Characterization and Engineering of Fungal Megasynthases

真菌大合成酶的表征和工程

基本信息

批准号：
8991320
负责人：
Yi Tang
金额：
$ 33.76万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-08-01 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8991320
关键词：
Acetyl-CoA C-Acetyltransferase Acyl Carrier Protein Acyltransferase Address Aflatoxins Anabolism Aspergillus Bacterial Typing Biochemical Biochemistry Biodiversity Biological Biological Models Chemicals Cholesterol Classification Complex Computer Simulation Cyclization Cytochalasins Diels Alder reaction Engineering Enzymatic Biochemistry Enzymes Evolution Family Family member Fingerprint Generations Genes Genome Goals Grant Health Individual Knowledge Lead Learning Link Lovastatin Metabolism Methods Mining Modeling Modification Mycotoxins Natural Products Pathway interactions Penicillium Pharmaceutical Preparations Phylogenetic Analysis Production Reaction Reagent Research Proposals Specificity Structure System Tertiary Protein Structure Type I Polyketide Synthase Work base chemical synthesis cycloaddition decalin drug discovery enzyme pathway experience frontier fungal genetics fungus genome sequencing grasp hypothemycin innovation insight interest microbial polyketide synthase programs protein function protein protein interaction research study tool

项目摘要

DESCRIPTION (provided by applicant): Polyketide synthases (PKSs) of fungal origin are among the most enigmatic enzyme machineries known in microbial secondary metabolism. These enzymes are associated with the biosynthesis of very important fungal natural products, both beneficial, such as the cholesterol lowering blockbuster drug lovastatin from Aspergillus terreus; and deleterious, such as the carcinogenic mycotoxin aflatoxin from different aspergilli. The highly reducing fungal PKSs (HRPKS) are architecturally and mechanistically distinct from the well-characterized bacterial PKSs, and therefore produce compounds of different structural diversity and biological activities. In contrast to bacterial Type I PKSs or the structurally relatd mammalian FASs, our knowledge of the programming rules of fungal HRPKSs remains very limited, especially with regard to how a single set of domains are iteratively used and precisely orchestrated in the synthesis of the final product, often exceeding a combined 30 catalytic steps. Therefore, understanding fungal HRPKS function will enhance our knowledge of natural product biosynthesis and enzymology of multidomain, processive systems. In this proposal, we will comprehensively investigate the mechanisms of HRPKSs using several model systems and compounds. Our overarching goal is to understand, and to be able to predict, the relationship between HRPKS sequence and product structure. Four aims will be pursued in parallel during the proposal period: 1) Classifying and mining fungal HRPKS based on function and product structure; 2) Fingerprinting the specificity of HRPKS tailoring domains; 3) Understanding protein-protein interactions in HRPKS off-loading mechanisms; and 4) Elucidating the enzymes responsible for post- PKS intramolecular cyclization reactions. Each aim is targeted at one unique aspect of HRPKS function that is vital to structural diversity generation. Fundamental enzymology questions will also be answered in addressing these aims. Our work tackles the least understood, the most difficult and perhaps the last frontier of polyketide biosynthesis.

描述（由申请人提供）：真菌起源的聚酮化合物合酶（PKS）是微生物二级代谢中已知的最神秘的酶机之一。这些酶与非常重要的真菌天然产物的生物合成有关，既有有益的，例如降低了曲霉的胆固醇降低了重磅药物lovastatin；和有害的，例如来自不同曲霉的致癌霉菌毒素黄曲霉毒素。高度还原的真菌PKS（HRPK）在结构和机械上与特征良好的细菌PKS不同，因此产生了不同结构多样性和生物学活性的化合物。与细菌I型PKS或结构上相关的哺乳动物FASS相反，我们对真菌HRPKS编程规则的了解仍然非常有限，尤其是关于如何迭代使用并在最终产品合成中精确地精心策划的一组域，通常超过30个催化步骤。因此，了解真菌HRPK功能将增强我们对多域，过程系统的自然产物生物合成和酶学的了解。在此提案中，我们将使用多种模型系统和化合物全面研究HRPKS的机制。我们的总体目标是了解HRPK序列和产品结构之间的关系，并能够预测。在提案期间将同时追求四个目标：1）基于功能和产品结构的真菌HRPK分类和采矿； 2）指纹识别HRPK裁缝域的特异性； 3）了解HRPK中蛋白质 - 蛋白质相互作用的卸载机制； 4）阐明负责后PKS分子内环化反应的酶。每个目标都针对HRPK函数的一个独特方面，这对于结构多样性至关重要。在解决这些目标时，也将回答基本的酶学问题。我们的作品解决了最不理理解的，也许是最困难的，也许是Polyketide Biosynesis的最后一个边界。

项目成果

期刊论文数量（31）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Methylation-dependent acyl transfer between polyketide synthase and nonribosomal peptide synthetase modules in fungal natural product biosynthesis.

DOI：
10.1021/ol503179v
发表时间：
2014-12-19
期刊：
ORGANIC LETTERS
影响因子：
5.2
作者：
Zou, Yi;Xu, Wei;Tsunernatsu, Yuta;Tang, Mancheng;Watanabe, Kenji;Tang, Yi
通讯作者：
Tang, Yi

A fungal ketoreductase domain that displays substrate-dependent stereospecificity.

DOI：
10.1038/nchembio.912
发表时间：
2012-03-11
期刊：
NATURE CHEMICAL BIOLOGY
影响因子：
14.8
作者：
Zhou, Hui;Gao, Zhizeng;Qiao, Kangjian;Wang, Jingjing;Vederas, John C.;Tang, Yi
通讯作者：
Tang, Yi

Comparison of 10,11-Dehydrocurvularin Polyketide Synthases from Alternaria cinerariae and Aspergillus terreus Highlights Key Structural Motifs.