In Vitro Optimization of Enzymes Involved in Precorrin-2 Synthesis Using Response Surface Methodology
文献类型：期刊论文
| 作者 | Fang, Huan1,2,3; Dong, Huina1,2; Cai, Tao1; Zheng, Ping1,2; Li, Haixing4; Zhang, Dawei1,2; Sun, Jibin1,2 |
| 刊名 | PLOS ONE
 |
| 出版日期 | 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 |
| 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). |
入库方式： OAI收割
来源：天津工业生物技术研究所
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