Suh Wonchul
DuPont Central Research and Development, Wilmington, DE, USA.
Methods Mol Biol. 2012;834:49-62. doi: 10.1007/978-1-61779-483-4_4.
A noncarotenogenic microbe E. coli was engineered for high production of carotenoids. To increase the isoprenoid flux, the chromosomal native promoters of the rate-controlling steps (dxs, idi and ispDispF) in the isoprenoid pathway were replaced with a strong bacteriophage T5 promoter (P(T5)) by using the λ-Red recombinase system in combination with the Flp/FRT site-specific recombination system for marker excision and P1 transduction for gene trait stacking. The resulting high isoprenoid flux E. coli can be used as a starting strain to produce various carotenoids by introducing heterologous carotenoid genes. In this study, the high isoprenoid flux E. coli was transformed with a plasmid carrying the β-carotene biosynthetic genes from Pantoea stewartii for β-carotene production.
一种非类胡萝卜素生成微生物大肠杆菌被改造用于高效生产类胡萝卜素。为了增加类异戊二烯通量,通过使用λ-Red重组酶系统结合Flp/FRT位点特异性重组系统进行标记切除以及P1转导进行基因性状叠加,将类异戊二烯途径中速率控制步骤(dxs、idi和ispDispF)的染色体天然启动子替换为强噬菌体T5启动子(P(T5))。所得的高类异戊二烯通量大肠杆菌可作为起始菌株,通过引入异源类胡萝卜素基因来生产各种类胡萝卜素。在本研究中,用携带来自斯氏泛菌的β-胡萝卜素生物合成基因的质粒转化高类异戊二烯通量大肠杆菌以生产β-胡萝卜素。