Metabolic engineering of Kluyveromyces marxianus for biomass-based applications

Kluyveromyces marxianus ATCC 26,548 was cultivated in aerobic chemostats with [1-¹³C] and [U-¹³C] glucose as carbon source under three different growth conditions (0.10, 0.25, and 0.5 h^-1) to evaluate metabolic fluxes. Carbon balances closed always within 97-102%. Growth was carbon limited, and the cell yield on glucose was the same. The extracellular side-product formation was very low, totaling 0.0008 C-mol C-mol^-1 substrate at 0.5 h^-1. The intracellular flux ratios did not show significant variation for metabolic flux analysis from labelling and biomass composition and metabolic flux ratio analysis from labelling. The observed strictly oxidative metabolism and the stability of the metabolism in terms of fluxes even at high growth rates, without triggering out the synthesis of by-products, is an extremely desired condition that underlines the potential of K. marxianus for biotechnological biomass-related applications and the comprehension of the metabolic pools and pathways is an important step to engineering this organism.

The online version contains supplementary material available at 10.1007/s13205-022-03324-x.