自然发酵甜面酱中芽孢杆菌的分离、筛选及鉴定
|
摘要
利用10倍梯度稀释法涂布平板,从成熟的自然发酵甜面酱中分离、筛选出芽孢杆菌,共计得到13株菌株。通过对其进行菌落、显微形态观察、生理生化实验鉴定和16SrDNA片段扩增及序列分析技术分析,最终确定13株菌株中:菌株B2,B4,B5,B6,B7,B8,B11,B12为枯草芽孢杆菌,菌株B3,B10,B13为解淀粉芽孢杆菌,菌株B1为特基拉芽孢杆菌,菌株B9为短小芽孢杆菌,所得菌株为进一步探究其在甜面酱发酵过程中的作用机理奠定了基础。
Ten times gradient dilution plate method is used for the isolation and screening of Bacillus strain from natural fermented sweet bean sauce.Total 13 strains are screened from the sweet bean sauce.The microscopic,colony,identification test of physiology and biochemistry and 16 SrDNA fragment amplification and sequence analysis reveal that strains B2,B4,B5,B6,B7,B8,B11,B12 are identified as Bacillus subtilis;strains B3,B10,B13 are identified as Bacillus amyloliquefaciens,while strain B1 is Bacillus tequilensis and strain B9 is Bacillus pumilus.All the strains can lay a foundation for the further exploration of their action mechanism in the fermentation process of sweet bean sauce.
Ten times gradient dilution plate method is used for the isolation and screening of Bacillus strain from natural fermented sweet bean sauce.Total 13 strains are screened from the sweet bean sauce.The microscopic,colony,identification test of physiology and biochemistry and 16 SrDNA fragment amplification and sequence analysis reveal that strains B2,B4,B5,B6,B7,B8,B11,B12 are identified as Bacillus subtilis;strains B3,B10,B13 are identified as Bacillus amyloliquefaciens,while strain B1 is Bacillus tequilensis and strain B9 is Bacillus pumilus.All the strains can lay a foundation for the further exploration of their action mechanism in the fermentation process of sweet bean sauce.
引文
[1]路永芳.该传统工艺酿造甜面酱生产技术[J].中国调味品,1995(9):28-29.
[2]宋安东.调味品发酵工艺学[M].北京:化学工业出版社,2009:136-137.
[3]孟鸳.甜面酱发酵过程中挥发性成分的研究[D].武汉:湖北工业大学,2011.
[4]李萍萍,邓静,吴华昌,等.甜面酱自然发酵过程中理化指标及酶活动态变化研究[J].食品科技,2013,38(6):290-294.
[5]冷云伟.酱油中米曲霉及制曲工艺的研究[D].无锡:江南大学,2004.
[6]Elena Di Maro,Danilo Ercolini,Salvatore Coppola.Yeast dynamics during spontaneous wine fermentation of the Catalanesca grape[J].International Journal of Food Microbiology,2007,117(2):201-210.
[7]张忠刚.永丰辣酱自然发酵的菌相与成分分析及人工接种发酵工艺研究[D].长沙:湖南农业大学,2007.
[8]曾灿伟.甜面酱酿制过程中的菌相分析及风味成分研究[D].武汉:湖北工业大学,2009.
[9]Zhao Jianxin,Dai Xiaojun,Liu Xiaoming,et al.Comparison of aroma compounds in naturally fermented and inoculated Chinese soybean pastes by GC-MS and GC-Olfactometry analysis[J].Food Control,2011,22(6):1008-1013.
[10]赵建新.传统豆酱发酵过程分析与控制发酵的研究[D].无锡:江南大学,2011.
[11]康旭,曾灿伟,黄红霞,等.酿造工艺对甜面酱风味成分影响研究[J].中国调味品,2011,36(1):58-60.
[12]孟鸳,乔宇,康旭,等.同时蒸馏萃取、溶剂萃取和顶空固相微萃取与GC-MS联用分析甜面酱的挥发性成分[J].中国调味品,2011,36(1):97-100.
[13]Kotiranta A,Lounatmaa K,Haapasalo M.Epidemiology and pathogenesis of Bacillus cereus infections[J].Microbes and Infection,2000(2):189-198.
[14]Abriouwl H,Maqueda M,Galvez A.Inhibition of bacterial growth,enterotoxin production and sporeout growth in strains of Bacillus cereus by bacteriocin AS-48[J].Appl Environ Mierobiol,2002,68(3):1473-1477.
[15]Zhou J S,Shu Q.Studies on translocation,acute oral toxicity and intestinal colonization of potentially probiotic lactic acid bacteria administ[J].Food Chem Toxieol,2000,38(2):153-161.
[16]吉志伟,刘阳,邓静,等.自然发酵甜面酱中一株芽孢杆菌的分离鉴定及益生特性初探[J].中国调味品,2015,40(12):26-30.
[17]Sekiguchi H,Watanabe M,Nakahara T,et al.Succession of bacterial community structure along the Changjiang River determined by denaturing gradient gel electrophoresis and clone library analysis[J].Appl Environ Microbiol,2002,68(10):5142-5150.
[2]宋安东.调味品发酵工艺学[M].北京:化学工业出版社,2009:136-137.
[3]孟鸳.甜面酱发酵过程中挥发性成分的研究[D].武汉:湖北工业大学,2011.
[4]李萍萍,邓静,吴华昌,等.甜面酱自然发酵过程中理化指标及酶活动态变化研究[J].食品科技,2013,38(6):290-294.
[5]冷云伟.酱油中米曲霉及制曲工艺的研究[D].无锡:江南大学,2004.
[6]Elena Di Maro,Danilo Ercolini,Salvatore Coppola.Yeast dynamics during spontaneous wine fermentation of the Catalanesca grape[J].International Journal of Food Microbiology,2007,117(2):201-210.
[7]张忠刚.永丰辣酱自然发酵的菌相与成分分析及人工接种发酵工艺研究[D].长沙:湖南农业大学,2007.
[8]曾灿伟.甜面酱酿制过程中的菌相分析及风味成分研究[D].武汉:湖北工业大学,2009.
[9]Zhao Jianxin,Dai Xiaojun,Liu Xiaoming,et al.Comparison of aroma compounds in naturally fermented and inoculated Chinese soybean pastes by GC-MS and GC-Olfactometry analysis[J].Food Control,2011,22(6):1008-1013.
[10]赵建新.传统豆酱发酵过程分析与控制发酵的研究[D].无锡:江南大学,2011.
[11]康旭,曾灿伟,黄红霞,等.酿造工艺对甜面酱风味成分影响研究[J].中国调味品,2011,36(1):58-60.
[12]孟鸳,乔宇,康旭,等.同时蒸馏萃取、溶剂萃取和顶空固相微萃取与GC-MS联用分析甜面酱的挥发性成分[J].中国调味品,2011,36(1):97-100.
[13]Kotiranta A,Lounatmaa K,Haapasalo M.Epidemiology and pathogenesis of Bacillus cereus infections[J].Microbes and Infection,2000(2):189-198.
[14]Abriouwl H,Maqueda M,Galvez A.Inhibition of bacterial growth,enterotoxin production and sporeout growth in strains of Bacillus cereus by bacteriocin AS-48[J].Appl Environ Mierobiol,2002,68(3):1473-1477.
[15]Zhou J S,Shu Q.Studies on translocation,acute oral toxicity and intestinal colonization of potentially probiotic lactic acid bacteria administ[J].Food Chem Toxieol,2000,38(2):153-161.
[16]吉志伟,刘阳,邓静,等.自然发酵甜面酱中一株芽孢杆菌的分离鉴定及益生特性初探[J].中国调味品,2015,40(12):26-30.
[17]Sekiguchi H,Watanabe M,Nakahara T,et al.Succession of bacterial community structure along the Changjiang River determined by denaturing gradient gel electrophoresis and clone library analysis[J].Appl Environ Microbiol,2002,68(10):5142-5150.