School of Food Science and Technology, Jiangnan University, 214122 Wuxi, China.
Food Science and Technology Programme, Nanyang Technological University, 62 Nanyang Drive, Singapore 637459, Singapore.
J Agric Food Chem. 2022 Jun 22;70(24):7490-7499. doi: 10.1021/acs.jafc.2c02137. Epub 2022 Jun 1.
Hydroxytyrosol is an olive-derived phenolic compound of increasing commercial interest due to its health-promoting properties. In this study, a high-yield hydroxytyrosol-producing cell factory was established via a comprehensive metabolic engineering scheme. First, biosynthetic pathway of hydroxytyrosol was constructed in yeast by gene screening and overexpression of different phenol hydroxylases, among which paHD (from ) displayed the best catalytic performance. Next, hydroxytyrosol precursor supply was enhanced via a multimodular engineering approach: elimination of tyrosine feedback inhibition through genomic integration of and , construction of an aromatic aldehyde synthase (AAS)-based tyrosine metabolic pathway, and redistribution of metabolic flux between glycolytic pathway and pentose phosphate pathway (PPP) by introducing the exogenous gene . As a result, the titer of hydroxytyrosol was improved by 6.88-fold. Finally, a glucose-responsive dynamic regulation system based on deletion was implemented, resulting in the final hydroxytyrosol yields of 308.65 mg/L and 167.98 mg/g cell mass, the highest known from production in to date.
羟基酪醇是一种橄榄中提取的酚类化合物，由于其具有促进健康的特性，越来越受到商业关注。在这项研究中，通过综合代谢工程方案，建立了一个高产羟基酪醇的细胞工厂。首先，通过基因筛选和过表达不同的酚羟化酶，在酵母中构建了羟基酪醇的生物合成途径，其中 paHD（来自 ）表现出最好的催化性能。接下来，通过多模块工程方法增强羟基酪醇前体的供应：通过基因组整合 和 消除酪氨酸反馈抑制，构建基于芳香醛合酶（AAS）的酪氨酸代谢途径，以及通过引入外源基因 在糖酵解途径和磷酸戊糖途径（PPP）之间重新分配代谢通量。结果，羟基酪醇的产量提高了 6.88 倍。最后，实施了基于 缺失的葡萄糖响应动态调控系统，最终羟基酪醇的产量达到 308.65mg/L 和 167.98mg/g 细胞干重，这是迄今为止从 生产中获得的最高产量。
