Department of Chemical and Biological Engineering, Science and Technology Center, Tufts University, 4 Colby Street, Medford, MA 02155, USA.
J Ind Microbiol Biotechnol. 2011 Nov;38(11):1809-20. doi: 10.1007/s10295-011-0969-9. Epub 2011 Apr 13.
Natural products have long served as rich sources of drugs possessing a wide range of pharmacological activities. The discovery and development of natural product drug candidates is often hampered by the inability to efficiently scale and produce a molecule of interest, due to inherent qualities of the native producer. Heterologous biosynthesis in an engineering and process-friendly host emerged as an option to produce complex natural products. Escherichia coli has previously been utilized to produce complex precursors to two popular natural product drugs, erythromycin and paclitaxel. These two molecules represent two of the largest classes of natural products, polyketides and isoprenoids, respectively. In this study, we have developed a platform E. coli strain capable of simultaneous production of both product precursors at titers greater than 15 mg l(-1). The utilization of a two-phase batch bioreactor allowed for very strong in situ separation (having a partitioning coefficient of greater than 5,000), which would facilitate downstream purification processes. The system developed here could also be used in metagenomic studies to screen environmental DNA for natural product discovery and preliminary production experiments.
天然产物长期以来一直是具有广泛药理活性的药物的丰富来源。由于天然产物的固有特性，难以有效地规模化生产感兴趣的分子，这常常阻碍了天然产物候选药物的发现和开发。在一个工程友好型和易于处理的宿主中进行异源生物合成成为生产复杂天然产物的一种选择。大肠杆菌以前曾被用于生产两种广受欢迎的天然产物药物——红霉素和紫杉醇的复杂前体。这两种分子分别代表了最大的两类天然产物，聚酮类和异戊二烯类。在这项研究中，我们开发了一种大肠杆菌菌株平台，能够以大于 15 毫克/升的浓度同时生产这两种产物前体。两相分批生物反应器的使用允许非常强的原位分离（具有大于 5000 的分配系数），这将有利于下游的纯化过程。这里开发的系统也可用于宏基因组学研究，以筛选环境 DNA 进行天然产物的发现和初步生产实验。
