Metabolic engineering tools for Saccharomyces cerevisiae: Engineering the spatial organization of metabolic pathways

酿酒酵母的代谢工程工具：代谢途径的空间组织工程

• 批准号: 312357-2012
• 负责人: Martin, Vincent
• 金额: $ 2.48万
• 依托单位: Concordia University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2012
• 资助国家: 加拿大
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=506046
• 关键词: Metabolic; engineering; tools; Saccharomyces; cerevisiae

Much of today's metabolic engineering of S. cerevisiae assumes a simplistic and often naïve approach whereby several proteins, sometimes from widely different origins such as plants, fungi, bacteria or human are collectively expressed in a cell to reconstitute a biochemical pathway. To evaluate the efficacy of the engineered pathways, substrate, product(s) and sometimes intermediates are measured. More often than not, the first generations of these engineered strains show very poor yields and titers and must be optimized for the strains to have any commercial value. Traditionally, the development of each of these strains requires lengthy periods, often consuming over 5-10 years per development cycle. As the pathway complexity goes up so does the time for development. Less than adequate heterologous protein expression or activity is probably the leading cause of poor performance by reconstituted pathways. Other limitations include insufficient supply of precursors, imbalanced pathway leading to reduced flux, accumulation of toxic intermediate(s), loss of intermediate(s) to competing pathway(s) or secretion, aberrant or sub-optimal spatial distribution of recombinant proteins. In this current application, we propose to address the latter.

当今酿酒酵母的大部分代谢工程都采用一种简单且常常天真的方法，即几种蛋白质（有时来自植物、真菌、细菌或人类等不同来源）在细胞中共同表达，以重建生化途径。为了评估工程化途径的功效，需要测量底物、产物，有时还测量中间体。通常，这些工程菌株的第一代产量和滴度非常差，必须对菌株进行优化才能具有商业价值。传统上，每种菌株的开发都需要很长的时间，每个开发周期通常需要 5-10 年以上。随着途径复杂性的增加，开发时间也会增加。异源蛋白表达或活性不足可能是重构途径性能不佳的主要原因。其他限制包括前体供应不足、导致流量减少的不平衡途径、有毒中间体的积累、竞争途径或分泌的中间体损失、重组蛋白的异常或次优空间分布。在当前的应用程序中，我们建议解决后者。