胀桶番茄酱中腐败微生物的分离及鉴定
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摘要
为了研究胀桶番茄酱中腐败微生物种类,采用梯度稀释法、选择性培养法和划线纯化法对番茄酱中主要的腐败微生物进行分离纯化,利用形态学特征观察、16S rDNA和ITS序列分析,并结合生理生化性质对分离菌株进行鉴定。结果显示编号为B001、B002、B003、B004的4株细菌有1株厚壁类芽孢杆菌(Paenibacillus cineris)、2株蜡样芽胞杆菌(Bacillus cereus)和1株蕈状芽孢杆菌(Bacillus mycoides)。标号为F001的1株酵母为奥默毕赤酵母(Pichia kudriavzevil)。因此,胀桶番茄酱中的主要腐败微生物为厚壁类芽孢杆菌、蜡样芽胞杆菌、蕈状芽孢杆菌和奥默毕赤酵母。
In order to obtain the spoilage microorganisms in the swell barrelled tomato sauce,the main spoilage microorganisms in tomato sauce were separated and purified by gradient dilution method,selective culture method and scribing purification method. The morphological characteristics and 16 S rDNA and ITS sequence analysis were used. The isolates were identified by combining physiological and biochemical identification as well. The results showed that the four bacteria numbered B001,B002,B003 and B004 included one Paenibacillus cineris,two Bacillus cereus,and one Bacillus mycoides. One yeast strain named F001 was Pichia kudriavzevil. Therefore,the main spoilage microorganisms in the swell barrelled tomato sauce should be Bacillus licheniformis,Bacillus cereus,Bacillus licheniformis and Pichia kudriavzevil.
In order to obtain the spoilage microorganisms in the swell barrelled tomato sauce,the main spoilage microorganisms in tomato sauce were separated and purified by gradient dilution method,selective culture method and scribing purification method. The morphological characteristics and 16 S rDNA and ITS sequence analysis were used. The isolates were identified by combining physiological and biochemical identification as well. The results showed that the four bacteria numbered B001,B002,B003 and B004 included one Paenibacillus cineris,two Bacillus cereus,and one Bacillus mycoides. One yeast strain named F001 was Pichia kudriavzevil. Therefore,the main spoilage microorganisms in the swell barrelled tomato sauce should be Bacillus licheniformis,Bacillus cereus,Bacillus licheniformis and Pichia kudriavzevil.
引文
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[15]姚晓瑞宁,高飞飞,王斌,等. 库尔勒香梨表皮酵母菌的筛选及耐受性分析[J].中国酿造,2017,36(6):67-71.
[16]陆文俊,王芳,陆兆新,等. 食品中酵母菌的分离鉴定及其致腐能力[J].食品科学,2016,37(19):177-182.
[17]闫国宏,傅力,古丽娜孜. 疑似胀袋番茄酱中几种微生物的检测研究[J].食品科技,2007(12):187-189.
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[2]朱宁,马骥. 中国番茄酱国际竞争力比较分析与提升路径[J]. 世界农业,2014:138-142.
[3]杨红红,陈国刚,刘娅,等. 桶装胀罐番茄酱中腐败菌分离纯化及鉴定研究[J].食品工业科技,2014,25(1):164-173.
[4]黄玲,黄忠梅,吴海文. 胀袋番茄酱中分离出季也蒙假丝酵母和丛生丝孢酵母[J].中国食品卫生杂志,2006,18(6):547-548.
[5]Vercammen A,Vivijs B,Lurquin I,et al. Germination and inactivation of Bacillus coagulans and Alicyclobacillus acidoterrestris spores by high hydrostatic pressure treatment in buffer and tomato sauce[J].International Journal of Food Microbiology,2012,152(3):162-167.
[6]李慧,陈历水,刘洋,等. 坛装黄酒贮存过程中污染微生物的分离鉴定[J].中国酿造,2018,5:254-258.
[7]Chin H S,Fred B,Fleming H P,et al. Identifications of predominant bacterial isolates from the fermenting Kinchi using ITS-PCR and partial 16S rDNA sequence analyses[J]. Microbiol Biotechnol,2001,16(1):68-76.
[8]任清,候昌. 北宗黄酒麦曲微生物的分离鉴定[J].食品科学,2017,38(4):77-82.
[9]李慧,蔡军,胡梦龙,等. 产黄曲霉毒素真菌多重PCR检测体系的建立[J].2017,32(1):125-129.
[10]Chen L X,Yang Y W,Zhou H,et al. Isolation and identification of microorganism affecting eating quality in fermented fresh rice noodle[J]. Food & Machinery,2018,34(1):33-43.
[11]吴海文,黄玲,傅力.番茄酱中好氧细菌分离方法的研究[J]. 新疆农业科学,2006,43(1):63-68.
[12]李靖,马嫄,徐丹,等. 红油豆瓣酱中产气芽孢杆菌的分离、鉴定与抑制研究[J]. 中国酿造,2017,36(8):31-35.
[13]Fell J W,Boekhout T,Fonseca A,et al. Biodiversity and systematics of basidiomycetous yeasts as determined by large-subunit rDNA D1/D2 domain sequence analysis[J]. International Journal of Systematic & Evolutionary Microbiology,2000,50(3):1351-1371.
[14]龙雪峰,陈卫军,王挥,等. 低温贮藏椰肉品质变化及腐败菌分离与鉴定[J].广东农业科学,2016(9):108-112.
[15]姚晓瑞宁,高飞飞,王斌,等. 库尔勒香梨表皮酵母菌的筛选及耐受性分析[J].中国酿造,2017,36(6):67-71.
[16]陆文俊,王芳,陆兆新,等. 食品中酵母菌的分离鉴定及其致腐能力[J].食品科学,2016,37(19):177-182.
[17]闫国宏,傅力,古丽娜孜. 疑似胀袋番茄酱中几种微生物的检测研究[J].食品科技,2007(12):187-189.
[18]丁生林,徐榕蔓. 番茄酱产气变质成因分析及控制要点[J].食品与发酵工业,2010,136(9):90-92.