威宁豆酱中主酵菌株的分离与鉴定
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摘要
以贵州省威宁地区传统方法制作的酱曲为试材,通过分离纯化初步得到22株细菌、10株真菌。将分离纯化的菌株分别接种于豆酱中进行纯种发酵,通过感官评价筛选出主要的发酵菌株,通过形态观察、分子生物学技术及VITEK 2全自动微生物鉴定系统对其进行鉴定,并对其发酵豆酱的理化指标进行测定。结果表明,菌株DX-9和DZ-3发酵豆酱的感官评分最高,分别为94分、90分,且与传统威宁豆酱风味相似,确定为主要发酵菌株。鉴定菌株DX-9为枯草芽孢杆菌(Bacillus subtilis),菌株DZ-3为异常威克汉姆酵母(Wickerhamomyces anomalus)。两菌株发酵豆酱的理化指标符合相关国标要求,且菌株DX-9发酵豆酱优于菌株DZ-3。
Taking the traditional fermented paste koji from Weining area of Guizhou Province as the test material, 22 strains of bacteria and 10 strains of fungi were obtained by isolation and purification. The isolated and purified strains were inoculated separately in soybean paste for pure strain fermentation, the major fermentation strains were screened by sensory evaluation and identified by morphological observation, molecular biological technique and VITEK2 automatic microbial identification system, and the physicochemical indexes of fermented soybean paste were determined. The results showed that the sensory scores of soybean paste fermented by strain DX-9 and DZ-3 were the highest, which was 94 and 90, respectively, and the flavor was similar to that of the traditional Weining soybean paste, which was confirmed as the main fermentation strains. The strain DX-9 was identified as Bacillus subtilis, and the strain DZ-3 was identified as Wickerhamomyces anomalus. The physicochemical indexes of soybean paste fermented by two strains were accorded with the requirements of relevant national standard, and the soybean paste fermented by strain DX-9 was better than that by strain DZ-3.
Taking the traditional fermented paste koji from Weining area of Guizhou Province as the test material, 22 strains of bacteria and 10 strains of fungi were obtained by isolation and purification. The isolated and purified strains were inoculated separately in soybean paste for pure strain fermentation, the major fermentation strains were screened by sensory evaluation and identified by morphological observation, molecular biological technique and VITEK2 automatic microbial identification system, and the physicochemical indexes of fermented soybean paste were determined. The results showed that the sensory scores of soybean paste fermented by strain DX-9 and DZ-3 were the highest, which was 94 and 90, respectively, and the flavor was similar to that of the traditional Weining soybean paste, which was confirmed as the main fermentation strains. The strain DX-9 was identified as Bacillus subtilis, and the strain DZ-3 was identified as Wickerhamomyces anomalus. The physicochemical indexes of soybean paste fermented by two strains were accorded with the requirements of relevant national standard, and the soybean paste fermented by strain DX-9 was better than that by strain DZ-3.
引文
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[20]SUN X,LYU G,LUAN Y,et al.Analyses of microbial community of naturally homemade soybean pastes in Liaoning province of China by Illumina Miseq Sequencing[J].Food Res Int,2018,111(9):50-57.
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[23]JEOON A R,LEE J H,MAH J H.Biogenic amine formation and bacterial contribution in Cheonggukjang,a Korean traditional fermented soybean food[J].LWT-Food Sci Technol,2018,92(6):282-289.
[24]黎瑞珍,符素芳,苏小妹,等.儿童酱油中氨基酸态氮含量的测定[J].食品安全导刊,2018(27):120-121.
[25]CHEN Z Y,FENG Y Z,CUI C,et al.Effects of kojimaking with mixed strains on physicochemical and sensory properties of Chinese-type soy sauce[J].J Sci Food Agr,2015,95(10):2145-2154.
[2]贡汉坤.传统豆酱自然发酵的动态分析及人工接种多菌种发酵研究[D].无锡:江南大学,2004.
[3]武俊瑞,王晓蕊,唐筱扬,等.辽宁传统发酵豆酱中乳酸菌及酵母菌分离鉴定[J].食品科学,2015,36(9):78-83.
[4]JUNG K O,PARK S Y,PARK K Y.Longer aging time increases the anticancer and antimetastatic properties of Doenjang[J].Nutrition,2006,22(5):539-545.
[5]ZHAO J,DAI X,LIU X,et al.Changes in microbial community during Chinese traditional soybean paste fermentation[J].Int J Food Sci Tech,2010,44(12):2526-2530.
[6]KWON D Y,HONG S M,AHN I S,et al.Isoflavonoids and peptides from meju,long-term fermented soybeans,increase insulin sensitivity and exert insulinotropic effects in vitro[J].Nutrition,2011,27(2):244-252.
[7]AMARAL C,TOLOI M R T,VASCONCELOS L D,et al.The role of soybean extracts and isoflavones in hormone-dependent breast cancer:aromatase activity and biological effects[J].Food Funct,2017,8(9):3064-3074.
[8]张巧云,孟祥晨.几种发酵豆酱的微生物组成及理化性质分析[J].食品科技,2013,38(8):266-271.
[9]高秀芝,易欣欣,刘慧,等.东北传统豆酱发酵过程中微生物的多样性[J].生物技术通报,2016,32(4):251-255.
[10]樊敏,胡萍,陈仕梯,等.贵州大方豆酱中优势细菌种群分析与分离鉴定[J].中国酿造,2016,35(12):25-30.
[11]张颖,乌日娜,孙慧君,等.豆酱不同发酵阶段细菌群落多样性及动态变化分析[J].食品科学,2017,38(14):30-35.
[12]KIM T W,LEE J H,KIM S E,et al.Analysis of microbial communities in doenjang,a Korean fermented soybean paste,using nested PCR-denaturing gradient gel electrophoresis[J].Int J Food Microbiol,2009,131(2-3):265-271.
[13]NAM Y D,LEE S Y,LIM S I.Microbial community analysis of Korean soybean pastes by next-generation sequencing[J].Int J Food Microbiol,2012,155(1-2):36-42.
[14]魏景超.真菌鉴定手册[M].上海:上海科学技术出版社,1979:102-105.
[15]R E布坎南,N E吉布斯.伯杰氏细菌手册[M].北京:科学出版社,1994:241-287.
[16]程昌泽,吴拥军,龙菊,等.腐乳毛霉高产蛋白酶菌株的分离与鉴定[J].山地农业生物学报,2008(2):127-133.
[17]贾东旭,吴拥军,李耀中,等.细菌型豆豉发酵芽孢杆菌的筛选与鉴定[J].食品科学,2009,30(5):217-221.
[18]柴洋洋,葛菁萍,宋刚,等.传统发酵豆酱中酵母菌的分离、筛选及功能酵母的鉴定[J].中国食品学报,2013,13(3):183-188.
[19]王新,白琴琴,金清.朝鲜族传统大酱中酵母菌的分离及其产酶特性分析[J].食品工业,2015,36(5):166-169.
[20]SUN X,LYU G,LUAN Y,et al.Analyses of microbial community of naturally homemade soybean pastes in Liaoning province of China by Illumina Miseq Sequencing[J].Food Res Int,2018,111(9):50-57.
[21]KIM T W,LEE J H,PARK M H,et al.Analysis of bacterial and fungal communities in Japanese-and Chinese-fermented soybean pastes using nested PCR-DGGE[J].Curr Microbiol,2010,60(5):315-320.
[22]唐筱扬.自然发酵豆酱酱醅微生物多样性分析[D].沈阳:沈阳农业大学,2017.
[23]JEOON A R,LEE J H,MAH J H.Biogenic amine formation and bacterial contribution in Cheonggukjang,a Korean traditional fermented soybean food[J].LWT-Food Sci Technol,2018,92(6):282-289.
[24]黎瑞珍,符素芳,苏小妹,等.儿童酱油中氨基酸态氮含量的测定[J].食品安全导刊,2018(27):120-121.
[25]CHEN Z Y,FENG Y Z,CUI C,et al.Effects of kojimaking with mixed strains on physicochemical and sensory properties of Chinese-type soy sauce[J].J Sci Food Agr,2015,95(10):2145-2154.