蒲城椽头馍种子面团中酵母菌和乳酸菌的多样性分析
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摘要
为了研究蒲城椽头馍种子面团中的主要菌种酵母菌和乳酸菌的多样性,随机采取椽头馍传承人、西关椽头馍生产作坊、蒲城椽头馍厂、高阳村、九堂、石龙村、兴蒲清真食品厂及2个体户9份椽头馍种子面团进行分离、纯化、鉴定,得出酵母菌检出量在7.05×102~1.5×106CFU/g (干基),乳酸菌为1.08×103~1.94×106 CFU/g (干基)。通过26S r DNA D1/D2区和ITS区分析,54株酵母菌隶属于酿酒酵母,异常威克汉姆酵母,库德里阿兹威(氏)毕赤酵母,锁掷孢酵母。其中酿酒酵母在除高阳村外的8份样品中均有检出,且在系统发育树上聚集在一个由42株菌构成的大分支上,即优势酵母菌。7株乳酸菌通过16S rRNA序列分析,隶属于瑞士乳杆菌,戊糖片球菌,在西关椽头馍生产作坊和个体户2鉴定出5株瑞士乳杆菌,占71%,为优势菌。结果表明:蒲城椽头馍种子面团中酵母菌、乳酸菌具有一定的多样性,酿酒酵母和瑞士乳杆菌为其中的优势菌。
Chuantou steamed bread sourdough were sampled from Chuantou steamed bread heritage, Chuantou steamed bread production workshop in xiguan, Chuantou steamed bread factory, Gaoyang village, Nine hall, Shilong village, Xing Pu halal food factory and 2 self-employed and investigated the diversity of yeast and lactic acid bacteria by plate separation, purification and identification. The contents of yeast in the sourdoughs were from 4.03 ×102 to 1.8×106 CFU/g(dry base), while the lactic acid bacteria were from 1.08×103 to 1.94×106 CFU/g(dry base). Based on the results of 26 S rDNA D1/D2 region and Internal Transcribed Spacer(ITS) sequence amplification and sequencing, 54 yeast isolates were identified as Saccharomyces cerevisiae, Wickerhamomyces anomalus, Pichia kudriavzevii and Sporidiobolus pararoseus, respectively. S. cerevisiae in the 8 samples except Gaoyang village were detected, and the phylogenetic tree was gathered in a composed of 42 strains of branches, which was the advantage of yeast. Seven lactic acid bacteria(LAB) isolates were identified as Pediococcus pentosaceus(n=2) and Lactobacillus helveticus(n=5) in Chuantou steamed bread production workshop in xiguan and 2 self-employed according to their 16 S rRNA sequence. The results indicated that Saccharomyces and Lactobacillus were diverse in Chuantou steamed bread sourdoughs, among which, Saccharomyces cerevisiae and Lactobacillus helveticus were the dominant species.
Chuantou steamed bread sourdough were sampled from Chuantou steamed bread heritage, Chuantou steamed bread production workshop in xiguan, Chuantou steamed bread factory, Gaoyang village, Nine hall, Shilong village, Xing Pu halal food factory and 2 self-employed and investigated the diversity of yeast and lactic acid bacteria by plate separation, purification and identification. The contents of yeast in the sourdoughs were from 4.03 ×102 to 1.8×106 CFU/g(dry base), while the lactic acid bacteria were from 1.08×103 to 1.94×106 CFU/g(dry base). Based on the results of 26 S rDNA D1/D2 region and Internal Transcribed Spacer(ITS) sequence amplification and sequencing, 54 yeast isolates were identified as Saccharomyces cerevisiae, Wickerhamomyces anomalus, Pichia kudriavzevii and Sporidiobolus pararoseus, respectively. S. cerevisiae in the 8 samples except Gaoyang village were detected, and the phylogenetic tree was gathered in a composed of 42 strains of branches, which was the advantage of yeast. Seven lactic acid bacteria(LAB) isolates were identified as Pediococcus pentosaceus(n=2) and Lactobacillus helveticus(n=5) in Chuantou steamed bread production workshop in xiguan and 2 self-employed according to their 16 S rRNA sequence. The results indicated that Saccharomyces and Lactobacillus were diverse in Chuantou steamed bread sourdoughs, among which, Saccharomyces cerevisiae and Lactobacillus helveticus were the dominant species.
引文
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[3] ROBERT H, GABRIEL V, FONTAGN魪-FAUCHER C. Biodiversity of lactic acid bacteria in French wheat sourdough as determined by molecular characterization using species-specific PCR[J]. International Journal of Food Microbiology, 2009, 135(1):53-59.
[4]刘同杰,李云,吴诗榕,等.传统酸面团中细菌与酵母菌的分离与鉴定[J].现代食品科技, 2014, 26(9):114-120.
[5]米尔班古丽·阿卜杜如苏力,玛依古丽·库尔班,努斯热提古丽·安外尔,等.新疆传统馕发酵面团中酵母菌的多样性分析[J].食品科学, 2015, 36(19):199-203.
[6]郭刚,霍文捷,刘强,等.玉米秸秆青贮饲料发酵早期优良乳酸菌的分离和鉴定[J].畜牧与兽医,2016, 48(7):60-64.
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[9] ZHANG J C, LIU W J, SUN Z H, et al. Diversity of lactic acid bacteria and yeasts in traditional sourdoughs collected from western region in Inner Mongolia of China[J]. Food Control, 2011, 22(5):767-774.
[10]卿蔓君,白梅,张勇,等.西藏曲拉和云南乳饼中酵母菌的鉴定及其生物多样性[J].微生物学报,2010, 50(9):1141-1146.
[11] FELL J W, BOEKHOUT T, FONSECA A, et al.Biodiversity and systematics of basidiomycetous yeasts as determined by large-subunit rDNA D1/D2domain sequence analysis[J]. International Journal of Systematic and Evolutionary Microbiology, 2000, 50(3):1351-1371.
[12]潘向辉.酸面团中酵母菌筛选及发酵特性研究[D].保定:河北农业大学, 2011.
[13]明红梅,周健,陈蒙恩,等.大曲中异常威克汉姆酵母发酵产苯乙醇的条件优化[J].湖北农业科学,2015, 54(14):3492-3496.
[14] CODA R, CASSONE A, RIZZELLO C G, et al.Antifungal activity of Wickerhamomyces anomalus and Lactobacillus plantarum during sourdough fermentation:Identification of novel compounds and long-term effect during storage of wheat bread[J].Applied and Environmental Microbiology, 2011, 77(10):3484-3492.
[15] CHAN G F, GAN H M, LING H L, et al.Genome sequence of Pichia kudriavzevii M12, a potential producer of bioethanol and phytase[J]. Eukaryotic Cell, 2012, 11(10):1300-1301.
[16] GREPPI A, KRYCH L, COSTANTINI A, et al.Phytase-producing capacity of yeasts isolated from traditional African fermented food products and PHYPk gene expression of Pichia kudriavzevii strains[J]. International Journal of Food Microbiology,2015, 16(205):81-89.
[17]豆康宁,王飞,罗海澜.锁掷孢酵母菌粉挂面的研制[J].食品研究与开发, 2014, 35(20):53-56.
[18]莫海燕.饲用粪肠球菌、戊糖片球菌发酵工艺研究[D].石河子:石河子大学, 2013.
[19]句荣辉,黄广学,罗红霞.植物乳杆菌、戊糖片球菌对产肠毒素金黄色葡萄球菌生长的影响[J].肉类研究, 2013, 27(7):6-9.
[20]张丹丹,郭宇星,周慧敏,等.瑞士乳杆菌的益生特性[J].食品与发酵工业, 2014, 40(5):32-36.
[21]马先红,刘景圣,张文露,等.乳酸菌对中国传统主食馒头质量的影响[J].中国酿造, 2015, 34(11):10-13.
[2] LUANGSAKUL N, KEERATIPIBUL S, JINDAMORAKOT S, et al. Lactic acid bacteria and yeasts isolated from the starter doughs for Chinese steamed buns in Thailand[J]. LWT-Food Science and Technology, 2009, 42(8):1404-1412.
[3] ROBERT H, GABRIEL V, FONTAGN魪-FAUCHER C. Biodiversity of lactic acid bacteria in French wheat sourdough as determined by molecular characterization using species-specific PCR[J]. International Journal of Food Microbiology, 2009, 135(1):53-59.
[4]刘同杰,李云,吴诗榕,等.传统酸面团中细菌与酵母菌的分离与鉴定[J].现代食品科技, 2014, 26(9):114-120.
[5]米尔班古丽·阿卜杜如苏力,玛依古丽·库尔班,努斯热提古丽·安外尔,等.新疆传统馕发酵面团中酵母菌的多样性分析[J].食品科学, 2015, 36(19):199-203.
[6]郭刚,霍文捷,刘强,等.玉米秸秆青贮饲料发酵早期优良乳酸菌的分离和鉴定[J].畜牧与兽医,2016, 48(7):60-64.
[7] SCHEIRLINCK I, MEULEN R V D, SCHOOR A V, et al. Influence of geographical origin and flour type on diversity of lactic acid bacteria in traditional Belgian sourdoughs[J]. Applied and Environmental Microbiology, 2007, 73(19):6262-6269.
[8]李峰,贾建波.分析生物学实验[M].武汉:华中科技大学出版社, 2014:125-129.
[9] ZHANG J C, LIU W J, SUN Z H, et al. Diversity of lactic acid bacteria and yeasts in traditional sourdoughs collected from western region in Inner Mongolia of China[J]. Food Control, 2011, 22(5):767-774.
[10]卿蔓君,白梅,张勇,等.西藏曲拉和云南乳饼中酵母菌的鉴定及其生物多样性[J].微生物学报,2010, 50(9):1141-1146.
[11] FELL J W, BOEKHOUT T, FONSECA A, et al.Biodiversity and systematics of basidiomycetous yeasts as determined by large-subunit rDNA D1/D2domain sequence analysis[J]. International Journal of Systematic and Evolutionary Microbiology, 2000, 50(3):1351-1371.
[12]潘向辉.酸面团中酵母菌筛选及发酵特性研究[D].保定:河北农业大学, 2011.
[13]明红梅,周健,陈蒙恩,等.大曲中异常威克汉姆酵母发酵产苯乙醇的条件优化[J].湖北农业科学,2015, 54(14):3492-3496.
[14] CODA R, CASSONE A, RIZZELLO C G, et al.Antifungal activity of Wickerhamomyces anomalus and Lactobacillus plantarum during sourdough fermentation:Identification of novel compounds and long-term effect during storage of wheat bread[J].Applied and Environmental Microbiology, 2011, 77(10):3484-3492.
[15] CHAN G F, GAN H M, LING H L, et al.Genome sequence of Pichia kudriavzevii M12, a potential producer of bioethanol and phytase[J]. Eukaryotic Cell, 2012, 11(10):1300-1301.
[16] GREPPI A, KRYCH L, COSTANTINI A, et al.Phytase-producing capacity of yeasts isolated from traditional African fermented food products and PHYPk gene expression of Pichia kudriavzevii strains[J]. International Journal of Food Microbiology,2015, 16(205):81-89.
[17]豆康宁,王飞,罗海澜.锁掷孢酵母菌粉挂面的研制[J].食品研究与开发, 2014, 35(20):53-56.
[18]莫海燕.饲用粪肠球菌、戊糖片球菌发酵工艺研究[D].石河子:石河子大学, 2013.
[19]句荣辉,黄广学,罗红霞.植物乳杆菌、戊糖片球菌对产肠毒素金黄色葡萄球菌生长的影响[J].肉类研究, 2013, 27(7):6-9.
[20]张丹丹,郭宇星,周慧敏,等.瑞士乳杆菌的益生特性[J].食品与发酵工业, 2014, 40(5):32-36.
[21]马先红,刘景圣,张文露,等.乳酸菌对中国传统主食馒头质量的影响[J].中国酿造, 2015, 34(11):10-13.