Gupta Shweta, Marko Melissa G, Miller Vandana A, Schaefer Frederick T, Anthony Jennifer R, Porter John R
Department of Biological Sciences, University of the Sciences, 600 South Forty-Third Street, Philadelphia, PA, 19104, USA,
Appl Biochem Biotechnol. 2015 Mar;175(5):2319-31. doi: 10.1007/s12010-014-1382-4. Epub 2014 Dec 9.
Metabolic engineering of heterologous pathways has allowed the production of therapeutically important compounds in microbial systems. Here, we report the engineering of a monoterpenoid biosynthetic pathway into Escherichia coli. Five genes encoding sequential enzymes for perillyl alcohol biosynthesis from the precursors isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) were engineered into E. coli. Expression of these genes allowed the production of the intermediate limonene, but the downstream monoterpenoid, perillyl alcohol, was not detected. A new compound was detected but could not be identified based on the data obtained. Only 1.6 μg/ml of the compound was being produced from the engineered E. coli strain, but, when these cultures were fed limonene as a substrate, the production was nearly 250 μg/ml. This unknown compound inhibited the cell proliferation of MCF-7 and MDA-MB-231 breast cancer cells in 48-h treatment experiments. This compound may have potential benefits in breast cancer treatment. This is the first report showing the production of a monoterpenoid in engineered E. coli and its antiproliferative effects in breast cancer cells.
异源途径的代谢工程已使微生物系统能够生产具有治疗重要性的化合物。在此，我们报告了将单萜生物合成途径工程化导入大肠杆菌的情况。五个编码从异戊烯基焦磷酸（IPP）和二甲基烯丙基焦磷酸（DMAPP）前体生物合成紫苏醇的顺序酶的基因被工程化导入大肠杆菌。这些基因的表达使得中间产物柠檬烯得以产生，但未检测到下游的单萜紫苏醇。检测到一种新化合物，但根据所获得的数据无法鉴定。工程化的大肠杆菌菌株仅产生1.6μg/ml的该化合物，但是，当用柠檬烯作为底物投喂这些培养物时，产量接近250μg/ml。在48小时的处理实验中，这种未知化合物抑制了MCF-7和MDA-MB-231乳腺癌细胞的增殖。这种化合物可能在乳腺癌治疗中具有潜在益处。这是首次报道在工程化大肠杆菌中生产单萜及其对乳腺癌细胞的抗增殖作用。
