The oleaginous yeast Rhodosporidium toruloides engineered for biomass hydrolysate-derived (E)-α-bisabolene production

Paul A. Adamczyk; Hee Jin Hwang; Ta-Hsuan Chang; Yuqian Gao; Edward E.K. Baidoo; Joonhoon Kim; Bobbie-Jo M. Webb-Robertson; Javier E. Flores; Kirch Czarina Quijano; Meagan C. Burnet; Nathalie Munoz; Eric Sundstrom; John M. Gladden; Di Liu

doi:10.1016/j.ymben.2025.02.014

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The oleaginous yeast Rhodosporidium toruloides engineered for biomass hydrolysate-derived (E)-α-bisabolene production

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The oleaginous yeast Rhodosporidium toruloides engineered for biomass hydrolysate-derived (E)-α-bisabolene production

Paul A. Adamczyk, Hee Jin Hwang, Ta-Hsuan Chang, Yuqian Gao, Edward E.K. Baidoo, Joonhoon Kim, Bobbie-Jo M. Webb-Robertson, Javier E. Flores, Kirch Czarina Quijano, Meagan C. Burnet, …

Metabolic engineering, Vol.90, pp.92-105

07/01/2025

DOI: 10.1016/j.ymben.2025.02.014

PMID: 40044027

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Abstract

The oleaginous yeast Rhodosporidiumtoruloides has been exploited for many bioproducts, including several terpenes, owing to its oleaginous nature and biomass inhibitor tolerance. Here, we built upon previous (E)-α-bisabolene work by iteratively stacking the complete mevalonate pathway from Saccharomyces cerevisiae onto a multicopy bisabolene synthase parent strain. Metabolomics and proteomics verified heterologous pathway expression and identified metabolic bottlenecks at three intermediate steps, with candidate feedback-resistant mevalonate kinases screening improving titers 15%. Subtle differences in codon optimization, and preliminary attenuation of competing flux toward lipids resulted in 6-fold, 7-fold higher titers relative to controls, respectively. Media optimization led to modest improvements, with zinc identified as the most promising at 10% titer improvement. Ultimately, high-performance strains were cultivated with corn-stover biomass hydrolysate in microtiter plates at 300 g/L total sugar, achieving 20.8 g/L bisabolene, the highest reported titer in the literature. A 2 L glucose minimal medium bioreactor achieved 19.3 g/L bisabolene and a literature-high productivity of 0.11 g/L/h. •The entire Saccharomyces cerevisiae mevalonate pathway was overexpressed.•Three pathway bottlenecks were identified via multi-omic analysis.•Literature-high titer of 20.8 g/L bisabolene was obtained from 300 g/L total sugar corn stover hydrolysate.•Highest productivity in literature of 0.11 g/L/h from glucose was achieved.

Bisabolene

Lignocellulosic hydrolysate

Mevalonate pathway

Omics

Rhodosporidium toruloides

Rhodotorula

Details

Title: Subtitle: The oleaginous yeast Rhodosporidium toruloides engineered for biomass hydrolysate-derived (E)-α-bisabolene production
Creators: Paul A. Adamczyk - Sandia National Laboratories California
Hee Jin Hwang - Sandia National Laboratories California
Ta-Hsuan Chang - Lawrence Berkeley National Laboratory
Yuqian Gao - Pacific Northwest National Laboratory
Edward E.K. Baidoo - Joint BioEnergy Institute
Joonhoon Kim - Pacific Northwest National Laboratory
Bobbie-Jo M. Webb-Robertson - Agile Biofoundry, Emeryville, CA, USA
Javier E. Flores - Pacific Northwest National Laboratory
Kirch Czarina Quijano - Lawrence Berkeley National Laboratory
Meagan C. Burnet - Pacific Northwest National Laboratory
Nathalie Munoz - Pacific Northwest National Laboratory
Eric Sundstrom - Lawrence Berkeley National Laboratory
John M. Gladden - Joint BioEnergy Institute
Di Liu - Sandia National Laboratories California
Resource Type: Journal article
Publication Details: Metabolic engineering, Vol.90, pp.92-105
DOI: 10.1016/j.ymben.2025.02.014
PMID: 40044027
NLM abbreviation: Metab Eng
ISSN: 1096-7176
eISSN: 1096-7184
Publisher: Elsevier Inc
Number of pages: 14
Language: English
Date published: 07/01/2025
Academic Unit: Biostatistics
Record Identifier: 9985113182002771

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