Identification of uncharacterized enzymes using multiplexed measurement of biochemical activities

点击翻译按钮获取中文摘要

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.