Abstract

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. toruloidesmore » from lignocellulosic hydrolysate at high titers, and it represents a significant step toward enabling industrial production of 3HP in the future.« less

Authors:

Liu, Di  ^[1];

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• Search DOE PAGES for ORCID "0000-0002-7484-5810"

Hwang, Hee Jin ^[1]; Otoupal, Peter B. ^[2]; Geiselman, Gina M. ^[2]; Kim, Joonhoon  ^[3];

• Search DOE PAGES for author "Kim, Joonhoon"
• Search DOE PAGES for ORCID "0000-0002-7425-1828"

Pomraning, Kyle R.  ^[3];

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• Search DOE PAGES for ORCID "0000-0003-2324-2881"

Kim, Young-Mo  ^[3];

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• Search DOE PAGES for ORCID "0000-0002-8972-7593"

Munoz, Nathalie ^[3]; Nicora, Carrie D.  ^[4];

• Search DOE PAGES for author "Nicora, Carrie D."
• Search DOE PAGES for ORCID "0000-0003-2461-9548"

Gao, Yuqian ^[3]; Burnum-Johnson, Kristin E.  ^[3];

• Search DOE PAGES for author "Burnum-Johnson, Kristin E."
• Search DOE PAGES for ORCID "0000-0002-2722-4149"

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]; Magnuson, Jon K.  ^[7]

• Search DOE PAGES for author "Magnuson, Jon K."
• Search DOE PAGES for ORCID "0000-0001-7712-7024"

more »; Gladden, John M.  ^[2]

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• Search DOE PAGES for ORCID "0000-0002-6985-2485"

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+ Show Author Affiliations

1. Sandia National Lab. (SNL-CA), Livermore, CA (United States); USDOE Agile BioFoundry, Emeryville, CA (United States)
2. 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)
3. USDOE Agile BioFoundry, Emeryville, CA (United States); Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
4. Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
5. USDOE Agile BioFoundry, Emeryville, CA (United States) ; Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
6. Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
7. 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)

Publication Date:

Tue May 16 00:00:00 EDT 2023

Research Org.:

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

Sponsoring Org.:

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)

OSTI Identifier:

1995959

Report Number(s):

PNNL-SA-185285
Journal ID: ISSN 1096-7176

Grant/Contract Number:  

AC05-76RL01830; AC02-05CH11231; NA0003525

Resource Type:

Accepted Manuscript

Journal Name:

Metabolic Engineering

Additional Journal Information:

Journal Volume: 78; Journal ID: ISSN 1096-7176

Publisher:

Elsevier

Country of Publication:

United States

Language:

English

Subject:

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