Continuous culture of anaerobic fungi enables growth and metabolic flux tuning without use of genetic tools

Leggieri, Patrick A.; Blair, Elaina M.; Lankiewicz, Thomas S.; O'Malley, Michelle A.

doi:10.1016/j.biortech.2023.129854

Title: Continuous culture of anaerobic fungi enables growth and metabolic flux tuning without use of genetic tools

Journal Article · 18 October 2023 · Bioresource Technology

DOI: https://doi.org/10.1016/j.biortech.2023.129854 · OSTI ID:2203000

^[1]; Blair, Elaina M. ^[1]; Lankiewicz, Thomas S. ^[2]; O'Malley, Michelle A. ^[2]

University of California, Santa Barbara, CA (United States)
University of California, Santa Barbara, CA (United States); Joint BioEnergy Institute (JBEI), Emeryville, CA (United States)

Anaerobic gut fungi (AGF) have potential to valorize lignocellulosic biomass owing to their diverse repertoire of carbohydrate-active enzymes (CAZymes). However, AGF metabolism is poorly understood, and no stable genetic tools are available to manipulate growth and metabolic flux to enhance production of specific targets, e.g., cells, CAZymes, or metabolites. Herein, a cost-effective, Arduino-based, continuous-flow anaerobic bioreactor with online optical density control is presented to probe metabolism and predictably tune fluxes in Caecomyces churrovis. Varying the C. churrovis turbidostat setpoint titer reliably controlled growth rate (from 0.04 to 0.20 h⁻¹), metabolic flux, and production rates of acetate, formate, lactate, and ethanol. Bioreactor setpoints to maximize production of each product were identified, and all continuous production rates significantly exceed batch rates. Formate spike-ins increased lactate flux and decreased acetate, ethanol, and formate fluxes. The bioreactor and turbidostat culture schemes demonstrated here offer tools to tailor AGF fermentations to application-specific hydrolysate product profiles.

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Research Organization:: Univ. of California, Oakland, CA (United States); Univ. of California, Santa Barbara, CA (United States); University of California, Santa Barbara, CA (United States)

Sponsoring Organization:: US Army Research Office; USDOE Office of Science (SC); USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231; SC0020420; SC0022142

OSTI ID:: 2203000

Journal Information:: Bioresource Technology, Journal Name: Bioresource Technology Vol. 391; ISSN 0960-8524

Publisher:: Elsevier BVCopyright Statement

Country of Publication:: United States

Language:: English

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