近日，北京理工大学生命学院高海军课题组刘鑫老师，使用IPHASE品牌产品：NADPH再生系统在《Microbial Cell Factories》权威期刊上发表文章《Engineering cascade biocatalysis in whole cells for syringic acid bioproduction》，影响因子6.4！
丁香酸(Syringic acid, SA)是一种具有多种生物活性的高价值天然化合物，广泛存在于水果、蔬菜和草药中。SA主要是通过化学合成来生产的，然而这些化学方法有许多缺点，如设备要求高、反应条件苛刻、催化剂昂贵、副产品众多等。因此，在本研究中，设计并开发了一种新的利用工程全细胞生产SA的生物转化途径。
研究设计了一个多酶级联体系，在大肠杆菌中利用静息或生长的全细胞合成SA。本工作鉴定了一种O-甲基化酶(DesAOMT)，它可以催化GA甲基化生成SA。含4种酶的多酶级联反应在工程大肠杆菌中表达。菌株的代谢系统和生物转化条件影响催化效率。本研究为SA生物合成提供了一条新的绿色途径。
摘要
Background Syringic acid (SA) is a high-value natural compound with diverse biological activities and wide applications, commonly found in fruits, vegetables, and herbs. SA is primarily produced through chemical synthesis, nonetheless, these chemical methods have many drawbacks, such as considerable equipment requirements, harsh reaction conditions, expensive catalysts, and numerous by-products. Therefore, in this study, a novel biotransformation route for SA production was designed and developed by using engineered whole cells.
Results An O-methyltransferase from Desulfuromonas acetoxidans (DesAOMT), which preferentially catalyzes a methyl transfer reaction on the meta-hydroxyl group of catechol analogues, was identifed. The whole cells expressing DesAOMT can transform gallic acid (GA) into SA when S-adenosyl methionine (SAM) is used as a methyl donor. We constructed a multi-enzyme cascade reaction in Escherichia coli, containing an endogenous shikimate kinase (AroL) and a chorismate lyase (UbiC), along with a p-hydroxybenzoate hydroxylase mutant (PobA**) from Pseudomonas fuorescens, and DesAOMT; SA was biosynthesized from shikimic acid (SHA) by using whole cells catalysis. The metabolic system of chassis cells also afected the efciency of SA biosynthesis, blocking the chorismate metabolism pathway improved SA production. When the supply of the cofactor NADPH was optimized, the titer of SA reached 133 μM (26.2 mg/L).
Conclusion Overall, we designed a multi-enzyme cascade in E. coli for SA biosynthesis by using resting or growing whole cells. This work identifed an O-methyltransferase (DesAOMT), which can catalyze the methylation of GA to produce SA. The multi-enzyme cascade containing four enzymesexpressed in an engineered E. coli for synthesizing of SA from SHA. The metabolic system of the strain and biotransformation conditions infuenced catalytic efciency. This study provides a new green route for SA biosynthesis.
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