Engineering Rhodosporidium toruloides for production of 3-hydroxypropionic acid from lignocellulosic hydrolysate

Liu, Di; Hwang, Hee Jin; Otoupal, Peter B.; Geiselman, Gina M.; Kim, Joonhoon; Pomraning, Kyle R.; Kim, Young-Mo; Munoz, Nathalie; Nicora, Carrie D.; Gao, Yuqian; Burnum-Johnson, Kristin E.; Jacobson, Oslo; Coradetti, Samuel T.; Kim, Jinho; Deng, Shuang; Dai, Ziyu; Prahl, Jan-Philip; Tanjore, Deepti; Lee, Taek Soon; Magnuson, Jon K.; Gladden, John M.

doi:10.1016/j.ymben.2023.05.001

Engineering Rhodosporidium toruloides for production of 3-hydroxypropionic acid from lignocellulosic hydrolysate

Journal Article · Tue May 16 00:00:00 EDT 2023 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2023.05.001· OSTI ID:1995959

^[1]; Hwang, Hee Jin ^[1]; Otoupal, Peter B. ^[2]; Geiselman, Gina M. ^[2]; ^[3]; ^[3]; ^[3]; Munoz, Nathalie ^[3]; ^[4]; Gao, Yuqian ^[3]; ^[3]; Jacobson, Oslo ^[5]; Coradetti, Samuel T. ^[1]; Kim, Jinho ^[6]; Deng, Shuang ^[3]; Dai, Ziyu ^[3]; Prahl, Jan-Philip ^[3]; Tanjore, Deepti ^[3]; Lee, Taek Soon ^[6]; ^[7] more »

Sandia National Lab. (SNL-CA), Livermore, CA (United States); USDOE Agile BioFoundry, Emeryville, CA (United States)
Sandia National Lab. (SNL-CA), Livermore, CA (United States); USDOE Agile BioFoundry, Emeryville, CA (United States); Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
USDOE Agile BioFoundry, Emeryville, CA (United States); Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
USDOE Agile BioFoundry, Emeryville, CA (United States) ; Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
USDOE Agile BioFoundry, Emeryville, CA (United States); Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)

Microbial production of valuable bioproducts is a promising route towards green and sustainable manufacturing. The oleaginous yeast, Rhodosporidium toruloides, has emerged as an attractive host for the production of biofuels and bioproducts from lignocellulosic hydrolysates. 3-hydroxypropionic acid (3HP) is an attractive platform molecule that can be used to produce a wide range of commodity chemicals. This study focuses on establishing and optimizing the production of 3HP in R. toruloides. As R. toruloides naturally has a high metabolic flux towards malonyl-CoA, we exploited this pathway to produce 3HP. Upon finding the yeast capable of catabolizing 3HP, we then implemented functional genomics and metabolomic analysis to identify the catabolic pathways. Deletion of a putative malonate semialdehyde dehydrogenase gene encoding an oxidative 3HP pathway was found to significantly reduce 3HP degradation. We further explored monocarboxylate transporters to promote 3HP transport and identified a novel 3HP transporter in Aspergillus pseudoterreus by RNA-seq and proteomics. Combining these engineering efforts with media optimization in a fed-batch fermentation resulted in 45.4 g/L 3HP production. This represents one of the highest 3HP titers reported, and the highest titer produced from lignocellulosic hydrolysate to the best of our knowledge. This work demonstrates microbial production of 3HP in R. toruloides from lignocellulosic hydrolysate at high titers, and it represents a significant step toward enabling industrial production of 3HP in the future.

View Accepted Manuscript (DOE)

Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)

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

Grant/Contract Number:: AC05-76RL01830; AC02-05CH11231; NA0003525

OSTI ID:: 1995959

Report Number(s):: PNNL-SA-185285

Journal Information:: Metabolic Engineering, Journal Name: Metabolic Engineering Vol. 78; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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3-Hydroxypropionic acid
3-Hydroxypropionic acid transporter
59 BASIC BIOLOGICAL SCIENCES
Lignocellulosic hydrolysate
Malonyl-CoA reductase
R. toruloides

Engineering Rhodosporidium toruloides for production of 3-hydroxypropionic acid from lignocellulosic hydrolysate

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