One of the optimization strategies of an artificial biosynthetic metabolic flux with a multienzyme pathway is when the enzyme concentrations are present at the appropriate ratios rather than at their maximum expression. Thus, many recent research efforts have focused on the development of tools that fine-tune the enzyme expression, and these research efforts have facilitated the search for the optimum balance between pathway expression and cell viability. However, the rational approach has some limitations in finding the most optimized expression ratio in in vivo systems. In our study, we focused on fine-tuning the expression level of a six-enzyme reaction for the artificial biosynthesis of curcumin by screening a library of 5′-untranslational region (UTR) sequence mutants made by a multiplex automatic genome engineering (MAGE) tool. From the screening results, a variant (6M08rv) showed about a 38.2-fold improvement in the production of curcumin compared to the parent strain, in which the calculated expression levels of 4-coumarate:CoA ligase (4CL) and phenyldiketide-CoA synthase (DCS), two of the six enzymes, were much lower than those of the parent strain.

中文翻译：

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通过控制5'-UTR随机序列优化大肠杆菌中人工姜黄素的生物合成

具有多酶途径的人工生物合成代谢通量的优化策略之一是当酶浓度以合适的比例而不是其最大表达量存在时。因此，许多最近的研究工作集中在微调酶表达的工具的开发上，这些研究工作促进了途径表达与细胞生存力之间的最佳平衡的寻找。但是，合理的方法在体内寻找最优化的表达比例方面有一定的局限性系统。在我们的研究中，我们专注于通过筛选由多重自动基因组工程（MAGE）制成的5'-非翻译区（UTR）序列突变体的文库，微调用于姜黄素人工生物合成的六酶反应的表达水平。工具。从筛选结果来看，变体（6M08rv）与亲本菌株相比，姜黄素的产量提高了约38.2倍，在亲本菌株中，计算出的4-香豆酸酯：CoA连接酶（4CL）和苯基二酮-CoA合酶的表达水平（六种酶中的两种酶（DCS）远低于亲本菌株。