永川毛霉型豆豉在发酵过程中菌群动态变化规律
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摘要
永川豆豉是重庆地区的特色传统发酵食品,具有丰富的营养价值。永川豆豉属于开放式制曲、多菌系发酵,豆豉菌群的动态变化一直以来是研究的热点。实验采用PCR-DGGE技术(变性梯度凝胶电泳)对永川毛霉型豆豉发酵过程中真菌和细菌群落的动态变化进行了研究。结果表明:在真菌水平上,制曲阶段总状毛霉(Mucor racemosus)占绝对优势,同时也有匍枝根霉(Rhizopus stolonifer)等辅助发酵,后发酵阶段有假丝酵母(Candida sp.)等酵母菌的参与。在细菌水平上,制曲阶段乳酸乳球菌属(Lactococcus lactis)生长占优势,拌料后其丰度急剧减少,而枯草芽孢杆菌(Bacillus subtilis)等成为新的优势菌株。发酵过程中也出现了一些杂菌,说明开放式制曲的发酵方式会有引入杂菌的安全隐患。
Yongchuan fermented soya beans are traditional fermented food with rich nutritional value in Chongqing.Yongchuan fermented soya beans belong to open-type koji making and multi-strain fermentation.The dynamic change of flora of fermented soya beans has always been a research hotspot.In this study,PCR-DGGE technology(denaturing gradient gel electrophoresis)is used to study the dynamic changes of fungi and bacterial communities during the fermentation of mucor-type Yongchuan fermented soya beans.The results show that the most dominant fungus is Mucor racemosus during koji-making stage,along with Rhizopus stolonifer and etc..Candida grows vigorously during post-fermentation stage.On the bacterial level, Lactococcus lactis grows predominantly in the koji-making stage,and its abundance decreases sharply after mixing,while Bacillus subtilis becomes new dominant strain.There are also some fungi in the process of fermentation,which indicates that the open-type koji-making fermentation method has the potential safety hazard of introducing sundry fungi.
Yongchuan fermented soya beans are traditional fermented food with rich nutritional value in Chongqing.Yongchuan fermented soya beans belong to open-type koji making and multi-strain fermentation.The dynamic change of flora of fermented soya beans has always been a research hotspot.In this study,PCR-DGGE technology(denaturing gradient gel electrophoresis)is used to study the dynamic changes of fungi and bacterial communities during the fermentation of mucor-type Yongchuan fermented soya beans.The results show that the most dominant fungus is Mucor racemosus during koji-making stage,along with Rhizopus stolonifer and etc..Candida grows vigorously during post-fermentation stage.On the bacterial level, Lactococcus lactis grows predominantly in the koji-making stage,and its abundance decreases sharply after mixing,while Bacillus subtilis becomes new dominant strain.There are also some fungi in the process of fermentation,which indicates that the open-type koji-making fermentation method has the potential safety hazard of introducing sundry fungi.
引文
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[28]索化夷,卢露,吴佳敏,等.永川豆豉在传统发酵过程中基本成分及蛋白酶活性变化[J].食品科学,2011,32(1):177-180.
[29]Noval R M,Burton O T,Wise P,et al.A microbiota signature associated with experimental food allergy promotes allergic sensitization and anaphylaxis[J].Journal of Allergy &Clinical Immunology,2013,131(1):201-212.
[2]徐聪聪.固态发酵豆粕制备大豆抗氧化活性肽的研究[D].广州:暨南大学,2013.
[3]王鹏娇,吴运莉,张敏,等.21种不同产地中药淡豆豉中5种活性成分的对比[J].中国实验方剂学杂志,2014,20(19):64-67.
[4]Oh J H,Suh J W,Kim J Y,et al.Development of a genistein-enriched Doenjang using cornβ-glucosidase[J].Food Science &Biotechnology,2008,17(5):1021-1024.
[5]江侧燕,黎金兰,赵树进,等.富含纤溶酶豆豉冻干粉的消化性质及抗血栓作用[J].食品工业科技,2010(9):334-336.
[6]葛喜珍,王鑫国,徐华州,等.中药淡豆豉有效成分大豆异黄酮调节血脂的研究进展[J].河北中医药学报,2002,17(3):41-43.
[7]胡国军,吴天祥.豆豉营养评价及其粗纤溶酶活性检测[J].中国酿造,2010,29(10):95-99.
[8]邹磊,汪立君,呼晴,等.后发酵过程中乙醇对豆豉抗氧化能力的影响[J].食品与发酵工业,2006(12):28-32.
[9]阚建全,陈宗道,石轶松,等.豆豉非透析类黑精抗氧化和抑制亚硝胺合成的研究[J].营养学报,1999(3):349-352.
[10]Park J S,Park H Y,Kim D H,et al.Ortho-dihydroxyisoflavone derivatives from aged Doenjang(Korean fermented soypaste)and its radical scavenging activity[J].Bioorganic &Medicinal Chemistry Letters,2008(18):5006-5009.
[11]郭瑞华,王和平,焦丽艳,等.豆豉中大豆异黄酮及苷元对四氧嘧啶糖尿病模型小鼠血糖及组织的活性作用[J].现代预防医学,2009,36(20):3919-3921.
[12]杜木英,陈宗道,王光慈,等.豆豉保健功能成分研究进展[J].粮食与油脂,2003(2):9-11.
[13]Jeyaram K,Mohendro S W,Premarani T,et al.Molecular identification of dominant microflora associated with"Hawaijar"-a traditional fermented soybean(Glycine max L.)food of Manipur,India[J].International Journal of Food Microbiology,2008,122(3):259-268.
[14]Florez A B,Mayo B.PCR-DGGE as a tool for characterizing dominant microbial populations in the Spanish blue-veined cabrales cheese[J].International Dairy Journal,2006,16(10):1205-1210.
[15]Fasoli S,Marzotto M,Rizzotti L,et al.Bacterial composition of commercial probiotic products as evaluated by PCR-DGGE analysis[J].International Journal of Food Microbiology,2003,82(1):59-70.
[16]Temmerman R,Scheirlinck I,Huys G,et al.Cultureindependent analysis of probiotic products by denaturing gradient gelelectrophoresis[J].Appl Environ Microbiol,2003,69(1):220-226.
[17]Oberauner L,Zachow C,Lackner S,et al.The ignored diversity:complex bacterial communities in intensive care units revealed by 16Spyrosequencing[J].Sci Rep,2013(3):1413.
[18]Jiang X T,Peng X,Deng G H,et al.Illumina sequencing of16S rRNA tag revealed spatial variations of bacterial communities in a mangrove wetland[J].Microbial Ecology,2013,66(1):96-104.
[19]Wang Y,Sheng H F,He Y,et al.Comparison of the levels of bacterial diversity in freshwater,intertidal wetland,and marine sediments by using millions of Illumina tags[J].Applied &Environmental Microbiology,2012,78(23):8264.
[20]Chen C,Xiang J Y,Hu W,et al.Identification of key microorganisms involved in Douchi fermentation by statistical analysis and their use in an experimental fermentation[J].Journal of Applied Microbiology,2015,119(5):1324-1334.
[21]Chen T,Jiang S,Xiong S,et al.Application of denaturing gradient gel electrophoresis to microbial diversity analysis in Chinese Douchi[J].Journal of the Science of Food &Agriculture,2012,92(10):2171-2176.
[22]Chen T,Wang M,Li S,et al.Molecular identification of microbial community in surface and undersurface douchi during postfermentation[J].Journal of Food Science,2014,79(4):653-658.
[23]索化夷,赵欣,骞宇,等.永川毛霉型豆豉在发酵过程中微生物总量与区系变化规律[J].食品科学,2015,36(19):124-131.
[24]Muyzer G,Waal E C D,Uitterlinden A G.Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reactionamplified genes coding for 16SrRNA[J].Appl Environ Microbiol,1993,59(3):695-700.
[25]Muyzer G,Brinkhoff T,Ulrich N,et al.Denaturing gradient gel electrophoresis(DGGE)in microbial ecology[J].Molecular Microbial Ecology Manual,1998(3):1-27.
[26]Li A J,Yang S F,Li X Y,et al.Microbial population dynamics during aerobic sludge granulation at different organic loading rates[J].Water Research,2008,42(13):3552-3560.
[27]Yuko T H,Sho M.Comparison of 18SrDNA primers for estimating fungal diversity in agricultural soils using polymerase chain reaction-denaturing gradient gel electrophoresis[J].Soil Science &Plant Nutrition,2010,54(5):701-710.
[28]索化夷,卢露,吴佳敏,等.永川豆豉在传统发酵过程中基本成分及蛋白酶活性变化[J].食品科学,2011,32(1):177-180.
[29]Noval R M,Burton O T,Wise P,et al.A microbiota signature associated with experimental food allergy promotes allergic sensitization and anaphylaxis[J].Journal of Allergy &Clinical Immunology,2013,131(1):201-212.