SusChEM: Installing the Wood-Ljungdahl pathway in a clostridium platform organism for enhanced metabolite production

SusChEM：在梭菌平台生物体中安装 Wood-Ljungdahl 途径以增强代谢产物的产生

• 批准号: 1511660
• 负责人: Eleftherios Papoutsakis
• 金额: $ 60万
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-15 至 2018-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1511660&HistoricalAwards=false
• 关键词: SusChEM; Installing; Wood; Ljungdahl; pathway

1511660 Papoutsakis, Eleftherios T. The goal of this project is to engineer the industrial organism Clostridium acetobutylicum to make it capable of utilizing carbon dioxide, carbon monoxide and hydrogen, as sole substrates for growth and product formation. This will enable the production of biofuels (e.g. butanol) and other useful chemicals from waste gases in a fermentation system that can be easily scaled for industrial production, and with exceptional efficiencies. From the fundamental point of view, the ability to install this complex, primordial pathway in a different organism constitutes a major advance in synthetic biology that would open new horizons for pathway engineering and synthetic approaches in other industrial microorganisms. The most efficient pathway for non-photosynthetic chemotrophic carbon utilization is the Wood-Ljungdahl pathway (WLP) employed by acetogens, which are Clostridium organisms that fix enormous quantities of carbon dioxide in the geobiosphere. Comparative genomic analysis of the three sequenced genomes of mesophilic acetogens showed that the WLP genes of all three organisms are concentrated in a highly conserved region in their respective genomes. Based on the published literature and work from the investigator's laboratory, the model acetogen C. ljungdahlii was chosen as the source of most of the WLP genes that will be used to install the WLP in C. acetobutylicum. The plan is to integrate a number of genes into the chromosome and express other genes on two large compatible/co-existing plasmids. Chromosomal integrations will be based on a recently developed technology for double-crossover allelic DNA exchange that enables markerless chromosomal gene deletions and DNA integration. The constructed strains will be assessed for the WLP functionality through in vitro assays, growth experiments and carbon-13 based carbon-fixation assays.This award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biology.

1511660 Papoutsakis，Eleftherios T。该项目的目标是改造工业生物体丙酮丁醇梭菌，使其能够利用二氧化碳、一氧化碳和氢气作为生长和产物形成的唯一底物。 这将使发酵系统中的废气能够生产生物燃料（例如丁醇）和其他有用的化学品，该发酵系统可以轻松地扩大规模用于工业生产，并且效率极高。从基本角度来看，在不同生物体中安装这种复杂的原始途径的能力构成了合成生物学的重大进步，将为其他工业微生物的途径工程和合成方法开辟新的视野。非光合化能碳利用的最有效途径是产乙酸菌所采用的伍德-永达尔途径（WLP），产乙酸菌是在地球生物圈中固定大量二氧化碳的梭状芽孢杆菌生物体。对嗜温产乙酸菌的三个测序基因组进行的比较基因组分析表明，所有三种生物的 WLP 基因都集中在各自基因组的高度保守区域。 根据已发表的文献和研究者实验室的工作，选择模型产乙酸菌 C. ljungdahlii 作为大多数 WLP 基因的来源，这些基因将用于在丙酮丁醇梭菌中安装 WLP。该计划是将许多基因整合到染色体中，并在两个大的兼容/共存质粒上表达其他基因。染色体整合将基于最近开发的双交叉等位基因 DNA 交换技术，该技术能够实现无标记染色体基因删除和 DNA 整合。构建的菌株将通过体外测定、生长实验和基于碳 13 的碳固定测定来评估 WLP 功能。该奖项由 CBET 部门的生物技术和生化工程计划由系统和合成生物学共同资助分子和细胞生物学司的计划。