A miniaturized feedstocks-to-fuels pipeline for screening the efficiency of deconstruction and microbial conversion of lignocellulosic biomass.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Lignin*/metabolism ; Lignin*/chemistry ; Biomass* ; Biofuels*/analysis ; Sorghum*/metabolism; Ionic Liquids/chemistry

Sustainably grown biomass is a promising alternative to produce fuels and chemicals and reduce the dependency on fossil energy sources. However, the efficient conversion of lignocellulosic biomass into biofuels and bioproducts often requires extensive testing of components and reaction conditions used in the pretreatment, saccharification, and bioconversion steps. This restriction can result in a significant and unwieldy number of combinations of biomass types, solvents, microbial strains, and operational parameters that need to be characterized, turning these efforts into a daunting and time-consuming task. Here we developed a high-throughput feedstocks-to-fuels screening platform to address these challenges. The result is a miniaturized semi-automated platform that leverages the capabilities of a solid handling robot, a liquid handling robot, analytical instruments, and a centralized data repository, adapted to operate as an ionic-liquid-based biomass conversion pipeline. The pipeline was tested by using sorghum as feedstock, the biocompatible ionic liquid cholinium phosphate as pretreatment solvent, a "one-pot" process configuration that does not require ionic liquid removal after pretreatment, and an engineered strain of the yeast Rhodosporidium toruloides that produces the jet-fuel precursor bisabolene as a conversion microbe. By the simultaneous processing of 48 samples, we show that this configuration and reaction conditions result in sugar yields (~70%) and bisabolene titers (~1500 mg/L) that are comparable to the efficiencies observed at larger scales but require only a fraction of the time. We expect that this Feedstocks-to-Fuels pipeline will become an effective tool to screen thousands of bioenergy crop and feedstock samples and assist process optimization efforts and the development of predictive deconstruction approaches.
Competing Interests: B.A.S. has a financial interest in Caribou Biofuels, Illium Technologies, and Erg Bio. This does not alter our adherence to PLOS ONE policies on sharing data and materials.
(Copyright: © 2024 Pidatala et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Front Bioeng Biotechnol. 2021 Feb 09;9:612893. (PMID: 33634086)
Bioresour Technol. 2016 Jan;199:21-33. (PMID: 26343573)
ACS Synth Biol. 2017 Dec 15;6(12):2248-2259. (PMID: 28826210)
Front Plant Sci. 2020 Mar 11;11:282. (PMID: 32218797)
Metab Eng. 2021 Jan;63:34-60. (PMID: 33221420)
Ann N Y Acad Sci. 2010 May;1195:28-45. (PMID: 20536815)
Biotechnol Biofuels. 2021 Apr 21;14(1):101. (PMID: 33883010)
Appl Microbiol Biotechnol. 2019 Dec;103(23-24):9305-9320. (PMID: 31707441)
Front Bioeng Biotechnol. 2022 Nov 22;10:1061667. (PMID: 36483774)
Biotechnol Biofuels. 2021 Sep 10;14(1):179. (PMID: 34507592)
Environ Sci Pollut Res Int. 2024 Feb;31(9):12871-12891. (PMID: 38285255)
Curr Opin Biotechnol. 2019 Apr;56:147-155. (PMID: 30529238)
Biotechnol Biofuels. 2021 Nov 20;14(1):217. (PMID: 34801067)

9005-53-2 (Lignin)
11132-73-3 (lignocellulose)
0 (Biofuels)
0 (Ionic Liquids)

Date Created: 20241008 Date Completed: 20241008 Latest Revision: 20241010

20250114

PMC11460671

10.1371/journal.pone.0305336

39378235