Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
Department of Biological Sciences, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
J Agric Food Chem. 2022 Mar 2;70(8):2656-2663. doi: 10.1021/acs.jafc.1c07398. Epub 2022 Feb 1.
Cinnamaldehyde (CAD) has various applications in foods and pharmaceuticals and has gained prominence as a potent nematicide in agricultural research owing to its nematicidal activity. However, conventional methods of CAD production, including extraction from plants or organic chemical synthesis, are environmentally hazardous and limit its utilization for downstream applications. Here, we engineered as a whole-cell biocatalyst for the efficient bioconversion of -cinnamic acid (-CA) into CAD. An expression module of carboxylic acid reductase was constructed for the conversion of -CA to CAD. Additionally, the putative dehydrogenase-related genes (, , and 1176) responsible for the conversion of CAD to cinnamyl alcohol were deleted from the engineered strain to prevent the loss of CAD. Furthermore, as the conversion is NADPH-dependent, we investigated the conversion efficiency by exchanging the putative promoter region for the gene, which encodes glucose-6-phosphate dehydrogenase, with a strong promoter to increase the NADPH pool. Finally, a bioconversion platform using as a whole-cell biocatalyst was developed by deleting the gene, which is involved in the reverse conversion of CAD to -CA. Taken together, a 100% conversion yield of 1.1 g/L CAD from 1.2 g/L -CA was obtained within 30 min.
肉桂醛（CAD）在食品和药品中有多种应用，由于其杀线虫活性，在农业研究中作为一种有效的杀线虫剂而备受关注。然而，CAD 的传统生产方法，包括从植物中提取或有机化学合成，对环境有害，限制了其在下游应用中的利用。在这里，我们构建了 作为全细胞生物催化剂，用于高效生物转化 -肉桂酸（-CA）为 CAD。构建了表达模块用于将 -CA 转化为 CAD。此外，从工程菌中删除了负责将 CAD 转化为肉桂醇的假定脱氢酶相关基因（,, 和 1176），以防止 CAD 的损失。此外，由于转化是 NADPH 依赖性的，我们通过交换假定的启动子区域用于编码葡萄糖-6-磷酸脱氢酶的 基因，用强启动子增加 NADPH 池，来研究转化效率。最后，通过删除参与 CAD 向 -CA 逆转化的 基因，构建了使用 作为全细胞生物催化剂的生物转化平台。总之，在 30 分钟内从 1.2 g/L -CA 获得了 1.1 g/L CAD 的 100%转化率。
