Construction of lignan glycosides biosynthetic network in Escherichia coli using mutltienzyme modules

作者：Yuqi Qiao, Doudou Huang, Yajing Li, Songfan Jiang, Xiao Chen, Junfeng Chen, Ying Xiao, Wansheng Chen · 发表于：Microbial Cell Factories · 年份：2024 · DOI：10.1186/s12934-024-02467-1 · 被引用次数：6 · 研究领域：Plant-derived Lignans Synthesis and Bioactivity、Phytoestrogen effects and research、Phytochemistry and Biological Activities

BACKGROUND: Due to the complexity of the metabolic pathway network of active ingredients, precise targeted synthesis of any active ingredient on a synthetic network is a huge challenge. Based on a complete analysis of the active ingredient pathway in a species, this goal can be achieved by elucidating the functional differences of each enzyme in the pathway and achieving this goal through different combinations. Lignans are a class of phytoestrogens that are present abundantly in plants and play a role in various physiological activities of plants due to their structural diversity. In addition, lignans offer various medicinal benefits to humans. Despite their value, the low concentration of lignans in plants limits their extraction and utilization. Recently, synthetic biology approaches have been explored for lignan production, but achieving the synthesis of most lignans, especially the more valuable lignan glycosides, across the entire synthetic network remains incomplete. RESULTS: By evaluating various gene construction methods and sequences, we determined that the pCDF-Duet-Prx02-PsVAO gene construction was the most effective for the production of (+)-pinoresinol, yielding up to 698.9 mg/L after shake-flask fermentation. Based on the stable production of (+)-pinoresinol, we synthesized downstream metabolites in vivo. By comparing different fermentation methods, including "one-cell, one-pot" and "multicellular one-pot", we determined that the "multicellular one-pot" method ...