Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides

Geiselman, Gina M.; Zhuang, Xun; Kirby, James; Tran-Gyamfi, Mary; Prahl, Jan-Philip; Sundstrom, Eric R.; Gao, Yuqian; Munoz Munoz, Nathalie; Nicora, Carrie D.; Clay, Derek M.; Papa, Gabriella; Burnum-Johnson, Kristin E.; Magnuson, Jon K.; Tanjore, Deepti; Skerker, Jeffrey M.; Gladden, John M.

doi:10.1186/s12934-020-1293-8

Title: Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides

Journal Article · 05 February 2020 · Microbial Cell Factories

DOI: https://doi.org/10.1186/s12934-020-1293-8 · OSTI ID:1618899

Geiselman, Gina M. ^[1]; Zhuang, Xun ^[1]; Kirby, James ^[2]; Tran-Gyamfi, Mary ^[3]; Prahl, Jan-Philip ^[3]; Sundstrom, Eric R. ^[3]; Gao, Yuqian ^[4]; Munoz Munoz, Nathalie ^[4];

^[4]; Clay, Derek M. ^[3]; Papa, Gabriella ^[5];

^[4];

^[4]; Tanjore, Deepti ^[3]; Skerker, Jeffrey M. ^[3]; Gladden, John M. ^[1]

Sandia National Lab. (SNL-CA), Livermore, CA (United States)
Univ. of California, Berkeley, CA (United States)
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Joint BioEnergy Institute /Sandia National Laboratory

Rhodosporidium toruloides has emerged as a promising host for the production of bioproducts from lignocellulose, in part due to its ability to grow on lignocellulosic feedstocks, tolerate growth inhibitors, and co-utilize sugars and lignin-derived monomers. Ent-kaurene derivatives have a diverse range of potential applications from therapeutics to novel resin-based materials. The Design, Build, Test, and Learn (DBTL) approach was employed to engineer production of the non-native diterpene ent-kaurene in R. toruloides. Following expression of kaurene synthase (KS) in R. toruloides in the first DBTL cycle, a key limitation appeared to be the availability of the diterpene precursor, geranylgeranyl diphosphate (GGPP). Further DBTL cycles were carried out to select an optimal GGPP synthase and to balance its expression with KS, requiring two of the strongest promoters in R. toruloides, ANT (adenine nucleotide translocase) and TEF1 (translational elongation factor 1) to drive expression of the KS from Gibberella fujikuroi and a mutant version of an FPP synthase from Gallus gallus that produces GGPP. Scale-up of cultivation in a 2 L bioreactor using a corn stover hydrolysate resulted in an ent-kaurene titer of 1.4 g/L. This study builds upon previous work demonstrating the potential of R. toruloides as a robust and versatile host for the production of both mono- and sesquiterpenes, and is the first demonstration of the production of a non-native diterpene in this organism.

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Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

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

OSTI ID:: 1618899

Alternate ID(s):: OSTI ID: 1600027; OSTI ID: 1604735

Report Number(s):: PNNL-SA-150034; 1475-2859

Journal Information:: Microbial Cell Factories, Vol. 19

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 24 works

Citation information provided by
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Farnesyl Diphosphate Synthase. A Paradigm for Understanding Structure and Function Relationships in E-polyprenyl Diphosphate Synthases Poulter, C. Dale Phytochemistry Reviews, Vol. 5, Issue 1 https://doi.org/10.1007/s11101-005-4887-1	journal	February 2006
Conversion of depolymerized sugars and aromatics from engineered feedstocks by two oleaginous red yeasts Rodriguez, Alberto; Ersig, Nadine; Geiselman, Gina M. Bioresource Technology, Vol. 286 https://doi.org/10.1016/j.biortech.2019.121365	journal	August 2019
High-density cultivation of oleaginous yeast Rhodosporidium toruloides Y4 in fed-batch culture Li, Yonghong; Zhao, Zongbao (Kent); Bai, Fengwu Enzyme and Microbial Technology, Vol. 41, Issue 3 https://doi.org/10.1016/j.enzmictec.2007.02.008	journal	August 2007
Metabolic engineering of the Stevia rebaudiana ent-kaurene biosynthetic pathway in recombinant Escherichia coli Kong, Min Kyung; Kang, Hyun-Jun; Kim, Jin Ho Journal of Biotechnology, Vol. 214 https://doi.org/10.1016/j.jbiotec.2015.09.016	journal	November 2015
Gibberellin biosynthesis and metabolism: A convergent route for plants, fungi and bacteria Salazar-Cerezo, Sonia; Martínez-Montiel, Nancy; García-Sánchez, Jenny Microbiological Research, Vol. 208 https://doi.org/10.1016/j.micres.2018.01.010	journal	March 2018
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Reprogramming the Chemodiversity of Terpenoid Cyclization by Remolding the Active Site Contour of epi -Isozizaene Synthase Li, Ruiqiong; Chou, Wayne K. W.; Himmelberger, Julie A. Biochemistry, Vol. 53, Issue 7 https://doi.org/10.1021/bi401643u	journal	February 2014
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59 BASIC BIOLOGICAL SCIENCES
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diterpene
geranylgeranyl pyrophosphate synthase
mutant farnesyl pyrophosphate synthase
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Additional file 1 of Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides Geiselman, Gina; Zhuang, Xun; Kirby, James https://doi.org/10.6084/m9.figshare.11814117 The current record is supplemented by this text	text	January 2020
Additional file 1 of Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides Geiselman, Gina; Zhuang, Xun; Kirby, James https://doi.org/10.6084/m9.figshare.11814117.v1 The current record is supplemented by this text	text	January 2020
Additional file 2 of Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides Geiselman, Gina; Zhuang, Xun; Kirby, James https://doi.org/10.6084/m9.figshare.11814120 The current record is supplemented by this text	text	January 2020
Additional file 2 of Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides Geiselman, Gina; Zhuang, Xun; Kirby, James https://doi.org/10.6084/m9.figshare.11814120.v1 The current record is supplemented by this text	text	January 2020
Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides Geiselman, Gina; Zhuang, Xun; Kirby, James https://doi.org/10.6084/m9.figshare.c.4845561 The current record is supplemented by this collection	collection	January 2020
Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides Geiselman, Gina; Zhuang, Xun; Kirby, James https://doi.org/10.6084/m9.figshare.c.4845561.v1 The current record is supplemented by this collection	collection	January 2020

Title: Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides

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