Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides

Geiselman, Gina M.; Zhuang, Xun; Kirby, James; Tran-Gyamfi, Mary B.; 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 · Wed Feb 05 00:00:00 EST 2020 · Microbial Cell Factories

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

Geiselman, Gina M.; Zhuang, Xun; Kirby, James; Tran-Gyamfi, Mary B.; 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.;

BACKGROUND: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. RESULTS: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. CONCLUSION: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:: AC02-05CH11231; AC05-76RL01830

OSTI ID:: 1618899

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

Report Number(s):: PNNL-SA-150034; 24; PII: 1293

Journal Information:: Microbial Cell Factories, Journal Name: Microbial Cell Factories Vol. 19 Journal Issue: 1; ISSN 1475-2859

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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Cited by: 19 works

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References (36)

Cytotoxicity and Chemotherapeutic Potential of Natural Rosin Abietane Diterpenoids and their Synthetic Derivatives Faustino, Célia; Neto, Íris; Fonte, Pedro Current Pharmaceutical Design, Vol. 24, Issue 36 https://doi.org/10.2174/1381612825666190112162817	journal	February 2019
Biosynthesis and function of terpenoid defense compounds in maize (Zea mays) Block, Anna K.; Vaughan, Martha M.; Schmelz, Eric A. Planta, Vol. 249, Issue 1 https://doi.org/10.1007/s00425-018-2999-2	journal	September 2018
BIOCHEMISTRY: Creating Isoprenoid Diversity Sacchettini, J. C. Science, Vol. 277, Issue 5333 https://doi.org/10.1126/science.277.5333.1788	journal	September 1997
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
Natural Cytotoxic Diterpenoids, a Potential Source of Drug Leads for Melanoma Therapy Prieto, Jose M.; Silveira, Dâmaris Current Pharmaceutical Design, Vol. 24, Issue 36 https://doi.org/10.2174/1381612825666190111143648	journal	February 2019
The Phorbol Ester Fraction from Jatropha curcas Seed Oil: Potential and Limits for Crop Protection against Insect Pests Ratnadass, Alain; Wink, Michael International Journal of Molecular Sciences, Vol. 13, Issue 12 https://doi.org/10.3390/ijms131216157	journal	November 2012
The Remarkable Structural Diversity Achieved in ent-Kaurane Diterpenes by Fungal Biotransformations Takahashi, Jacqueline; Gomes, Dhionne; Lyra, Fernanda Molecules, Vol. 19, Issue 2 https://doi.org/10.3390/molecules19021856	journal	February 2014
Rhodosporidium toruloides: a new platform organism for conversion of lignocellulose into terpene biofuels and bioproducts Yaegashi, Junko; Kirby, James; Ito, Masakazu Biotechnology for Biofuels, Vol. 10, Issue 1 https://doi.org/10.1186/s13068-017-0927-5	journal	October 2017
Current advances in gibberellic acid (GA3) production, patented technologies and potential applications Camara, Marcela C.; Vandenberghe, Luciana P. S.; Rodrigues, Cristine Planta, Vol. 248, Issue 5 https://doi.org/10.1007/s00425-018-2959-x	journal	August 2018
RNA interference in the oleaginous yeast Rhodosporidium toruloides Liu, Xiangjian; Zhang, Yue; Liu, Hongdi FEMS Yeast Research, Vol. 19, Issue 3 https://doi.org/10.1093/femsyr/foz031	journal	April 2019
Gibberellin Metabolism and its Regulation Yamaguchi, Shinjiro Annual Review of Plant Biology, Vol. 59, Issue 1 https://doi.org/10.1146/annurev.arplant.59.032607.092804	journal	June 2008
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
Engineering Aspergillus nidulans for heterologous ent-kaurene and gamma-terpinene production Bromann, Kirsi; Toivari, Mervi; Viljanen, Kaarina Applied Microbiology and Biotechnology, Vol. 100, Issue 14 https://doi.org/10.1007/s00253-016-7517-5	journal	April 2016
Ultraviolet-C priming of strawberry leaves against subsequent Mycosphaerella fragariae infection involves the action of reactive oxygen species, plant hormones, and terpenes: Priming of strawberry plants by ultraviolet light Xu, Yanqun; Charles, Marie Thérèse; Luo, Zisheng Plant, Cell & Environment, Vol. 42, Issue 3 https://doi.org/10.1111/pce.13491	journal	January 2019
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
Concomitant Production of Lipids and Carotenoids in Rhodosporidium toruloides under Osmotic Stress Using Response Surface Methodology Singh, Gunjan; Jawed, Arshad; Paul, Debarati Frontiers in Microbiology, Vol. 7 https://doi.org/10.3389/fmicb.2016.01686	journal	October 2016
Skyline: an open source document editor for creating and analyzing targeted proteomics experiments MacLean, Brendan; Tomazela, Daniela M.; Shulman, Nicholas Bioinformatics, Vol. 26, Issue 7 https://doi.org/10.1093/bioinformatics/btq054	journal	February 2010
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
Engineering Rhodosporidium toruloides for increased lipid production : Engineering Zhang, Shuyan; Skerker, Jeffrey M.; Rutter, Charles D. Biotechnology and Bioengineering, Vol. 113, Issue 5 https://doi.org/10.1002/bit.25864	journal	November 2015
DMR (deacetylation and mechanical refining) processing of corn stover achieves high monomeric sugar concentrations (230 g L ⁻¹ ) during enzymatic hydrolysis and high ethanol concentrations (>10% v/v) during fermentation without hydrolysate purification or concentration Chen, Xiaowen; Kuhn, Erik; Jennings, Edward W. Energy & Environmental Science, Vol. 9, Issue 4 https://doi.org/10.1039/C5EE03718B	journal	January 2016
Cloning of a Full-length cDNA Encoding ent -Kaurene Synthase from Gibberella fujikuroi : Functional Analysis of a Bifunctional Diterpene Cyclase Toyomasu, Tomonobu; Kawaide, Hiroshi; Ishizaki, Atsuko Bioscience, Biotechnology, and Biochemistry, Vol. 64, Issue 3 https://doi.org/10.1271/bbb.64.660	journal	January 2000
Combining metabolic and protein engineering of a terpenoid biosynthetic pathway for overproduction and selectivity control Leonard, E.; Ajikumar, P. K.; Thayer, K. Proceedings of the National Academy of Sciences, Vol. 107, Issue 31, p. 13654-13659 https://doi.org/10.1073/pnas.1006138107	journal	July 2010
Regulation of product chain length by isoprenyl diphosphate synthases Tarshis, L. C.; Proteau, P. J.; Kellogg, B. A. Proceedings of the National Academy of Sciences, Vol. 93, Issue 26 https://doi.org/10.1073/pnas.93.26.15018	journal	December 1996
CHEMISTRY: Roots of Biosynthetic Diversity Christianson, D. W. Science, Vol. 316, Issue 5821 https://doi.org/10.1126/science.1141630	journal	April 2007
Diterpenoid biopolymers: New directions for renewable materials engineering Hillwig, Matthew L.; Mann, Francis M.; Peters, Reuben J. Biopolymers, Vol. 95, Issue 2 https://doi.org/10.1002/bip.21538	journal	September 2010
Diterpenes as lead molecules against neglected tropical diseases: Diterpenes Acting Against Neglected Tropical Diseases Barros de Alencar, Marcus Vinícius Oliveira; de Castro e. Sousa, João Marcelo; Rolim, Hercília Maria Lins Phytotherapy Research, Vol. 31, Issue 2 https://doi.org/10.1002/ptr.5749	journal	November 2016
Design, implementation and practice of JBEI-ICE: an open source biological part registry platform and tools Ham, T. S.; Dmytriv, Z.; Plahar, H. Nucleic Acids Research, Vol. 40, Issue 18 https://doi.org/10.1093/nar/gks531	journal	June 2012
Monoterpene production by the carotenogenic yeast Rhodosporidium toruloides Zhuang, Xun; Kilian, Oliver; Monroe, Eric Microbial Cell Factories, Vol. 18, Issue 1 https://doi.org/10.1186/s12934-019-1099-8	journal	March 2019
CONSeQuence: Prediction of Reference Peptides for Absolute Quantitative Proteomics Using Consensus Machine Learning Approaches Eyers, Claire E.; Lawless, Craig; Wedge, David C. Molecular & Cellular Proteomics, Vol. 10, Issue 11 https://doi.org/10.1074/mcp.M110.003384	journal	August 2011
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
Demonstrating a separation-free process coupling ionic liquid pretreatment, saccharification, and fermentation with Rhodosporidium toruloides to produce advanced biofuels Sundstrom, Eric; Yaegashi, Junko; Yan, Jipeng Green Chemistry, Vol. 20, Issue 12 https://doi.org/10.1039/C8GC00518D	journal	January 2018
A highly efficient dilute alkali deacetylation and mechanical (disc) refining process for the conversion of renewable biomass to lower cost sugars Chen, Xiaowen; Shekiro, Joseph; Pschorn, Thomas Biotechnology for Biofuels, Vol. 7, Issue 1 https://doi.org/10.1186/1754-6834-7-98	journal	January 2014
Diterpene synthases facilitating production of the kaurane skeleton of eriocalyxin B in the medicinal plant Isodon eriocalyx Du, Gang; Gong, Hai-Yan; Feng, Ke-Na Phytochemistry, Vol. 158 https://doi.org/10.1016/j.phytochem.2018.11.015	journal	February 2019
Effects of biomass hydrolysis by-products on oleaginous yeast Rhodosporidium toruloides Hu, Cuimin; Zhao, Xin; Zhao, Jin Bioresource Technology, Vol. 100, Issue 20 https://doi.org/10.1016/j.biortech.2009.04.041	journal	October 2009
The prenyl transfer reaction. Enzymic and mechanistic studies of the 1'-4 coupling reaction in the terpene biosynthetic pathway Poulter, C. Dale; Rilling, Hans C. Accounts of Chemical Research, Vol. 11, Issue 8 https://doi.org/10.1021/ar50128a004	journal	August 1978
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

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59 BASIC BIOLOGICAL SCIENCES
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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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