WU Q, XU Y.Filamentous fungal diversity and community structure associated with the solid state fermentation of Chinese Maotai-flavor liquor[J].International Journal of Food Microbiology, 2014, 179:80-84. [11] ZHENG X W, YAN Z, HAN B, et al.Complex microbiota of a Chinese “Fen” liquor fermentation starter (Fen-Daqu), revealed by culture-dependent and culture-independent methods[J].Food Microbiology, 2012, 31(2):293-300. [12] DU H, WANG X S, ZHANG Y H, et al.Exploring the impacts of raw materials and environments on the microbiota in Chinese Daqu starter[J].International Journal of Food Microbiology, 2019, 297:32-40. [13] YAO L D, JU X, JAMES T Y, et al.Relationship between saccharifying capacity and isolation sources for strains of the Rhizopus arrhizus complex[J].Mycoscience, 2018, 59(5):409-414. [14] 孙晓璐, 王明跃, 张源.白酒大曲产酯化酶犁头霉的分离鉴定及产酶条件优化[J].食品与机械, 2018, 34(10):40-44;123. SUN X L, QANG Y M, ZHANG Y.Isolation and identification of high-yield esterase Absidiain Daqu and optimization of enzyme production conditions[J].Food & Machinery, 2018, 34(10):40-44;123. [15] 刘双平, 谢雅茜, 江正强, 等.嗜热拟青霉糖化酶的性质及其在黄酒酿造中的应用[J].酿酒科技, 2019(6):23-29;34. LIU S P, XIE Y Q, JIANG Z Q, et al.Properties of a glucoamylase (PtGA15) from paecilomyces thermophila and its application in Huangjiu production[J].Liquor-Making Science & Technology, 2019(6):23-29;34. [16] 王和玉, 刘延峰, 张巧玲, 等.1株来源于茅台酒酿造过程宛氏拟青霉MTDF-01的全基因组测序及分析[J].食品科学, 2019, 40(24):185-192. WANG H Y, LIU Y F, ZHANG Q L, et al.Whole-genome sequencing and analysis of paecilomyces variotii MTDF-01, isolated during maotai-flavor liquor brewing[J].Food Science, 2019, 40(24):185-192. [17] 龙茜萍, 王晓丹, 谭静, 等.一株高产糖化酶菌株的筛选与鉴定[J].酿酒科技, 2013(8):7-9. LONG X P, WANG X D, TAN J, et al.Screening and identification of a high-yield glucoamylase-producing strain[J].Liquor-Making Science & Technology, 2013(8):7-9. [18] 谭映月, 胡萍, 谢和.我国白酒酿造微生物多样性的研究现状及展望[J].酿酒科技, 2011(11):100-105. TAN Y Y, HU P, XIE H.Present research status and prospects of the diversity of liquor-making microbes in China[J].Liquor-Making Science & Technology, 2011(11):100-105. [19] 王晓燕, 王蓉福, 张昊宇, 等.青稞酒曲中糖化菌的筛选鉴定及降低青稞淀粉工艺优化[J].食品与发酵工业, 2020, 46(5):160-165. WANG X Y, WANG F R, ZHANG H Y, et al.Screening and identification of saccharifying strains in the highland barley koji and process optimization for degrading barley starch[J].Food and Fermentation Industries, 2020, 46(5):160-165. [20] 程伟, 张杰, 潘天全, 等.安琪生香活性干酵母在固态小曲清香型白酒酿造生产中的应用[J].酿酒科技, 2018(7):83-88;91. CHENG W, ZHANG J, PAN T Q, et al.Application of angel aroma-producing active dry yeast in the production of qingxiang Xiaoqu Baijiu by solid-state fermentation[J].Liquor-Making Science & Technology, 2018(7):83-88;91. [21] 韩英, 赵恒山, 田宇敏, 等.基于风味和产酶性能的霉菌M_2的筛选及制曲工艺优化[J].酿酒科技, 2019(9):27-34. HAN Y, ZHAO H S, TIAN Y M, et al.Screening of mold strain M2 based on flavor and enzyme-producing performance and optimization of its Daqu-making process[J].Liquor-Making Science & Technology, 2019(9):27-34. [22] 中华人民共和国工业和信息化部. QB/T 4257—2011 酿酒大曲通用分析方法[S].北京:中国标准出版社, 2011. Ministry of Industry and Information Technology of the People's Republic of China.QB/T 4257—2011 General methods of analysis for Daqu[S].Beijing:China Standards Press, 2011. [23] 魏景超. 真菌鉴定手册[M].上海:上海科学技术出版社, 1979. WEI J C.Manual of Fungal Identification[M].Shanghai:Shanghai Scientific & Technical Publishers, 1979. [24] 中华人民共和国国家质量监督检验检疫局, 中国国家标准化管理委员会.GB/T 23527—2009 蛋白酶制剂[S].北京:中国标准出版社, 2009. State Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, China National Standardization Administration Committee.GB/T 23527—2009 Protease preparations[S].Beijing:China Standards Press, 2009. [25] 黄家驷, 刘盾, 陈玄阳, 等.浓香型高温酒曲中蛋白酶产生菌的筛选与鉴定[J].安徽农学通报, 2017, 23(21):29-31. HUANG J S, LIU D, CHEN X Y, et al.Isolation and identification of protease-producing srains from luzhou-flavor liquor starter[J].Anhui Agricultural Science Bulletin, 2017, 23(21):29-31. [26] 王鹏昊, 关统伟, 邓奥宇, 等.酒曲中高产蛋白酶和α-淀粉酶霉菌菌株的筛选与分子鉴定[J].酿酒科技, 2016(6):61-64;71. WANG P H, GUAN T W, DENG A Y, et al.Screening and molecular identification of fungal strains with high-yield of protease and α-amylase[J].Liquor-Making Science & Technology, 2016(6):61-64;71. [27] 梁香, 吴天祥, 敖锐, 等.黄酒优良糖化菌株的筛选及其酶活性能的研究[J].酿酒科技, 2015(3):12-15. LIANG X, WU T X, AO R, et al.Screening of an excellent saccharifying mold strain for yellow rice wine and study of its enzyme activity[J].Liquor-Making Science & Technology, 2015(3):12-15. [28] CHANCHAROONPONG C, HSIEH P H, SHEU S C.Enzyme production and growth of Aspergillus oryzae S.on soybean koji fermentation[J].APCBEE Procedia, 2012, 2:57-61. [29] 耿晓杰, 张玉红, 薛洁, 等.西藏青稞小曲中优良微生物的筛选与鉴定[J].食品与发酵工业, 2019, 45(15):66-73. GENG X J, ZHANG Y H, XUE J, et al.Screening and identification of excellent microorganisms in Tibetan highland barley Xiaoqu[J].Food and Fermentation Industries, 2019, 45(15):66-73. [30] 吴轩德. 筛选多菌种发酵提升米酒特征香气物质含量的研究[D].广州:华南理工大学, 2018. WU X D.Study on improving the content of characteristic aroma substances in rice wine by screening multi-strain fermentation[D].Guangzhou:South China University of Technology, 2018.
2021, Vol. 47 
