An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains

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	An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains
	Dong, Huina 1,2; Liu, Yongfei 1,2; Zu, Xin 1,4; Li, Ning 1; Li, Feiran 1; Zhang, Dawei 1,2,3
刊名	PLOS ONE
	2015-03-27
卷号	10 期号:3
英文摘要	Cytidine is an industrially useful precursor for the production of antiviral compounds and a variety of industrial compounds. Interest in the microbial production of cytidine has grown recently and high-throughput screening of cytidine over-producers is an important approach in large-scale industrial production using microorganisms. An enzymatic assay for cytidine was developed combining cytidine deaminase (CDA) and indophenol method. CDA catalyzes the cleavage of cytidine to uridine and NH3, the latter of which can be accurately determined using the indophenol method. The assay was performed in 96-well plates and had a linear detection range of cytidine of 0.058 - 10 mM. This assay was used to determine the amount of cytidine in fermentation flasks and the results were compared with that of High Perfomance Liquid Chromatography (HPLC) method. The detection range of the CDA method is not as wide as that of the HPLC, furthermore the correlation factor of CDA method is not as high as that of HPLC. However, it was suitable for the detection of large numbers of crude samples and was applied to high-throughput screening for high cytidine-producing strains using 96-well deep-hole culture plates. This assay was proved to be simple, accurate, specific and suitable for cytidine detection and high-throughput screening of cytidine-producing strains in large numbers of samples (96 well or more).
WOS标题词	Science & Technology
类目[WOS]	Multidisciplinary Sciences
研究领域[WOS]	Science & Technology - Other Topics
关键词[WOS]	BACILLUS-SUBTILIS ; GLUTAMATE-DEHYDROGENASE ; AMMONIA ; INDOPHENOL ; BERTHELOT ; DEAMINASE ; SYNTHETASE ; MUTANTS ; SERUM
收录类别	SCI
语种	英语
WOS记录号	WOS:000352133600109
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1448]
专题	天津工业生物技术研究所_微生物生理与蛋白质组学张大伟_期刊论文
作者单位	1.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China 2.Chinese Acad Sci, Key Lab Syst Microbial Biotechnol, Tianjin 300308, Peoples R China 3.Natl Engn Lab Ind Enzymes, Tianjin 300308, Peoples R China 4.Dalian Polytech Univ, Sch Biol Engn, Light Ind Technol & Engn, Dalian 116034, Peoples R China
推荐引用方式 GB/T 7714	Dong, Huina,Liu, Yongfei,Zu, Xin,et al. An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains[J]. PLOS ONE,2015,10(3).
APA	Dong, Huina,Liu, Yongfei,Zu, Xin,Li, Ning,Li, Feiran,&Zhang, Dawei.(2015).An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains.PLOS ONE,10(3).
MLA	Dong, Huina,et al."An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains".PLOS ONE 10.3(2015).