大肠杆菌中利用甲羟戊酸途径和二萜合成酶的组合工程用于顺式冷杉醇的生产

利用合成生物学技术鉴定二萜合酶编码基因为顺式冷杉醇的生物合成提供了机会。甲基赤藓糖醇磷酸（MEP）和甲羟戊酸（MVA）途径均被设计用于大肠杆菌中顺式冷杉醇的生产，与单独过表达MEP途径或原始MEP途径获得的产量相比，顺式冷杉醇产量分别提高了约7倍和31倍。此外，还对顺式冷杉醇的生物合成进行了系统优化，如二萜合酶的筛选和两相培养。研究发现，来自冷杉的双功能I/II类顺式冷杉醇合酶（AbCAS）和来自鼠尾草的II类冷杉醇合酶（SsTPS2）的组合最为有效。以肉豆蔻酸异丙酯为溶剂，在补料分批生物反应器中进行两相培养，顺式冷杉醇产量达到634.7 mg/L。

Identifying diterpene synthase-encoding genes via synthetic biology technologies provides opportunities for the biosynthesis of cis-abienol. Both the methylerythritol phosphate (MEP) and mevalonate (MVA) pathways were engineered for cis-abienol production in Escherichia coli. Compared with the yields obtained by solely overexpressing the MEP pathway or the native MEP pathway, the cis-abienol titer was improved by approximately 7-fold and 31-fold, respectively. Furthermore, systematic optimization of cis-abienol biosynthesis was conducted, including diterpene synthase screening and two-phase cultivation. It was found that the combination of the bifunctional class I/class II cis-abienol synthase (AbCAS) from Abies and the class II abienol synthase (SsTPS2) from Salvia exhibited the highest efficiency. When two-phase cultivation was performed in a fed-batch bioreactor using isopropyl myristate as the solvent, the cis-abienol titer reached 634.7 mg/L.

搜集汇总

背景与挑战

背景概述

该数据集聚焦于通过合成生物学技术，在大肠杆菌中组合工程甲羟戊酸途径和二萜合成酶以生产顺式冷杉醇。研究通过优化MEP和MVA途径，并筛选高效二萜合酶（如AbCAS和SsTPS2），结合两相培养策略，使顺式冷杉醇产量显著提升，最终在补料分批生物反应器中达到634.7 mg/L。