Identification of uncharacterized enzymes using multiplexed measurement of biochemical activities

使用生化活性的多重测量来鉴定未表征的酶

• 批准号: 418754-2012
• 负责人: Caudy, Amy
• 金额: $ 2.04万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2017
• 资助国家: 加拿大
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=630076
• 关键词: Identification; uncharacterized; enzymes; using; multiplexed

Despite nearly two centuries of research, our understanding of metabolic biochemistry is incomplete. The core processes of glycolysis, the TCA cycle, the pentose phosphate pathway, nucleotide synthesis, and amino acid biosynthesis are highly conserved across the three domains of life. This conservation has allowed biochemists to build a metabolic chart incorporating experimental data derived from organisms ranging from E. coli to budding yeast to bovine heart. Construction of a metabolic map using this wide array of model systems has led to an over-generalized understanding so that information about a particular organism may be incomplete or incorrect. In addition to these generalizations and mis-assignments are far more numerous sins of omission in which the gene encoding the enzyme for an anticipated step has not been cloned. These uncharacterized steps in metabolism prevent accurate predictions of biochemical processes, hampering efforts to engineer yeast for purposes of industrial fermentations used to manufacture foodstuffs, biochemicals, and biofuels. The proposed project will scan the genome to identify enzymes catalyzing biochemical activities that have been observed but never have been linked to a gene. This project pioneers a pooling strategy that will allow the efficient screening of the entire yeast genome for enzyme activities of interest.

尽管经过了近两个世纪的研究，我们对代谢生物化学的理解还是不完整的。糖酵解、TCA循环、戊糖磷酸途径、核苷酸合成和氨基酸生物合成的核心过程在生命的三个领域都是高度保守的。这种守恒使生化学家能够建立一个代谢图表，结合从大肠杆菌到出芽酵母再到牛心脏等生物体的实验数据。使用这种广泛的模型系统构建代谢图导致了一种过度一般化的理解，因此关于特定生物体的信息可能是不完整或不正确的。除了这些概括和错误分配之外，还有更多的遗漏罪，其中编码预期步骤的酶的基因没有被克隆出来。这些未被描述的代谢步骤阻碍了对生物化学过程的准确预测，阻碍了为制造食品、生物化学和生物燃料的工业发酵而设计酵母的努力。拟议中的项目将扫描基因组，以确定催化生物化学活动的酶，这些酶已被观察到，但从未与基因联系起来。该项目开创了一个汇集策略，将允许有效筛选整个酵母基因组的酶活性感兴趣。