Targeted cofactor quantification in metabolically engineered E-coli using solid phase extraction and hydrophilic interaction liquid chromatography-mass spectrometry

CORC > 天津工业生物技术研究所 > 生物质谱与定量生物学研究水雯箐 > 期刊论文

	Targeted cofactor quantification in metabolically engineered E-coli using solid phase extraction and hydrophilic interaction liquid chromatography-mass spectrometry
	Li, Zhucui 1; Yang, Afang 2; Li, Yujing 3; Liu, Pingping 1; Zhang, Zhidan 1; Zhang, Xueli 1; Shui, Wenqing 1
刊名	JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
	2016-03-01
卷号	1014 页码:107-115
关键词	SPE HILIC-MS Cofactor Targeted metabolome Energy metabolism Isobutanol production
英文摘要	Quantification of energy and redox cofactors is of great value to synthetic biologists to infer the balance of energy metabolism in engineered microbial strains and assess each strain's potential for further improvement. Most currently used methods for intracellular cofactor measurement suffer from incomplete coverage, low reproducibility, suboptimal sensitivity or specificity. In this study, we described an SPE-HILIC/MS approach for simultaneous determination of six cofactor targets (ATP, ADP, NAD, NADH, NADP, NADPH) in Escherichia coli cells. Sufficient linearity, precision and metabolite recoveries of this new approach justified its reliability in targeted cofactor quantification. Our approach was then compared with conventional enzymatic assays to demonstrate its superior performance. We applied the SPE-HILIC/MS approach to profile shift of cofactor balances in several engineered E. coli strains with varying isobutanol production. Our cofactor analysis clearly revealed that optimal energy fitness was achieved in the highest-yield strain through combined modulation of a transhydrogenase and a NAD(+) kinase. Apart from the targeted cofactors, the SPE enrichment procedure also allowed for confident identification of 39 groups of polar metabolites mainly involved in central carbon metabolism in E. coli cells. (C) 2016 Published by Elsevier B.V.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences
类目[WOS]	Biochemical Research Methods ; Chemistry, Analytical
研究领域[WOS]	Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]	TOF-MS ; NADH ; METABOLOMICS ; URINE ; DRUGS ; SEPARATION ; EVOLUTION ; SUCCINATE ; COVERAGE ; NAD(+)
收录类别	SCI
语种	英语
WOS记录号	WOS:000371549900015
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1525]
专题	天津工业生物技术研究所_生物质谱与定量生物学研究水雯箐_期刊论文
作者单位	1.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Key Lab Syst Microbial Biotechnol, Tianjin 300308, Peoples R China 2.Tianjin Univ Sci & Technol, Coll Bioengn, Tianjin 300457, Peoples R China 3.Nankai Univ, Coll Life Sci, Tianjin 300071, Peoples R China
推荐引用方式 GB/T 7714	Li, Zhucui,Yang, Afang,Li, Yujing,et al. Targeted cofactor quantification in metabolically engineered E-coli using solid phase extraction and hydrophilic interaction liquid chromatography-mass spectrometry[J]. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,2016,1014:107-115.
APA	Li, Zhucui.,Yang, Afang.,Li, Yujing.,Liu, Pingping.,Zhang, Zhidan.,...&Shui, Wenqing.(2016).Targeted cofactor quantification in metabolically engineered E-coli using solid phase extraction and hydrophilic interaction liquid chromatography-mass spectrometry.JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,1014,107-115.
MLA	Li, Zhucui,et al."Targeted cofactor quantification in metabolically engineered E-coli using solid phase extraction and hydrophilic interaction liquid chromatography-mass spectrometry".JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 1014(2016):107-115.