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一个酵母细胞工厂的建设和优化,用于生产活性非自然的金色化物3β-O-Glc2-DM

Yanxin Li¹, Xiaoyan Sun¹, Yanxin Liu²

¹State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.

ACS synthetic biology

|October 30, 2024

概括

我们设计了一个酵母细胞工厂来生产3-O-(β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl) -dammar-24-ene-3β,20S-diol (3β-O-Glc2-DM),这是一个新型的金氏化物前体. 这种化合物在体外表现出心脏保护作用,表明其作为治疗剂的潜力.

科学领域:

代谢工程是代谢工程.
合成生物学合成生物学
药理学药理学是指药理学的学科.

背景情况:

帕纳克斯人参中的金色化物是关键的生物活性化合物,通常在特定的基位上糖化.
作为金斯化物前体的dammaranediol-II (DM) 的葡萄糖类的特征很差,它们的生物活性在很大程度上尚未被探索.
人参酸的结构变化会影响其多样化的药理功能.

研究的目的:

建立一个细胞工厂,用于新生生物合成3-O-(β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl) -dammar-24-ene-3β,20S-diol (3β-O-Glc2-DM).
使用代谢工程策略优化3β-O-Glc2-DM的产生.
评估3β-O-Glc2-DM的潜在心脏保护活性.

主要方法:

使用CRISPR/Cas9进行3β-O-Glc2-DM. de novo生物合成的Saccharomyces cerevisiae细胞工厂的建设.
引入编码dammaranediol-II合成酶和特定UDP-glycosyltransferases (UGTs) 的基因.
代谢工程策略,包括酶过度表达,通路流量平衡和内质网膜工程.
在3L生物反应器中进行料批发发酵,用于大规模生产.

主要成果:

成功建造了一个用于3β-O-Glc2-DM生物合成的酵母细胞工厂.
通过代谢工程将3β-O-Glc2-DM标位从18.9升至148.0毫克/升.
通过料批发发酵实现了766.3 mg/L的最终标位.
在实验室中证明了3β-O-Glc2-DM对H/R诱导的心肌细胞损伤的显著保护作用.

结论:

开发了一个强大而高效的细胞工厂,用于微生物生产3β-O-Glc2-DM.
优化发酵过程显著提高了化合物产量.
确定了3β-O-Glc2-DM作为一个有前途的心脏保护药物候选者.

关键词:

这就是dammarenediol-II的原因.人参酸enoside的使用.代谢工程是代谢工程.合成生物学合成生物学酵母细胞工厂是一个酵母细胞工厂.

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