近日，北京理工大學生命學院高海·軍課題組劉鑫老師，使用IPHASE品牌產品：NADPH再生系統在《Microbial Cell Factories》權·威期刊上發表文章《Engineering cascade biocatalysis in whole cells for syringic acid bioproduction》，影響因子6.4！
丁香酸(Syringic acid, SA)是一種具有多種生物活性的高價值天然化合物，廣泛存在于水果、蔬菜和草藥中。SA主要是通過化學合成來生產的，然而這些化學方法有許多缺點，如設備要求高、反應條件苛刻、催化劑昂貴、副產品眾多等。因此，在本研究中，設計并開發了一種新的利用工程全細胞生產SA的生物轉化途徑。
研究設計了一個多酶級聯體系，在大腸桿菌中利用靜息或生長的全細胞合成SA。本工作鑒定了一種O-甲基化酶(DesAOMT)，它可以催化GA甲基化生成SA。含4種酶的多酶級聯反應在工程大腸桿菌中表達。菌株的代謝系統和生物轉化條件影響催化效率。本研究為SA生物合成提供了一條新的綠色途徑。
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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