对A-74528生物合成途径的分析和重构
Jay T Fitzgerald1, Louise K Charkoudian, Katharine R Watts
1Department of Chemistry, Stanford University, Stanford, California 94305, USA.
Journal of the American Chemical Society
|February 28, 2013
概括
A-74528,是一种2的抑制剂.
科学领域:
- 生物化学 生物化学
- 自然产品 化学 化学
- 分子生物学分子生物学
背景情况:
- A-74528是一种C30多基化物天然产品,可以抑制2',5'-奥利戈亚丁酸二化酶 (2'-PDE).
- 2'-PDE是干扰素通路中的关键酶,使其调节成为病毒感染的潜在治疗策略.
- 对A-74528的生物合成基因集群也产生fredericamycin,这是一个结构上不同的细胞毒剂.
研究的目的:
- 为了确定负责A-74528生物合成的基因.
- 了解A-74528形成背后的酶机制.
- 为了设计一个异质宿主,在没有fredericamycin的情况下高效地生产A-74528.
主要方法:
- 在A-74528生物合成基因集群中的淘汰突变体的构建和分析.
- 编码定制酶 (循环酶,氧化酶) 和核心聚基合成酶的基因的识别.
- 在异质宿主 (Streptomyces coelicolor CH999) 中推断途径的遗传重构.
主要成果:
- 确定了A-74528生物合成所必需的关键基因.
- 包括立体中心和环循环在内的A-74528形成主要由两个定制酶和多基基合成酶催化.
- 重新设计的Streptomyces coelicolor CH999产生了3毫克/升的A-74528,没有弗雷德里卡米辛污染.
结论:
- 已经阐明了A-74528生物合成所需的最小酶机制.
- 基因工程使A-74528在异质宿主中能够有效地产生,为治疗开发铺平了道路.
- 这项工作为生产A-74528和相关化合物用于抗病毒疗法的基础.
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