Efficient production of itaconic acid from the single-carbon substrate methanol with engineered Komagataella phaffii.

Background: Amidst the escalating carbon dioxide levels resulting from fossil fuel consumption, there is a pressing need for sustainable, bio-based alternatives to underpin future global economies. Single-carbon feedstocks, derived from CO, represent promising substrates for biotechnological applications. Especially, methanol is gaining prominence for bio-production of commodity chemicals.

Results: In this study, we show the potential of Komagataella phaffii as a production platform for itaconic acid using methanol as the carbon source. Successful integration of heterologous genes from Aspergillus terreus (cadA, mttA and mfsA) alongside fine-tuning of the mfsA gene expression, led to promising initial itaconic acid titers of 28 g·L after 5 days of fed-batch cultivation. Through the combined efforts of process optimization and strain engineering strategies, we further boosted the itaconic acid production reaching titers of 55 g·L after less than 5 days of methanol feed, while increasing the product yield on methanol from 0.06 g·g to 0.24 g·g.

Conclusion: Our results highlight the potential of K. phaffii as a methanol-based platform organism for sustainable biochemical production.