Abstract

The uncoordinated expression of a target biosynthetic pathway often results in low growth and productivity for a producing strain due to the accumulation of toxic metabolic intermediates. In this study, we report an increase in isoprene accumulation by an engineered strain of Pantoea ananatis that possesses foreign, artificial mevalonate-pathway genes. This was achieved due to the simultaneous, φ80-Int-dependent integration of a mixture of conditionally replicated plasmids carrying genes for the upper and lower mevalonate pathways and mevalonate kinase, followed by selection according to the isoprene accumulation of clones with optimal copy numbers of the integrative cassettes. A previously developed method, Dual In/Out, was modified to accelerate the construction of recipient strains containing several attB sites of the φ80 phage in their genomes.

中文翻译：

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由于甲羟戊酸途径基因的平衡扩增，重组Pantoea ananatis菌株提高了异戊二烯的产量

摘要

由于有毒代谢中间体的积累，靶生物合成途径的不协调表达常常导致生产菌株的生长和生产率低下。在这项研究中，我们报告了具有外来的人工甲羟戊酸途径基因的泛酸工程菌的工程菌株增加了异戊二烯的积累。之所以能够做到这一点，是因为同时进行了条件复制质粒的混合物的φ80-Int依赖性整合，这些质粒带有上，下甲羟戊酸途径和甲羟戊酸激酶的基因，然后根据具有最佳拷贝数的克隆的异戊二烯进行选择集成盒式磁带。修改了以前开发的双重输入/输出方法，以加快包含多个受体菌株的构建φ80噬菌体的attB位点在其基因组中。