Expression-level optimization of a multi-enzyme pathway in the absence of a high-throughput assay.

ABSTRACT: Engineered metabolic pathways often suffer from flux imbalances that can overburden the cell and accumulate intermediate metabolites, resulting in reduced product titers. One way to alleviate such imbalances is to adjust the expression levels of the constituent enzymes using a combinatorial expression library. Typically, this approach requires high-throughput assays, which are unfortunately unavailable for the vast majority of desirable target compounds. To address this, we applied regression modeling to enable expression optimization using only a small number of measurements. We characterized a set of constitutive promoters in Saccharomyces cerevisiae that spanned a wide range of expression and maintained their relative strengths irrespective of the coding sequence. We used a standardized

SUBMITTER: Lee ME 

PROVIDER: S-EPMC3905865 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature