Advances in 13C metabolic flux analysis: COMPLETE-MFA

Measuring fluxes by 13C metabolic flux analysis (13C-MFA) has become a key activity in metabolic engineering, biotechnology and medicine. In this talk, I will present a new major advance in the field of 13C-MFA, termed COMPLETE-MFA, that is extending the range of biological systems that can be analyzed and dramatically improves the quality of flux results. COMPLETE-MFA, short for complementary parallel labeling experiments technique for metabolic flux analysis, increases not only the precision but also accuracy of flux results by up to an order-of-magnitude compared to traditional approaches. Practically, COMPLETE-MFA is based on combined analysis of multiple isotopic labeling experiments, where the synergy of using complementary tracers helps in resolving difficult to quantify fluxes. In this talk, I will demonstrate several applications of the COMPLETE-MFA approach using all singly labeled glucose tracers, [1-13C], [2-13C], [3-13C], [4-13C], [5-13C], and [6-13C]glucose to determine precise metabolic fluxes for E. coli, Thermus thermophilus, and yeast. Additionally, I will demonstrate the application of all singly labeled xylose tracers, [1-13C], [2-13C], [3-13C], [4-13C], and [5-13C]xylose to determine precise metabolic fluxes in a highly thermophilic Geobacillus strain. I will conclude by highlighting important new insights that were obtained via COMPLETE-MFA.