In Vitro Optimization of Enzymes Involved in Precorrin-2 Synthesis Using Response Surface Methodology

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	In Vitro Optimization of Enzymes Involved in Precorrin-2 Synthesis Using Response Surface Methodology
	Fang, Huan 1,2,3; Dong, Huina 1,2; Cai, Tao 1; Zheng, Ping 1,2; Li, Haixing 4; Zhang, Dawei 1,2; Sun, Jibin 1,2
刊名	PLOS ONE
	2016-03-14
卷号	11 期号:3
英文摘要	In order to maximize the production of biologically-derived chemicals, kinetic analyses are first necessary for predicting the role of enzyme components and coordinating enzymes in the same reaction system. Precorrin-2 is a key precursor of cobalamin and siroheme synthesis. In this study, we sought to optimize the concentrations of several molecules involved in precorrin-2 synthesis in vitro: porphobilinogen synthase (PBGS), porphobilinogen deaminase (PBGD), uroporphyrinogen III synthase (UROS), and S-adenosyl-l-methionine-dependent urogen III methyltransferase (SUMT). Response surface methodology was applied to develop a kinetic model designed to maximize precorrin-2 productivity. The optimal molar ratios of PBGS, PBGD, UROS, and SUMT were found to be approximately 1: 7: 7: 34, respectively. Maximum precorrin-2 production was achieved at 0.1966 +/- 0.0028 mu M/min, agreeing with the kinetic model's predicted value of 0.1950 mu M/min. The optimal concentrations of the cofactor S-adenosyl-L-methionine (SAM) and substrate 5-aminolevulinic acid (ALA) were also determined to be 200 mu M and 5 mM, respectively, in a tandem-enzyme assay. By optimizing the relative concentrations of these enzymes, we were able to minimize the effects of substrate inhibition and feedback inhibition by S-adenosylhomocysteine on SUMT and thereby increase the production of precorrin-2 by approximately five-fold. These results demonstrate the effectiveness of kinetic modeling via response surface methodology for maximizing the production of biologically-derived chemicals.
WOS标题词	Science & Technology
类目[WOS]	Multidisciplinary Sciences
研究领域[WOS]	Science & Technology - Other Topics
关键词[WOS]	UROPORPHYRINOGEN-III METHYLTRANSFERASE ; ADENOSYL-L-METHIONINE ; HEME D(1) BIOSYNTHESIS ; ESCHERICHIA-COLI ; BACILLUS-MEGATERIUM ; FATTY-ACID ; EXPRESSION ; PATHWAY ; GENE ; OVERPRODUCTION
收录类别	SCI
语种	英语
WOS记录号	WOS:000372570600052
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/2237]
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, Peoples R China 2.Chinese Acad Sci, Key Lab Syst Microbial Biotechnol, Tianjin, Peoples R China 3.Univ Chinese Acad Sci, Beijing, Peoples R China 4.Nanchang Univ, Sinogerman Joint Res Inst, Nanchang, Jiangxi, Peoples R China
推荐引用方式 GB/T 7714	Fang, Huan,Dong, Huina,Cai, Tao,et al. In Vitro Optimization of Enzymes Involved in Precorrin-2 Synthesis Using Response Surface Methodology[J]. PLOS ONE,2016,11(3).
APA	Fang, Huan.,Dong, Huina.,Cai, Tao.,Zheng, Ping.,Li, Haixing.,...&Sun, Jibin.(2016).In Vitro Optimization of Enzymes Involved in Precorrin-2 Synthesis Using Response Surface Methodology.PLOS ONE,11(3).
MLA	Fang, Huan,et al."In Vitro Optimization of Enzymes Involved in Precorrin-2 Synthesis Using Response Surface Methodology".PLOS ONE 11.3(2016).