Characterization of C-1027 Enediyne Polyketide Synthase

C-1027 烯二炔聚酮合酶的表征

• 批准号: 7055294
• 负责人: Steven Gary Van Lanen
• 金额: $ 4.88万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7055294
• 关键词: active sites; acyl carrier protein; aminoglycoside antibiotics; enzyme mechanism; nuclear magnetic resonance spectroscopy; other phosphotransferase; point mutation; polyketide synthase; postdoctoral investigator; site directed mutagenesis

DESCRIPTION (provided by applicant): The goal of the proposed research is to study the functionality of the domains of the iterative type I enediyne C-1027 polyketide synthase (PKS). Conserved domains will be analysized by site-directed mutagenesis of the proposed active sites, and the mutants characterized by in vivo analysis. It is hypothesized the PKS contains an unusual acyl carrier protein (ACP) domain that requires a unique phosphopantetheinyl transferase (PPTase), which is proposed to also be a domain found within the PKS. The function of the putative ACP and PPTase domain will be analyzed by a thorough in vivo and in vitro analysis, which includes mutant generation of the proposed active sites. Finally, it is proposed that the regiochemistry of enediyne formation is controlled by the PKS. This will be tested by the creation of hybrid PKSs from C- 1027, which contains a 9-carbon enediyne core, and calicheamicin, which contains a 10-carbon enediyne core. The results will provide insights into the programming rules of enediyne biosynthesisand will allow for the generation of "unnatural" natural enediynes with predicted chemistry.

描述（由申请人提供）：拟议的研究的目的是研究I Enediyne C-1027 Poliketide合酶（PKS）的域域的功能。保守的结构域将通过所提出的活性位点的位置定向诱变进行分析，并以体内分析为特征的突变体。假设PKS包含一个不寻常的酰基载体蛋白（ACP）结构域，该结构域需要独特的磷酸乙基转移酶（PPTase），该酶也提议也是PKS中发现的域。假定的ACP和PPTase结构域的功能将通过彻底的体内和体外分析来分析，其中包括提出的活性位点的突变体产生。最后，有人提出，Enediyne组的区域化学由PK控制。这将通过C-1027的混合PKS创建，其中包含9-碳endiyne核心和Calicheamicin，其中包含10碳Enediyne Core。结果将为Enediyne生物合成的编程规则提供见解，并将允许产生具有预测化学的“不自然”天然Enediynes。

项目成果

Progress in combinatorial biosynthesis for drug discovery.

• DOI: 10.1016/j.ddtec.2006.09.014
• 发表时间: 2006-01-01
• 期刊: Drug discovery today. Technologies
• 作者: Van Lanen, Steven G;Shen, Ben
• 通讯作者: Shen, Ben