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利用工程大肠杆菌从葡萄糖高效生产 5-氨基乙酰丙酸。

Engineering Escherichia coli for efficient production of 5-aminolevulinic acid from glucose.

机构信息

State Key Laboratory of Microbial Technology, Shandong University, Jinan, People's Republic of China.

出版信息

Metab Eng. 2011 Sep;13(5):492-8. doi: 10.1016/j.ymben.2011.05.003. Epub 2011 May 23.

Abstract

5-Aminolevulinic acid (ALA) recently received much attention due to its potential applications in many fields. In this study, we developed a metabolic strategy to produce ALA directly from glucose in recombinant Escherichia coli via the C5 pathway. The expression of a mutated hemA gene, encoding a glutamyl-tRNA reductase from Salmonella arizona, significantly improved ALA production from 31.1 to 176mg/L. Glutamate-1-semialdehyde aminotransferase from E. coli was found to have a synergistic effect with HemA(M) from S. arizona on ALA production (2052mg/L). In addition, we identified a threonine/homoserine exporter in E. coli, encoded by rhtA gene, which exported ALA due to its broad substrate specificity. The constructed E. coli DALA produced 4.13g/L ALA in modified minimal medium from glucose without adding any other co-substrate or inhibitor. This strategy offered an attractive potential to metabolic production of ALA in E. coli.

摘要

5-氨基乙酰丙酸(ALA)由于其在许多领域的潜在应用而受到广泛关注。在这项研究中,我们通过 C5 途径开发了一种代谢策略,使重组大肠杆菌能够直接从葡萄糖生产 ALA。表达突变的 hemA 基因,编码来自亚利桑那沙门氏菌的谷氨酰-tRNA 还原酶,可将 ALA 的产量从 31.1mg/L 显著提高到 176mg/L。发现来自大肠杆菌的谷氨酸-1-半醛氨基转移酶与来自 S. arizona 的 HemA(M) 对 ALA 的产生具有协同作用(2052mg/L)。此外,我们鉴定了大肠杆菌中的一种苏氨酸/高丝氨酸外排蛋白,由 rhtA 基因编码,由于其广泛的底物特异性,它可将 ALA 排出细胞。构建的大肠杆菌 DALA 在改良的最小培养基中从葡萄糖生产 4.13g/L 的 ALA,无需添加任何其他共底物或抑制剂。该策略为大肠杆菌中 ALA 的代谢生产提供了有吸引力的潜力。

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