De novo metabolic engineering of a designed biosynthetic pathway for complex terpenoids in yeast

酵母中复杂萜类化合物设计生物合成途径的从头代谢工程

• 批准号: 5448252
• 负责人: Dr. Stefan Jennewein
• 金额: --
• 依托单位: Fraunhofer-Institut für Molekularbiologie und Angewandte Oekologie (IME)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2005
• 资助国家: 德国
• 起止时间: 2004-12-31 至 2008-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5448252?language=en
• 关键词: De; novo; metabolic; engineering; designed

Engineered microbial systems producing complex natural products would provide an efficient and environmental friendly alternative to chemical synthetic processes or the extraction from natural sources. For many natural products the identification of the underlying biosynthetic pathways mad emajor advances. And the metabolic pathway engineering for several natural products in microbial hosts is now becoming possible. However, the use of the identified biosynthetic genes for such metabolic engineering approaches is often limited, due to incompatible substrate specificity, codon usage patterns, regulation, required substrate pressure etc. In vitro evolution proved to be a powerful tool for the efficient tailoring of biocatalysts. In addition, the use of in vitro evolution will allow to a significant extend the rational design of the engineered biosynthetic pathway. The here proposed project wants to examine the concept of de novo metabolic engineering of rational designed biosynthetic pathways, using tailor-made biocatalysts, on the example of the taxoid diterpenoids. In Taxus the taxoid biosynthetic pathway proved to be an anastamosing Biosynthetic network, with many metabolic diversions, leading to many closely related biosynthetic products. Using the identified biosynthetic genes the project attempts to establish a linear rational designed biosynthetic pathway for defined taxoid products in yeast. This attempt represents a solution for the long-standing problem of adequate supply and sustainable production of the currently used clinically taxanes (Taxol & Taxotere). In addition, the projekt will open new avenues for the synthesis of novel, third generation clinical taxanes with superior biological activity. Besides addressing the metabolic engineering of a highly oxygenated terpenoid, using cytochrome P450 dependent monooxygenases, in particular, the projekt will also deliver new insight into the metabolic engineering of yeast in general. Therefore, the proposed projekt will represent a major advancement in the metabolic engineering of yeast for the use as cell factories for the production of challenging, complex fine chemicals.

生产复杂天然产物的工程微生物系统将为化学合成工艺或从天然来源提取提供有效且环境友好的替代方案。对许多天然产物而言，对潜在的生物合成途径的鉴定是一个重要的进展。同时，利用微生物宿主对天然产物进行代谢途径工程也成为可能。然而，由于不相容的底物特异性，密码子使用模式，调节，所需的底物压力等，使用这种代谢工程方法的生物合成基因往往是有限的。体外进化被证明是一个强大的工具，有效的剪裁生物催化剂。此外，体外进化的使用将允许显著扩展工程化生物合成途径的合理设计。这里提出的项目要检查的概念从头代谢工程的合理设计的生物合成途径，使用量身定制的生物催化剂，紫杉烷类化合物的例子。红豆杉的紫杉烷生物合成途径是一个相互连接的生物合成网络，具有多种代谢转向，产生许多密切相关的生物合成产物。利用已鉴定的生物合成基因，该项目试图建立一个线性的合理设计的生物合成途径，以确定紫杉烷产品在酵母中。这一尝试代表了目前临床使用的紫杉烷（Taxol & Taxotere）的充足供应和可持续生产的长期问题的解决方案。此外，该项目将为合成具有上级生物活性的新型第三代临床紫杉烷开辟新的途径。除了使用细胞色素P450依赖性单加氧酶解决高度氧化的萜类化合物的代谢工程外，该项目还将为酵母的代谢工程提供新的见解。因此，拟议的项目将代表酵母代谢工程的重大进展，用于生产具有挑战性的复杂精细化学品的细胞工厂。