School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China.
Heilongjiang Feihe Dairy Co., Ltd., Beijing 100015, China.
J Agric Food Chem. 2022 Jan 26;70(3):815-825. doi: 10.1021/acs.jafc.1c07035. Epub 2022 Jan 11.
(+)-Nootkatone is an expensive sesquiterpene substance found in grapefruit peels and the heartwood of yellow cedar. It can be used as a food additive, perfume, and insect repellent; therefore, its highly efficient production is greatly needed. However, the low catalytic efficiency of the membrane-anchored cytochrome P450/P450 reductase system (HPO/AtCPR) is the main challenge and limits the production of (+)-nootkatone. We developed an effective high-throughput screening system based on cell wall destruction to probe the optimal ratio of HPO/AtCPR, which achieved a twofold elevation in (+)-valencene oxidation in . An engineered strain PK2RI-AtC/HA was constructed to realize de novo (+)-nootkatone production by a series of metabolic engineering strategies. In biphasic fed-batch fermentation, maximum titers of 3.73 and 1.02 g/L for (+)-valencene and (+)-nootkatone, respectively, were achieved. The dramatically improved performance of the constructed provides an excellent approach for economical production of (+)-nootkatone from glucose.
（+）-诺卡酮是一种从葡萄柚皮和黄杉心材中发现的昂贵的倍半萜物质。它可用作食品添加剂、香料和驱虫剂；因此，非常需要高效生产。然而，膜锚定细胞色素 P450/P450 还原酶系统（HPO/AtCPR）的催化效率低是主要挑战，限制了（+）-诺卡酮的生产。我们开发了一种基于细胞壁破坏的有效高通量筛选系统，以探测 HPO/AtCPR 的最佳比例，在. 中实现了（+）-蒈烯氧化的两倍提高。通过一系列代谢工程策略，构建了一个工程菌株 PK2RI-AtC/HA，实现了从头开始（+）-诺卡酮的生产。在两相补料分批发酵中，分别获得了（+）-蒈烯和（+）-诺卡酮的 3.73 和 1.02 g/L 的最高产量。所构建的 的显著改进性能为从葡萄糖经济生产（+）-诺卡酮提供了一种极好的方法。
