A Pressure Test to Make 10 Molecules in 90 Days: External Evaluation of Methods to Engineer Biology
A Pressure Test to Make 10 Molecules in 90 Days: External Evaluation of Methods to Engineer Biology
复制标题
DOI：
10.1021/jacs.7b13292
复制
发表时间：
2018-03-28
影响因子：
15
通讯作者：
Gordon, D. Benjamin
中科院分区：
文献类型：
--
作者：
Casini, Arturo;Chang, Fang-Yuan;Gordon, D. Benjamin
基因工程的集中设施或“生物铸造厂”提供了设计生物体以满足医学、农业、工业和国防新兴需求的潜力。该领域的技术进步很快，但很难衡量当前的能力或确定项目之间的差距。为此，我们的铸造厂通过定时“压力测试”进行了评估，其中有 3 个月的时间来构建生物体，以生产我们事先未知的 10 种分子。通过应用多种新方法，我们在执行期间为十分之六的目标生产出了所需的分子或密切相关的分子，并在其他分子的生产方面取得了进展。具体来说，我们提高了 1-十六醇、吡咯尼群和帕西霉素 D 的效价，发现了强效抗菌剂烯二炔弹头的新途径，建立了用于单萜生产的无细胞系统，生产了长春新碱生物合成的中间体，并编码了 DNA 库中 540 个双吲哚的 7802 条可单独检索的途径。设计并构建了四氢呋喃和巴巴酰胺的途径，但毒性或分析工具阻碍了进一步的进展。总之，我们构建了 1.2 Mb DNA，构建了跨越 5 个物种（酿酒酵母、大肠杆菌、白黄链霉菌、天蓝色链霉菌和白链霉菌）的 215 个菌株，建立了两个无细胞系统，并进行了 690 次内部开发的分子测定。
Centralized facilities for genetic engineering, or "biofoundries", offer the potential to design organisms to address emerging needs in medicine, agriculture, industry, and defense. The field has seen rapid advances in technology, but it is difficult to gauge current capabilities or identify gaps across projects. To this end, our foundry was assessed via a timed "pressure test", in which 3 months were given to build organisms to produce 10 molecules unknown to us in advance. By applying a diversity of new approaches, we produced the desired molecule or a closely related one for six out of 10 targets during the performance period and made advances toward production of the others as well. Specifically, we increased the titers of 1-hexadecanol, pyrrolnitrin, and pacidamycin D, found novel routes to the enediyne warhead underlying powerful antimicrobials, established a cell-free system for monoterpene production, produced an intermediate toward vincristine biosynthesis, and encoded 7802 individually retrievable pathways to 540 bisindoles in a DNA pool. Pathways to tetrahydrofuran and barbamide were designed and constructed, but toxicity or analytical tools inhibited further progress. In sum, we constructed 1.2 Mb DNA, built 215 strains spanning five species (Saccharomyces cerevisiae, Escherichia coli, Streptomyces albidoflavus, Streptomyces coelicolor, and Streptomyces albovinaceus), established two cell-free systems, and performed 690 assays developed in-house for the molecules.