Collaborative Research: Genomic Approaches to Metabolic Engineering of Solventogenic Clostridia

本提案的目的是使用DNA阵列和“系统生物学”方法来分析和生成适合连续或分批补料生物处理的产孢子、溶剂菌株，并用于研究大型细胞程序（孢子形成）和表型（溶剂发生和退化）。基因组规模的DNA阵列将用于捕捉细胞转录程序的全局图像。这些研究将辅以发酵和显微镜研究，以捕捉各种菌株的分化过程。确定pSOL1巨质粒上在pSOL1丢失时抑制产孢的基因是一个重要的基础和现实意义。还将采取一项独立的战略，以产生产菌种但具有溶剂性的菌株。第三个目标是开发一种用于代谢工程（ME）合成和功能基因组分析的关键遗传工具，即基于先进遗传机制（如II组内含子技术）的染色体整合和基因缺失的有效重组系统。第四个目标是通过使用几种遗传和ME工具，在溶剂生产和高效生物处理方面开发具有新的和理想特性的菌株。该项目为快速发展的基因组学和ME领域的学生提供了独特的教育和培训机会，并提供了先进的跨学科研究环境。

Papoutsakis0418157The aims of this proposal are to use DNA arrays and a "systems biology" approach for the analysis and generation of asporogenous, solvent producing strains suitable for continuous or fed-batch bioprocessing, and for studying large cellular programs (sporulation) and phenotypes (solventogenesis and degeneration). Genomic-scale DNA arrays will be used to capture the global picture of cellular transcriptional programs. These studies will be complemented by fermentation and microscopic studies to capture the differentiation process of the various strains. An important aim of both fundamental and practical significance is to determine the gene or genes on the pSOL1-megaplasmid responsible for inhibiting sporulation upon pSOL1 loss. An independent strategy will be also pursued in order to generate asporogenous but solventogenic strains. A third aim is to develop a critical genetic tool for both metabolic engineering (ME) synthesis and functional-genomic analysis, namely that of an efficient recombination system for chromosomal integration and gene deletion based on advanced genetic mechanisms such as the group II intron technology. A fourth aim is to develop strains with new and desirable characteristics in terms of solvent production and efficient bioprocessing, by employing several genetic and ME tools. This project provides unique opportunities for student education and training in the fast growing fields of genomics and ME in an advanced interdisciplinary research environment.