Platforms for nonribosomal peptide manipulation(RMI)

非核糖体肽操作（RMI）平台

• 批准号: 7011324
• 负责人: CLAY CC WANG
• 金额: $ 28.53万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-23 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7011324
• 关键词: Escherichia coli; analog; bacterial genetics; biotechnology; biotherapeutic agent; gene expression; genetic manipulation; synthetic peptide; transfection /expression vector

DESCRIPTION (provided by applicant): The goal of this three year proposal is to develop E. coli as an efficient heterologous expression system for the DNA bisintercalator triostin. A non-ribosomally synthesized peptide, triostin is a known antibiotic and anticancer agent. We will test the hypothesis that by metabolically engineering the E. coli host, modifying the plasmid vector, and altering the triostin gene, triostin and its analogs could be biosynthesized. There are two specific aims for this research, (1) Metabolically engineer E. coli to produce triostin. (2) Modify the triostin NRPS to produce analogs with altered sequence specificity and selectivity. Preliminary results. We have identified a four module non-ribosomal peptide synthetase that contains adenylation domains specific for Serine, Alanine, Cysteine, Valine amino acids (the four amino acids found in triostin.) The significance of this research is that it will (a) improve the mechanistic understanding of the assembly of C2-symmetric non-ribosomal peptides, (b) provide a general host for the expression of diverse non-ribosomal peptide synthetase genes.

描述（由申请人提供）：本三年计划的目标是开发E。大肠杆菌作为一个有效的异源表达系统的DNA双插层triostin。一种非核糖体合成的肽，曲骨素是已知的抗生素和抗癌剂。我们将测试的假设，通过代谢工程的E。coli宿主，改造质粒载体，改变triostin基因，可以生物合成triostin及其类似物。本研究有两个具体的目的：（1）代谢工程E。大肠杆菌中产生曲菌素。(2)修饰曲骨素NRPS以产生具有改变的序列特异性和选择性的类似物。初步结果。我们已经鉴定了一种四模块非核糖体肽合成酶，其含有对丝氨酸、丙氨酸、半胱氨酸、缬氨酸氨基酸（在曲骨素中发现的四种氨基酸）特异性的腺苷酸化结构域。本研究的意义在于：（a）提高了对C2对称非核糖体肽组装机制的理解;（B）为多种非核糖体肽合成酶基因的表达提供了通用宿主。