Improving the Production of L-Phenylalanine by Identifying Key Enzymes Through Multi-Enzyme Reaction System in Vitro

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	Improving the Production of L-Phenylalanine by Identifying Key Enzymes Through Multi-Enzyme Reaction System in Vitro
	Ding, Dongqin 1,2; Liu, Yongfei 1,3; Xu, Yiran 1; Zheng, Ping 1,3; Li, Haixing 2; Zhang, Dawei 1,3; Sun, Jibin 1,3
刊名	SCIENTIFIC REPORTS
	2016-08-25
卷号	6
英文摘要	L-Phenylalanine (L-Phe) is an important amino acid used in both food and medicinal applications. We developed an in vitro system that allowed a direct, quantitative investigation of phenylalanine biosynthesis in E. coli. Here, the absolute concentrations of six enzymes (AroK, AroL, AroA, AroC, PheA and TyrB) involved in the shikimate (SHIK) pathway were determined by a quantitative proteomics approach and in vitro enzyme titration experiments. The reconstitution of an in vitro reaction system for these six enzymes was established and their effects on the phenylalanine production were tested. The results showed that the yield of phenylalanine increased 3.0 and 2.1 times when the concentrations of shikimate kinase (AroL) and 5-enolpyruvoyl shikimate 3-phosphate (EPSP) synthase (AroA) were increased 2.5 times. Consistent results were obtained from in vivo via the overexpression of AroA in a phenylalanine-producing strain, and the titer of phenylalanine reached 62.47 g/l after 48 h cultivation in a 5-liter jar fermentor. Our quantitative findings provide a practical method to detect the potential bottleneck in a specific metabolic pathway to determine which gene products should be targeted to improve the yield of the desired product.
WOS标题词	Science & Technology
类目[WOS]	Multidisciplinary Sciences
研究领域[WOS]	Science & Technology - Other Topics
关键词[WOS]	ESCHERICHIA-COLI K-12 ; 5-ENOLPYRUVYLSHIKIMATE 3-PHOSPHATE SYNTHASE ; MUTASE-PREPHENATE DEHYDRATASE ; AMINO-ACID-SEQUENCE ; SHIKIMATE KINASE-II ; CHORISMATE MUTASE ; PURIFICATION ; GENE ; ASPARTATE ; AROL
收录类别	SCI
语种	英语
WOS记录号	WOS:000381856300001
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/2918]
专题	天津工业生物技术研究所_系统与合成生物技术实验室孙际宾_期刊论文
作者单位	1.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China 2.Nanchang Univ, Sch Food, Dept Food Sci & Engn, Nanchang 330029, Peoples R China 3.Chinese Acad Sci, Key Lab Syst Microbial Biotechnol, Tianjin 300308, Peoples R China
推荐引用方式 GB/T 7714	Ding, Dongqin,Liu, Yongfei,Xu, Yiran,et al. Improving the Production of L-Phenylalanine by Identifying Key Enzymes Through Multi-Enzyme Reaction System in Vitro[J]. SCIENTIFIC REPORTS,2016,6.
APA	Ding, Dongqin.,Liu, Yongfei.,Xu, Yiran.,Zheng, Ping.,Li, Haixing.,...&Sun, Jibin.(2016).Improving the Production of L-Phenylalanine by Identifying Key Enzymes Through Multi-Enzyme Reaction System in Vitro.SCIENTIFIC REPORTS,6.
MLA	Ding, Dongqin,et al."Improving the Production of L-Phenylalanine by Identifying Key Enzymes Through Multi-Enzyme Reaction System in Vitro".SCIENTIFIC REPORTS 6(2016).