Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides
Abstract
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 formore »
- Publication Date:
- Research Org.:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1618899
- Alternate Identifier(s):
- OSTI ID: 1600027; OSTI ID: 1604735
- Report Number(s):
- PNNL-SA-150034
Journal ID: ISSN 1475-2859; 24; PII: 1293
- Grant/Contract Number:
- AC02-05CH11231; AC05-76RL01830
- Resource Type:
- Published Article
- Journal Name:
- Microbial Cell Factories
- Additional Journal Information:
- Journal Name: Microbial Cell Factories Journal Volume: 19 Journal Issue: 1; Journal ID: ISSN 1475-2859
- Publisher:
- Springer Science + Business Media
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Rhodotorula; mevalonate pathway; diterpene; geranylgeranyl pyrophosphate synthase; mutant farnesyl pyrophosphate synthase; metabolic engineering
Citation Formats
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., and Gladden, John M. Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides. United Kingdom: N. p., 2020.
Web. doi:10.1186/s12934-020-1293-8.
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. Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides. United Kingdom. https://doi.org/10.1186/s12934-020-1293-8
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., and Gladden, John M. Wed .
"Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides". United Kingdom. https://doi.org/10.1186/s12934-020-1293-8.
@article{osti_1618899,
title = {Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides},
author = {Geiselman, Gina M. and Zhuang, Xun and Kirby, James and Tran-Gyamfi, Mary B. and Prahl, Jan-Philip and Sundstrom, Eric R. and Gao, Yuqian and Munoz Munoz, Nathalie and Nicora, Carrie D. and Clay, Derek M. and Papa, Gabriella and Burnum-Johnson, Kristin E. and Magnuson, Jon K. and Tanjore, Deepti and Skerker, Jeffrey M. and Gladden, John M.},
abstractNote = {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.},
doi = {10.1186/s12934-020-1293-8},
journal = {Microbial Cell Factories},
number = 1,
volume = 19,
place = {United Kingdom},
year = {2020},
month = {2}
}
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1186/s12934-020-1293-8
https://doi.org/10.1186/s12934-020-1293-8
Other availability
Search WorldCat to find libraries that may hold this journal
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.
Works referenced in this record:
Cytotoxicity and Chemotherapeutic Potential of Natural Rosin Abietane Diterpenoids and their Synthetic Derivatives
journal, February 2019
- Faustino, Célia; Neto, Íris; Fonte, Pedro
- Current Pharmaceutical Design, Vol. 24, Issue 36
Biosynthesis and function of terpenoid defense compounds in maize (Zea mays)
journal, September 2018
- Block, Anna K.; Vaughan, Martha M.; Schmelz, Eric A.
- Planta, Vol. 249, Issue 1
BIOCHEMISTRY: Creating Isoprenoid Diversity
journal, September 1997
- Sacchettini, J. C.
- Science, Vol. 277, Issue 5333
Farnesyl Diphosphate Synthase. A Paradigm for Understanding Structure and Function Relationships in E-polyprenyl Diphosphate Synthases
journal, February 2006
- Poulter, C. Dale
- Phytochemistry Reviews, Vol. 5, Issue 1
Natural Cytotoxic Diterpenoids, a Potential Source of Drug Leads for Melanoma Therapy
journal, February 2019
- Prieto, Jose M.; Silveira, Dâmaris
- Current Pharmaceutical Design, Vol. 24, Issue 36
The Phorbol Ester Fraction from Jatropha curcas Seed Oil: Potential and Limits for Crop Protection against Insect Pests
journal, November 2012
- Ratnadass, Alain; Wink, Michael
- International Journal of Molecular Sciences, Vol. 13, Issue 12
The Remarkable Structural Diversity Achieved in ent-Kaurane Diterpenes by Fungal Biotransformations
journal, February 2014
- Takahashi, Jacqueline; Gomes, Dhionne; Lyra, Fernanda
- Molecules, Vol. 19, Issue 2
Using Data Independent Acquisition (DIA) to Model High-responding Peptides for Targeted Proteomics Experiments
journal, June 2015
- Searle, Brian C.; Egertson, Jarrett D.; Bollinger, James G.
- Molecular & Cellular Proteomics, Vol. 14, Issue 9
Rhodosporidium toruloides: a new platform organism for conversion of lignocellulose into terpene biofuels and bioproducts
journal, October 2017
- Yaegashi, Junko; Kirby, James; Ito, Masakazu
- Biotechnology for Biofuels, Vol. 10, Issue 1
Current advances in gibberellic acid (GA3) production, patented technologies and potential applications
journal, August 2018
- Camara, Marcela C.; Vandenberghe, Luciana P. S.; Rodrigues, Cristine
- Planta, Vol. 248, Issue 5
Sustainable bioproduction of the blue pigment indigoidine: Expanding the range of heterologous products in R. toruloides to include non-ribosomal peptides
journal, January 2019
- Wehrs, Maren; Gladden, John M.; Liu, Yuzhong
- Green Chemistry, Vol. 21, Issue 12
RNA interference in the oleaginous yeast Rhodosporidium toruloides
journal, April 2019
- Liu, Xiangjian; Zhang, Yue; Liu, Hongdi
- FEMS Yeast Research, Vol. 19, Issue 3
Gibberellin Metabolism and its Regulation
journal, June 2008
- Yamaguchi, Shinjiro
- Annual Review of Plant Biology, Vol. 59, Issue 1
Gibberellin biosynthesis and metabolism: A convergent route for plants, fungi and bacteria
journal, March 2018
- Salazar-Cerezo, Sonia; Martínez-Montiel, Nancy; García-Sánchez, Jenny
- Microbiological Research, Vol. 208
Engineering Aspergillus nidulans for heterologous ent-kaurene and gamma-terpinene production
journal, April 2016
- Bromann, Kirsi; Toivari, Mervi; Viljanen, Kaarina
- Applied Microbiology and Biotechnology, Vol. 100, Issue 14
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
journal, January 2019
- Xu, Yanqun; Charles, Marie Thérèse; Luo, Zisheng
- Plant, Cell & Environment, Vol. 42, Issue 3
Reprogramming the Chemodiversity of Terpenoid Cyclization by Remolding the Active Site Contour of epi -Isozizaene Synthase
journal, February 2014
- Li, Ruiqiong; Chou, Wayne K. W.; Himmelberger, Julie A.
- Biochemistry, Vol. 53, Issue 7
Concomitant Production of Lipids and Carotenoids in Rhodosporidium toruloides under Osmotic Stress Using Response Surface Methodology
journal, October 2016
- Singh, Gunjan; Jawed, Arshad; Paul, Debarati
- Frontiers in Microbiology, Vol. 7
Skyline: an open source document editor for creating and analyzing targeted proteomics experiments
journal, February 2010
- MacLean, Brendan; Tomazela, Daniela M.; Shulman, Nicholas
- Bioinformatics, Vol. 26, Issue 7
MPLEx: a Robust and Universal Protocol for Single-Sample Integrative Proteomic, Metabolomic, and Lipidomic Analyses
journal, May 2016
- Nakayasu, Ernesto S.; Nicora, Carrie D.; Sims, Amy C.
- mSystems, Vol. 1, Issue 3
Conversion of depolymerized sugars and aromatics from engineered feedstocks by two oleaginous red yeasts
journal, August 2019
- Rodriguez, Alberto; Ersig, Nadine; Geiselman, Gina M.
- Bioresource Technology, Vol. 286
Engineering Rhodosporidium toruloides for increased lipid production : Engineering
journal, November 2015
- Zhang, Shuyan; Skerker, Jeffrey M.; Rutter, Charles D.
- Biotechnology and Bioengineering, Vol. 113, Issue 5
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
journal, January 2016
- Chen, Xiaowen; Kuhn, Erik; Jennings, Edward W.
- Energy & Environmental Science, Vol. 9, Issue 4
Cloning of a Full-length cDNA Encoding ent -Kaurene Synthase from Gibberella fujikuroi : Functional Analysis of a Bifunctional Diterpene Cyclase
journal, January 2000
- Toyomasu, Tomonobu; Kawaide, Hiroshi; Ishizaki, Atsuko
- Bioscience, Biotechnology, and Biochemistry, Vol. 64, Issue 3
Combining metabolic and protein engineering of a terpenoid biosynthetic pathway for overproduction and selectivity control
journal, July 2010
- Leonard, E.; Ajikumar, P. K.; Thayer, K.
- Proceedings of the National Academy of Sciences, Vol. 107, Issue 31, p. 13654-13659
Regulation of product chain length by isoprenyl diphosphate synthases
journal, December 1996
- Tarshis, L. C.; Proteau, P. J.; Kellogg, B. A.
- Proceedings of the National Academy of Sciences, Vol. 93, Issue 26
CHEMISTRY: Roots of Biosynthetic Diversity
journal, April 2007
- Christianson, D. W.
- Science, Vol. 316, Issue 5821
Diterpenoid biopolymers: New directions for renewable materials engineering
journal, September 2010
- Hillwig, Matthew L.; Mann, Francis M.; Peters, Reuben J.
- Biopolymers, Vol. 95, Issue 2
Diterpenes as lead molecules against neglected tropical diseases: Diterpenes Acting Against Neglected Tropical Diseases
journal, November 2016
- Barros de Alencar, Marcus Vinícius Oliveira; de Castro e. Sousa, João Marcelo; Rolim, Hercília Maria Lins
- Phytotherapy Research, Vol. 31, Issue 2
Design, implementation and practice of JBEI-ICE: an open source biological part registry platform and tools
journal, June 2012
- Ham, T. S.; Dmytriv, Z.; Plahar, H.
- Nucleic Acids Research, Vol. 40, Issue 18
Monoterpene production by the carotenogenic yeast Rhodosporidium toruloides
journal, March 2019
- Zhuang, Xun; Kilian, Oliver; Monroe, Eric
- Microbial Cell Factories, Vol. 18, Issue 1
CONSeQuence: Prediction of Reference Peptides for Absolute Quantitative Proteomics Using Consensus Machine Learning Approaches
journal, August 2011
- Eyers, Claire E.; Lawless, Craig; Wedge, David C.
- Molecular & Cellular Proteomics, Vol. 10, Issue 11
High-density cultivation of oleaginous yeast Rhodosporidium toruloides Y4 in fed-batch culture
journal, August 2007
- Li, Yonghong; Zhao, Zongbao (Kent); Bai, Fengwu
- Enzyme and Microbial Technology, Vol. 41, Issue 3
Demonstrating a separation-free process coupling ionic liquid pretreatment, saccharification, and fermentation with Rhodosporidium toruloides to produce advanced biofuels
journal, January 2018
- Sundstrom, Eric; Yaegashi, Junko; Yan, Jipeng
- Green Chemistry, Vol. 20, Issue 12
A highly efficient dilute alkali deacetylation and mechanical (disc) refining process for the conversion of renewable biomass to lower cost sugars
journal, January 2014
- Chen, Xiaowen; Shekiro, Joseph; Pschorn, Thomas
- Biotechnology for Biofuels, Vol. 7, Issue 1
Diterpene synthases facilitating production of the kaurane skeleton of eriocalyxin B in the medicinal plant Isodon eriocalyx
journal, February 2019
- Du, Gang; Gong, Hai-Yan; Feng, Ke-Na
- Phytochemistry, Vol. 158
Effects of biomass hydrolysis by-products on oleaginous yeast Rhodosporidium toruloides
journal, October 2009
- Hu, Cuimin; Zhao, Xin; Zhao, Jin
- Bioresource Technology, Vol. 100, Issue 20
The prenyl transfer reaction. Enzymic and mechanistic studies of the 1'-4 coupling reaction in the terpene biosynthetic pathway
journal, August 1978
- Poulter, C. Dale; Rilling, Hans C.
- Accounts of Chemical Research, Vol. 11, Issue 8
Metabolic engineering of the Stevia rebaudiana ent-kaurene biosynthetic pathway in recombinant Escherichia coli
journal, November 2015
- Kong, Min Kyung; Kang, Hyun-Jun; Kim, Jin Ho
- Journal of Biotechnology, Vol. 214