Improving biosynthetic production of pinene through plasmid recombination elimination and pathway optimization
Improving biosynthetic production of pinene through plasmid recombination elimination and pathway optimization
复制标题
通过质粒重组消除和途径优化提高蒎烯的生物合成产量。
DOI：
10.1016/j.plasmid.2019.102431
复制
发表时间：
2019-09-01
期刊：
影响因子：
2.6
通讯作者：
Meng, Er
中科院分区：
文献类型：
--
作者：
Bao, Shao-Heng;Zhang, Dong-Yi;Meng, Er
萜烯是一种工业用途广泛的单萜烯，特别是作为一种很有前途的高能量密度喷气燃料。传统的工业生产工艺耗材大，产率低。作为另一种选择，微生物已经通过先进的合成生物技术进行了工程改造，以生产各种异源产品，包括蒎烯。在这里，我们研究了在发酵过程中编码Pinene产生途径的遗传电路的稳定性及其与Pinene滴度的关系。通过替换遗传元件中的SCAR序列和修改大肠杆菌MG1655的基因组，消除了由于严重的代谢负担而导致的质粒丢失，从而显著提高了蒎烯效价。此外，通过酶的过度表达和中间体的监测，分析了异源甲戊酸途径。将优化后的途径质粒和菌株组合，可使Pinene效价提高到104.6 mg/L。
Pinene is a monoterpene with wide industrial applications, especially as a promising high energy-density jet fuel. Traditional production of pinene on an industrial scale is material consumptive and has a low yield. As an alternative, microbial organisms have been engineered though advanced synthetic biological techniques to produce a variety of heterologous products, including pinene. Here, we investigated the stability of genetic circuits encoding the pinene producing pathway during fermentation and its relationship to the pinene titer. By replacing scar sequences in the genetic elements and modifying the genome of E. coli strain MG1655, plasmid loss caused by serious metabolic burden was eliminated, generating a remarkable increase in the pinene titer. Furthermore, the heterologous mevalonate pathway was analyzed by overexpression of enzymes and intermediates monitoring. Optimized pathway plasmids and strains were combined to increase the pinene titer to 104.6 mg/L.