Precision Fermentation as a Frontier in Biofuel Production: Advances, Challenges, and Integration into Biorefineries
2026
Daiane Barão Pereira | Giovanna Lima-Silva | Larissa Batista do Nascimento Soares | Lorena Vieira Bentolila de Aguiar | Aldenora dos Santos Vasconcelos | Vítor Alves Pessoa | Roberta Pozzan | Josilene Lima Serra | Ceci Sales-Campos | Larissa Ramos Chevreuil | Walter José Martínez-Burgos
The industrial transition to advanced biofuels is currently limited by the metabolic constraints and low inhibitor tolerance of wild-type microbial hosts. This review justifies the necessity of Precision Fermentation (PF) as the pivotal technological framework to overcome these barriers, providing a systematic synthesis of high-resolution genetic tools and intelligent bioprocess architectures. We analyze how the integration of CRISPR-Cas9, retron-mediated recombineering, and synthetic regulatory circuits enables the development of specialized microbial “chassis” capable of achieving 10- to 100-fold higher yields compared to native organisms, with industrial titers reaching 50 g/L for isobutanol and 25 g/L for farnesene. A major novelty of this work is the critical evaluation of Artificial Intelligence (AI), Soft Sensing, and Digital Twins in orchestrating real-time metabolic control and mitigating the toxic effects of advanced alcohols and drop-in hydrocarbons (C15–C20). Furthermore, the study concludes that the “scale-out” modular strategy, when integrated into hybrid thermochemical-biochemical biorefineries, allows for the full valorization of C5/C6 sugars and lignin, achieving a Minimum Selling Price (MSP) competitive with fossil fuels. By mapping the synergy between advanced metabolic engineering and data-driven process optimization, this review establishes PF as an indispensable driver for achieving carbon-neutral and carbon-negative energy systems in the circular bioeconomy.
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