A fast and sensitive coupled enzyme assay for the measurement of L-threonine and application to high-throughput screening of threonine-overproducing strains

CORC > 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

	A fast and sensitive coupled enzyme assay for the measurement of L-threonine and application to high-throughput screening of threonine-overproducing strains
	Liu, Yongfei 1,2; Li, Feiran 1,2,4; Zhang, Xiaoran 1; Cao, Guoqiang 1; Jiang, Wenxia 1; Sun, Yuxiu 4; Zheng, Ping 1,2; Zhang, Dawei 1,2,3
刊名	ENZYME AND MICROBIAL TECHNOLOGY
	2014-12-01
卷号	67 期号:无页码:1-7
关键词	L-Threonine aldolase Threonine determination Coupled enzyme assay High throughput screening
英文摘要	Amino acids are important fermentation products that are used in the food, animal feed, pharmaceutical, and cosmetics industries. Development of a better quantitation assay to measure amino acid levels will facilitate research and application in medical and industrial fields. Here we report the development of an assay that allows the rapid measurement of threonine concentration. L-threonine aldolase (LTA) catalyzes the cleavage of threonine to glycine and acetaldehyde. The resulting acetaldehyde is further converted into ethanol by alcohol dehydrogenase accompanying NADH reduction. Therefore, monitoring the changes in NADH concentration can serve as a readout for the amount of threonine present within certain range. This LTA assay can distinguish a 0.5 mM difference in threonine concentration in some certain mediums. We successfully utilized this assay to measure L-threonine levels in industrial fermentation samples. Moreover, we were able to scale up this assay to 96-well format and use it in high-throughput screening of threonine-overproducing strains. We show this assay is simple, accurate, specific and suitable for determination of threonine concentration in large numbers of samples. (C) 2014 Elsevier Inc. All rights reserved.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biotechnology & Applied Microbiology
研究领域[WOS]	Biotechnology & Applied Microbiology
关键词[WOS]	ESCHERICHIA-COLI ; CORYNEBACTERIUM-GLUTAMICUM ; SERINE HYDROXYMETHYLTRANSFERASE ; SACCHAROMYCES-CEREVISIAE ; NUTRITIONAL QUALITY ; GLY1 GENE ; MUTANT ; ALDOLASES ; DEHYDROGENASE ; OPTIMIZATION
收录类别	SCI
语种	英语
WOS记录号	WOS:000345475800001
公开日期	2014-11-25
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/311007/494]
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.Tianjin Normal Univ, Coll Chem, Tianjin Key Lab Struct & Performance Funct Mol, Tianjin 300387, Peoples R China
推荐引用方式 GB/T 7714	Liu, Yongfei,Li, Feiran,Zhang, Xiaoran,et al. A fast and sensitive coupled enzyme assay for the measurement of L-threonine and application to high-throughput screening of threonine-overproducing strains[J]. ENZYME AND MICROBIAL TECHNOLOGY,2014,67(无):1-7.
APA	Liu, Yongfei.,Li, Feiran.,Zhang, Xiaoran.,Cao, Guoqiang.,Jiang, Wenxia.,...&Zhang, Dawei.(2014).A fast and sensitive coupled enzyme assay for the measurement of L-threonine and application to high-throughput screening of threonine-overproducing strains.ENZYME AND MICROBIAL TECHNOLOGY,67(无),1-7.
MLA	Liu, Yongfei,et al."A fast and sensitive coupled enzyme assay for the measurement of L-threonine and application to high-throughput screening of threonine-overproducing strains".ENZYME AND MICROBIAL TECHNOLOGY 67.无(2014):1-7.