State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China.
State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China.
Bioorg Chem. 2024 Nov;152:107737. doi: 10.1016/j.bioorg.2024.107737. Epub 2024 Aug 22.
Betulinic acid (BA) is a lupinane-type pentacyclic triterpenoid natural product derived from lupeol that has favorable anti-inflammatory and anti-tumor activities. Currently, BA is mainly produced via botanical extraction, which significantly limits its widespread use. In this study, we investigated the de novo synthesis of BA in Saccharomyces cerevisiae, and to facilitate the synthesis and storage of hydrophobic BA, we adopted a dual-engineering strategy involving peroxisomes and lipid droplets to construct the BA biosynthetic pathway. By expressing Betula platyphylla-derived lupeol C-28 oxidase (BPLO) and Arabidopsis-derived ATR1, we succeeded in developing a BA-producing strain and following multiple expression optimizations of the linker between BPLO and ATR1, the BA titer reached 77.53 mg/L in shake flasks and subsequently reached 205.74 mg/L via fed-batch fermentation in a 5-L bioreactor. In this study, we developed a feasible approach for the de novo synthesis of BA and its direct precursor lupeol in engineered S. cerevisiae.
白桦脂酸（BA）是一种羽扇豆烷型五环三萜天然产物，来源于羽扇醇，具有良好的抗炎和抗肿瘤活性。目前，BA 主要通过植物提取获得，这极大地限制了其广泛应用。在本研究中，我们研究了在酿酒酵母中从头合成 BA 的方法，为了促进疏水性 BA 的合成和储存，我们采用了涉及过氧化物酶体和脂滴的双重工程策略来构建 BA 生物合成途径。通过表达白桦来源的羽扇醇 C-28 氧化酶（BPLO）和拟南芥来源的 ATR1，我们成功开发了一株能够生产 BA 的菌株，并通过对 BPLO 和 ATR1 之间连接肽的多次表达优化，摇瓶中 BA 的产量达到 77.53mg/L，随后通过 5L 生物反应器中的分批补料发酵达到 205.74mg/L。在本研究中，我们开发了一种在工程化酿酒酵母中从头合成 BA 及其直接前体羽扇醇的可行方法。
