Engineered Pseudomonas putida simultaneously catabolizes five major components of corn stover lignocellulose: Glucose, xylose, arabinose, p-coumaric acid, and acetic acid

Elmore, Joshua R.; Dexter, Gara N.; Salvachúa, Davinia; O'Brien, Marykate; Klingeman, Dawn M.; Gorday, Kent; Michener, Joshua K.; Peterson, Darren J.; Beckham, Gregg T.; Guss, Adam M.

doi:10.1016/j.ymben.2020.08.001

Title: Engineered Pseudomonas putida simultaneously catabolizes five major components of corn stover lignocellulose: Glucose, xylose, arabinose, p-coumaric acid, and acetic acid

Journal Article · Sun Nov 01 00:00:00 EDT 2020 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2020.08.001· OSTI ID:1648386

Elmore, Joshua R.;

; Salvachúa, Davinia; O'Brien, Marykate;

;

; Michener, Joshua K.; Peterson, Darren J.; Beckham, Gregg T.;

Valorization of all major lignocellulose components, including lignin, cellulose, and hemicellulose is critical for an economically viable bioeconomy. In most biochemical conversion approaches, the standard process separately upgrades sugar hydrolysates and lignin. Here, we present a new process concept based on an engineered microbe that could enable simultaneous upgrading of all lignocellulose streams, which has the ultimate potential to reduce capital cost and enable new metabolic engineering strategies. Pseudomonas putida is a robust microorganism capable of natively catabolizing aromatics, organic acids, and D-glucose. We engineered this strain to utilize D-xylose by tuning expression of a heterologous D-xylose transporter, catabolic genes xylAB, and pentose phosphate pathway (PPP) genes tal-tkt. We further engineered L-arabinose utilization via the PPP or an oxidative pathway. This resulted in a growth rate on xylose and arabinose of 0.32 h^-1 and 0.38 h^-1, respectively. Using the oxidative L-arabinose pathway with the PPP xylose pathway enabled D-glucose, D-xylose, and L-arabinose co-utilization in minimal medium using model compounds as well as real corn stover hydrolysate, with a maximum hydrolysate sugar consumption rate of 3.3 g/L/h. After modifying catabolite repression, our engineered P. putida simultaneously co-utilized five representative compounds from cellulose (D-glucose), hemicellulose (D-xylose, L-arabinose, and acetic acid), and lignin-related compounds (p-coumarate), demonstrating the feasibility of simultaneously upgrading total lignocellulosic biomass to value-added chemicals.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States); Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE); USDOE Office of Science (SC), Workforce Development for Teachers and Scientists (WDTS); USDOE Laboratory Directed Research and Development (LDRD) Program

Grant/Contract Number:: 7866; AC36-08GO28308; AC05-00OR22725; AC05-76RLO1830

OSTI ID:: 1648386

Alternate ID(s):: OSTI ID: 1660263; OSTI ID: 1661258; OSTI ID: 1665827

Report Number(s):: NREL/JA-2800-77802; S109671762030121X; PII: S109671762030121X

Journal Information:: Metabolic Engineering, Journal Name: Metabolic Engineering Vol. 62 Journal Issue: C; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: Belgium

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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Title: Engineered Pseudomonas putida simultaneously catabolizes five major components of corn stover lignocellulose: Glucose, xylose, arabinose, p-coumaric acid, and acetic acid

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References (47)

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