Synergizing 13C Metabolic Flux Analysis and Metabolic Engineering for Biochemical Production

Abstract

Metabolic engineering of industrial microorganisms to produce chemicals, fuels, and drugs has attracted increasing interest as it provides an environment-friendly and renewable route that does not depend on depleting petroleum sources. However, the microbial metabolism is so complex that metabolic engineering efforts often have difficulty in achieving a satisfactory yield, titer, or productivity of the target chemical. To overcome this challenge, C Metabolic Flux Analysis (C-MFA) has been developed to investigate rigorously the cell metabolism and quantify the carbon flux distribution in central metabolic pathways. In the past decade, C-MFA has been widely used in academic labs and the biotechnology industry to pinpoint the key issues related to microbial-based chemical production and to guide the development of the appropriate metabolic engineering strategies for improving the biochemical production. In this chapter we introduce the basics ofC-MFA and illustrate how C-MFA has been applied to synergize with metabolic engineering to identify and tackle the rate-limiting steps in biochemical production.

¹³C-metabolic flux analysis in glycerol-assimilating strains of <i>Saccharomyces cerevisiae</i>

“…Metabolic flux analysis using 13 C-labeled carbon sources enables the estimation of intracellular metabolic flux distribution and is an effective tool for evaluating the performance of metabolically engineered cells for bioproduction, as well as understanding the mechanism of intracellular metabolism (Antoniewicz, 2015;Guo et al, 2018;Toya and Shimizu, 2013). In metabolic flux analysis, 13 C-labeled carbon sources are incorporated into cells, and enrichment of 13 C to proteinogenic amino acids and/or intracellular metabolites is measured with mass spectrometry.…”