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Engineering Carotenoid Derived Aroma Compound Pathways into E. coli by Exploring the Sequence Diversity of Putative Carotenoid Dioxygenases Present in Microbial Genome Sequences

通过探索微生物基因组序列中存在的假定类胡萝卜素双加氧酶的序列多样性，将类胡萝卜素衍生的芳香化合物途径工程化到大肠杆菌中

基本信息

批准号：
0332478
负责人：
Claudia Schmidt-Dannert
金额：
$ 32.09万
依托单位：
University of Minnesota-Twin Cities
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
2003
资助国家：
美国
起止时间：
2003-11-01 至 2007-10-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=0332478&HistoricalAwards=false
关键词：
Engineering Carotenoid Derived Aroma Compound

项目摘要

Schmidt-Dannert 0332478The vast number of new gene sequences generated by genome sequencing projects provides an enormous opportunity for the design of new pathways in the post-genomic area. Yet uncharacterized sequence homologs to known biosynthetic enzymes could be used for in vitro recombination with the characterized homologs to access a much larger sequence space (as opposed to using only characterized sequences) for the discovery of new metabolic pathways in such created libraries. This application seeks to expand on the unique set of novel carotenoid pathways available to create for the first time new biosynthetic routes in E. coli for the synthesis of carotenoid derived aroma and flavor compounds. By extending previously engineered carotenoid pathways with novel, uncharacterized bacterial CCD homologs identified in genome sequences, novel biosynthetic pathways for the production of a diverse array of natural and non-natural aroma and color compounds will be generated. Comparison of substrate specificities and cleavage products of CCD homologs will improve our understanding of the mechanistic aspects of carotenoid dioxygenase cleavage. To access the sequences diversity presented by numerous uncharacterized gene homologs identified in genome sequences for in vitro evolution of novel catalytic functions, a novel sequence guided in vitro recombination approach allowing recombination of a large number of fairly diverse sequences will be developed.

由基因组测序计划产生的大量新基因序列为后基因组领域的新途径设计提供了巨大的机会。然而，已知生物合成酶的未表征的序列同源物可用于与表征的同源物的体外重组，以获得更大的序列空间（与仅使用表征的序列相反），用于在这样创建的文库中发现新的代谢途径。本申请旨在扩大独特的一套新的类胡萝卜素途径，首次在大肠杆菌中创造新的生物合成途径。大肠杆菌用于合成类胡萝卜素衍生的香味和风味化合物。通过用在基因组序列中鉴定的新的、未表征的细菌CCD同系物扩展先前工程化的类胡萝卜素途径，将产生用于生产多种天然和非天然芳香和颜色化合物的新的生物合成途径。CCD同源物的底物特异性和裂解产物的比较将提高我们对类胡萝卜素双加氧酶裂解机制方面的理解。为了获得由基因组序列中鉴定的许多未表征的基因同源物呈现的序列多样性，以用于新催化功能的体外进化，将开发允许大量相当多样化的序列重组的新序列引导的体外重组方法。