Yuzawa Satoshi, Zargar Amin, Pang Bo, Katz Leonard, Keasling Jay D
Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United States; Joint BioEnergy Institute, Emeryville, CA, United States.
Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United States; QB3 Institute, University of California, Berkeley, CA, United States.
Methods Enzymol. 2018;608:393-415. doi: 10.1016/bs.mie.2018.04.027. Epub 2018 May 26.
Reduced polyketides are a subclass of natural products that have a variety of medical, veterinary, and agricultural applications and are well known for their structural diversity. Although these compounds do not resemble each other, they are all made by a class of enzymes known as modular polyketide synthases (PKSs). The commonality of PKS domains/modules that compose PKSs and the understanding of the relationship between the sequence of the PKS and the structure of the compound it produces render modular PKSs as excellent targets for engineering to produce novel compounds with predicted structures. Here, we describe experimental protocols and considerations for modular PKS engineering and two case studies to produce commodity chemicals by engineered PKSs.
还原型聚酮化合物是一类天然产物，具有多种医学、兽医学和农业应用，并以其结构多样性而闻名。尽管这些化合物彼此并不相似，但它们都是由一类称为模块化聚酮合酶（PKSs）的酶产生的。构成聚酮合酶的PKS结构域/模块的共性以及对聚酮合酶序列与其所产生化合物结构之间关系的理解，使得模块化聚酮合酶成为工程改造以生产具有预测结构的新型化合物的理想靶点。在这里，我们描述了模块化聚酮合酶工程的实验方案和注意事项，以及两个通过工程化聚酮合酶生产商品化学品的案例研究。
