School of Biological Sciences and Biotechnology, Graduate School, Chonnam National University, Gwangju 61186, Republic of Korea.
Department of Chemistry, Chonnam National University, Gwangju 61186, Republic of Korea.
J Agric Food Chem. 2023 Mar 15;71(10):4328-4336. doi: 10.1021/acs.jafc.2c09108. Epub 2023 Mar 1.
One-carbon chemicals (C 1s) are potential building blocks as they are cheap, sustainable, and abiotic components. Methanol-derived formaldehyde can be another versatile building block for the production of 2-keto-4-hydroxyacid derivatives that can be used for amino acids, hydroxy carboxylic acids, and chiral aldehydes. To produce 2-keto-4-hydroxybutyrate from C 1s in an environment-friendly way, we characterized an aldolase from PAO1 (ADL), which showed much higher catalytic activity in condensing formaldehyde and pyruvate than the reported aldolases. By applying a structure-based rational approach, we found a variant (ADL) that exhibited better catalytic activities than the wild-type enzyme. Next, we constructed a one-pot cascade biocatalyst system by combining ADL and a methanol dehydrogenase (MDH) and, for the first time, effectively produced 2-keto-4-hydroxybutyrate as the main product from pyruvate and methanol via an enzymatic reaction. This simple process applied here will help design a green process for the production of 2-keto-4-hydroxyacid derivatives.
一碳化学品 (C1s) 是潜在的构建模块，因为它们廉价、可持续且是非生物成分。甲醇衍生的甲醛可以作为另一种多功能构建块，用于生产 2-酮-4-羟基酸衍生物，这些衍生物可用于氨基酸、羟基羧酸和手性醛。为了以环保的方式从 C1s 生产 2-酮-4-羟基丁酸，我们对来自 PAO1 的醛缩酶 (ADL) 进行了表征，该酶在缩合甲醛和丙酮酸方面表现出比报道的醛缩酶更高的催化活性。通过应用基于结构的合理方法，我们发现了一种变体 (ADL)，其催化活性优于野生型酶。接下来，我们通过组合 ADL 和甲醇脱氢酶 (MDH) 构建了一锅式级联生物催化剂系统，并首次通过酶反应有效地从丙酮酸和甲醇生产 2-酮-4-羟基丁酸作为主要产物。这里应用的这个简单过程将有助于设计用于生产 2-酮-4-羟基酸衍生物的绿色工艺。
