Advanced one-pot deconstruction and valorization of lignocellulosic biomass into triacetic acid lactone using Rhodosporidium toruloides

Otoupal, Peter B.; Geiselman, Gina M.; Oka, Asun M.; Barcelos, Carolina A.; Choudhary, Hemant; Dinh, Duy; Zhong, Wenqing; Hwang, HeeJin; Keasling, Jay D.; Mukhopadhyay, Aindrila; Sundstrom, Eric; Haushalter, Robert W.; Sun, Ning; Simmons, Blake A.; Gladden, John M.

doi:10.1186/s12934-022-01977-0

Title: Advanced one-pot deconstruction and valorization of lignocellulosic biomass into triacetic acid lactone using Rhodosporidium toruloides

Journal Article · Thu Dec 08 00:00:00 EST 2022 · Microbial Cell Factories

DOI:https://doi.org/10.1186/s12934-022-01977-0· OSTI ID:1902473

Otoupal, Peter B.; Geiselman, Gina M.; Oka, Asun M.; Barcelos, Carolina A.; Choudhary, Hemant; Dinh, Duy; Zhong, Wenqing; Hwang, HeeJin; Keasling, Jay D.; Mukhopadhyay, Aindrila; Sundstrom, Eric; Haushalter, Robert W.; Sun, Ning; Simmons, Blake A.; Gladden, John M.

Abstract Background Rhodosporidium toruloides is capable of co-utilization of complex carbon sources and robust growth from lignocellulosic hydrolysates. This oleaginous yeast is therefore an attractive host for heterologous production of valuable bioproducts at high titers from low-cost, deconstructed biomass in an economically and environmentally sustainable manner. Here we demonstrate this by engineering R. toruloides to produce the polyketide triacetic acid lactone (TAL) directly from unfiltered hydrolysate deconstructed from biomass with minimal unit process operations. Results Introduction of the 2-pyrone synthase gene into R. toruloides enabled the organism to produce 2.4 g/L TAL from simple media or 2.0 g/L from hydrolysate produced from sorghum biomass. Both of these titers are on par with titers from other better-studied microbial hosts after they had been heavily engineered. We next demonstrate that filtered hydrolysates produced from ensiled sorghum are superior to those derived from dried sorghum for TAL production, likely due to the substantial organic acids produced during ensiling. We also demonstrate that the organic acids found in ensiled biomass can be used for direct synthesis of ionic liquids within the biomass pretreatment process, enabling consolidation of unit operations of in-situ ionic liquid synthesis, pretreatment, saccharification, and fermentation into a one-pot, separations-free process. Finally, we demonstrate this consolidation in a 2 L bioreactor using unfiltered hydrolysate, producing 3.9 g/L TAL. Conclusion Many steps involved in deconstructing biomass into fermentable substrate can be combined into a distinct operation, and directly fed to cultures of engineered R. toruloides cultures for subsequent valorization into gram per liter titers of TAL in a cost-effective manner.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Energy Efficiency and Renewable Energy (EERE)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1902473

Alternate ID(s):: OSTI ID: 1958751

Journal Information:: Microbial Cell Factories, Journal Name: Microbial Cell Factories Vol. 21 Journal Issue: 1; ISSN 1475-2859

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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