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Real-Time PCR Quantification of Protease-Producing Bacteria in Traditional Chinese Fish Sauce
Xiao, YZ; Zhao, SY; Wu, DK; Lin, WM; Zhang, XY; Gao, XY; gaoxiangyang@scau.edu.cn
2014
Source PublicationFOOD ANALYTICAL METHODS
ISSN1936-9751
Volume7Issue:8Pages:1634-1642
AbstractThis 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.
Department[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
KeywordProtease-producing Bacteria 16s Rdna Sequence Real-time Pcr Fish Sauce
Subject AreaFood Science & Technology
Funding OrganizationShantou Fish Sauce Factory Co., Ltd. ; Shantou Fish Sauce Factory Co., Ltd. ; Shantou Fish Sauce Factory Co., Ltd. ; Shantou Fish Sauce Factory Co., Ltd.
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Indexed Bysci
Funding ProjectLMB
Funding OrganizationShantou Fish Sauce Factory Co., Ltd. ; Shantou Fish Sauce Factory Co., Ltd. ; Shantou Fish Sauce Factory Co., Ltd. ; Shantou Fish Sauce Factory Co., Ltd.
WOS IDWOS:000340050300010
Citation statistics
Cited Times:6[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.scsio.ac.cn/handle/344004/10394
Collection中科院海洋生物资源可持续利用重点实验室
Corresponding Authorgaoxiangyang@scau.edu.cn
Recommended Citation
GB/T 7714
Xiao, YZ,Zhao, SY,Wu, DK,et al. Real-Time PCR Quantification of Protease-Producing Bacteria in Traditional Chinese Fish Sauce[J]. FOOD ANALYTICAL METHODS,2014,7(8):1634-1642.
APA Xiao, YZ.,Zhao, SY.,Wu, DK.,Lin, WM.,Zhang, XY.,...&gaoxiangyang@scau.edu.cn.(2014).Real-Time PCR Quantification of Protease-Producing Bacteria in Traditional Chinese Fish Sauce.FOOD ANALYTICAL METHODS,7(8),1634-1642.
MLA Xiao, YZ,et al."Real-Time PCR Quantification of Protease-Producing Bacteria in Traditional Chinese Fish Sauce".FOOD ANALYTICAL METHODS 7.8(2014):1634-1642.
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