Study and reengineering of the binding sites and allosteric regulation of biosynthetic threonine deaminase by isoleucine and valine in Escherichia coli
文献类型：期刊论文
| 作者 | Chen, Lin1,2,3; Chen, Zhen2,3,4; Zheng, Ping2,3,5,6; Sun, Jibin2,3,5,6; Zeng, An-Ping2,3,4 |
| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
 |
| 出版日期 | 2013-04-01 |
| 卷号 | 97期号:7页码:2939-2949 |
| 关键词 | Threonine deaminase Feedback inhibition L-isoleucine L-valine |
| 英文摘要 | Biosynthetic threonine deaminase (TD) is a key enzyme for the synthesis of isoleucine which is allosterically inhibited and activated by Ile and Val, respectively. The binding sites of Ile and Val and the mechanism of their regulations in TD are not clear, but essential for a rational design of efficient productive strain(s) for Ile and related amino acids. In this study, structure-based computational approach and site-directed mutagenesis were combined to identify the potential binding sites of Ile and Val in Escherichia coli TD. Our results demonstrated that each regulatory domain of the TD monomer possesses two nonequivalent effector-binding sites. The residues R362, E442, G445, A446, Y369, I460, and S461 only interact with Ile while E347, G350, and F352 are involved not only in the Ile binding but also in the Val binding. By further considering enzyme kinetic data, we propose a concentration-dependent mechanism of the allosteric regulation of TD by Ile and Val. For the construction of Ile overproducing strain, a novel TD mutant with double mutation of F352A/R362F was also created, which showed both higher activity and much stronger resistance to Ile inhibition comparing to those of wild-type enzyme. Overexpression of this mutant TD in E. coli JW3591 significantly increased the production of ketobutyrate and Ile in comparison to the reference strains overexpressing wild-type TD or the catabolic threonine deaminase (TdcB). This work builds a solid basis for the reengineering of TD and related microorganisms for Ile production. |
| 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] | CORYNEBACTERIUM-GLUTAMICUM ; ASPARTATE KINASE ; INHIBITION ; DEHYDRATASE ; EXPRESSION ; ENZYME ; ASPARTOKINASE ; PURIFICATION ; DEREGULATION ; MECHANISMS |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000316481800016 |
| 公开日期 | 2012-08-28 |
| 源URL | [http://localhost/handle/0/256]  |
| 专题 | 天津工业生物技术研究所_微生物代谢工程 张学礼_期刊论文 |
| 作者单位 | 1.Tianjin Univ, Dept Biochem Engn, Sch Chem Engn & Technol, Tianjin 300072, Peoples R China 2.IBB TUHH, Joint Lab Syst Biotechnol, Tianjin 300308, Peoples R China 3.TIB CAS, Tianjin 300308, Peoples R China 4.Hamburg Univ Technol IBB TUHH, Inst Bioproc & Biosyst Engn, D-21073 Hamburg, Germany 5.Chinese Acad Sci, Key Lab Syst Microbial Biotechnol, Tianjin 300308, Peoples R China 6.Chinese Acad Sci TIB CAS, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China |
| 推荐引用方式 GB/T 7714 | Chen, Lin,Chen, Zhen,Zheng, Ping,et al. Study and reengineering of the binding sites and allosteric regulation of biosynthetic threonine deaminase by isoleucine and valine in Escherichia coli[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2013,97(7):2939-2949. |
| APA | Chen, Lin,Chen, Zhen,Zheng, Ping,Sun, Jibin,&Zeng, An-Ping.(2013).Study and reengineering of the binding sites and allosteric regulation of biosynthetic threonine deaminase by isoleucine and valine in Escherichia coli.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,97(7),2939-2949. |
| MLA | Chen, Lin,et al."Study and reengineering of the binding sites and allosteric regulation of biosynthetic threonine deaminase by isoleucine and valine in Escherichia coli".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 97.7(2013):2939-2949. |
入库方式： OAI收割
来源：天津工业生物技术研究所
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