Collaborative Research: Genomic Approaches to Metabolic Engineering of Solventogenic Clostridia

Papoutsakis 0418289该提案的目的是使用DNA阵列和“系统生物学”方法来分析和产生适合于连续或补料分批生物处理的无孢子、溶剂产生菌株，以及用于研究大细胞程序（孢子形成）和表型（溶剂生成和退化）。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。这些研究将辅之以发酵和显微镜研究，以捕捉各种菌株的分化过程。一个具有基础和实际意义的重要目的是确定pSOL 1-megaplasm上负责在pSOL 1丧失时抑制孢子形成的一个或多个基因。还将采用一种独立的策略，以产生无孢子但产溶剂的菌株。第三个目的是开发一个关键的遗传工具，代谢工程（ME）的合成和功能基因组分析，即一个有效的重组系统的染色体整合和基因缺失的基础上先进的遗传机制，如组II内含子技术。第四个目标是通过采用几种遗传和ME工具，开发在溶剂生产和有效生物加工方面具有新的和理想特性的菌株。该项目为学生在先进的跨学科研究环境中快速发展的基因组学和ME领域的教育和培训提供了独特的机会。

Papoutsakis0418289The 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.