Improved Itaconate Production with Ustilago cynodontis via Co-Metabolism of CO2-Derived Formate
2022
Lena Ullmann | Nils Guntermann | Philipp Kohl | Gereon Schröders | Andreas Müsgens | Giancarlo Franciò | Walter Leitner | Lars M. Blank
In recent years, it was shown that itaconic acid can be produced from glucose with Ustilago strains at up to maximum theoretical yield. The use of acetate and formate as co-feedstocks can boost the efficiency of itaconate production with Ustilaginaceae wild-type strains by reducing the glucose amount and thus the agricultural land required for the biotechnological production of this chemical. Metabolically engineered strains (U. cynodontis &Delta:fuz7 &Delta:cyp3 &uarr:Pria1 and U. cynodontis &Delta:fuz7 &Delta:cyp3 PetefmttA &uarr:Pria1) were applied in itaconate production, obtaining a titer of 56.1 g L&minus:1 and a yield of 0.55 gitaconate per gsubstrate. Both improved titer and yield (increase of 5.2 g L&minus:1 and 0.04 gitaconate per gsubstrate, respectively) were achieved when using sodium formate as an auxiliary substrate. By applying the design-of-experiments (DoE) methodology, cultivation parameters (glucose, sodium formate and ammonium chloride concentrations) were optimized, resulting in two empirical models predicting itaconate titer and yield for U. cynodontis &Delta:fuz7 &Delta:cyp3 PetefmttA &uarr:Pria1. Thereby, an almost doubled itaconate titer of 138 g L&minus:1 was obtained and a yield of 0.62 gitaconate per gsubstrate was reached during confirmation experiments corresponding to 86% of the theoretical maximum. In order to close the carbon cycle by production of the co-feed via a &ldquo:power-to-X&rdquo: route, the biphasic Ru-catalysed hydrogenation of CO2 to formate could be integrated into the bioprocess directly using the obtained aqueous solution of formates as co-feedstock without any purification steps, demonstrating the (bio)compatibility of the two processes.
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