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学科主题: Food Science & Technology
题名: Real-Time PCR Quantification of Protease-Producing Bacteria in Traditional Chinese Fish Sauce
作者: Xiao, YZ ; Zhao, SY ; Wu, DK ; Lin, WM ; Zhang, XY ; Gao, XY
通讯作者: gaoxiangyang@scau.edu.cn
关键词: Protease-producing bacteria ; 16S rDNA sequence ; Real-time PCR ; Fish sauce
刊名: FOOD ANALYTICAL METHODS
发表日期: 2014
卷: 7, 期:8, 页:1634-1642
收录类别: sci
部门归属: [Xiao, Yun-Zhu ; Zhao, Si-Yang ; Lin, Wei-Min ; Gao, Xiang-Yang] South China Agr Univ, Coll Food Sci, Guangzhou 510642, Guangdong, Peoples R China ; [Wu, Duan-Kai] Guangdong PRB Biotech Co Ltd, Guangzhou 510100, Guangdong, Peoples R China ; [Zhang, Xiao-Yong] Chinese Acad Sci, Key Lab Trop Marine Microbiol & Ecol, Guangdong Key Lab Marine Mat Med, RNAM Ctr Marine Microbiol,South China Sea Inst Oc, Guangzhou 510301, Guangdong, Peoples R China
项目归属: LMB
资助者: Shantou Fish Sauce Factory Co., Ltd.
摘要: This study aims to isolate, identify, and quantify protease-producing bacteria in traditional Chinese fish sauce. Fifty-five protease-producing bacteria were isolated from 10 fermented fish sauce samples in five growth media based on their morphology and caseinolytic activity. These isolates were identified through their 16S rDNA gene sequences. BLAST analysis of 16S rDNA sequences revealed that 46 and 9 strains belonged to Bacillus sp. and Virgibacillus halodenitrificans, respectively. We used EvaGreen dye in real-time PCR assay to target the partial bacterial 16S rDNA gene sequences of the 55 strains from fish sauce. Two primer pairs (A and B) were designed. Primer pair B was more suitable than primer pair A for real-time PCR assay, in which the optimum annealing temperature was 60 A degrees C. The significance of the developed method was proven by the highly linear characteristic of the standard curve that relates lower threshold cycle (Ct) values and 16S rDNA copy numbers of the standard DNA sample. This method was used to calculate the number of protease-producing bacteria in fish sauce. The minimum level of detection (1.44 x 10(3) copies/mu L) was also verified. The concentration of protease-producing bacteria in fish sauce was estimated to be (1.47 A +/- 0.73) x 10(3) colony-forming units (cfu)/mL by real-time PCR assay and showed a percentage of positive results correctly assigned of 91.8 % when compared to the plate culture method used as reference. The results may serve as a foundation for future studies on the microbial succession and variation of microorganisms during fish sauce fermentation.
原文出处: 查看原文
WOS记录号: WOS:000340050300010
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内容类型: 期刊论文
URI标识: http://ir.scsio.ac.cn/handle/344004/10394
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Xiao, YZ; Zhao, SY; Wu, DK; Lin, WM; Zhang, XY; Gao, XY.Real-Time PCR Quantification of Protease-Producing Bacteria in Traditional Chinese Fish Sauce,FOOD ANALYTICAL METHODS,2014,7(8):1634-1642
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