The in vivo synthesis of plant sesquiterpenes by Escherichia coli
Abstract
Three plant genes encoding (+)‐δ‐cadinene, 5‐epi‐aristolochene, and vetispiradiene cyclases were expressed in Escherichia coli to evaluate the potential of this bacterium to synthesize sesquiterpenes in vivo. Various growth temperatures, carbon sources, and host strains were examined to optimize terpene production. The highest levels of sesquiterpene production occurred when the enzymes were expressed in strain DH5α from the trc promoter (Ptrc) of the high‐copy plasmidpTrc99A in M9 medium supplemented with 0.2% (v/v) glycerol at 30°C for 5‐epi‐aristolochene and vetispiradiene and 37°C for (+)‐δ‐cadinene. The highest concentrations of sesquiterpenes observed were 10.3 μg of (+)‐δ‐cadinene, 0.24 μg of 5‐epi‐aristolochene (measured as (+)‐δ‐cadinene equivalents), and 6.4 μg of vetispiradiene (measured as (+)‐δ‐cadinene equivalents) per liter of culture. These sesquiterpene production levels are >500‐fold lower than carotenoid production, both of which are synthesized from endogenous trans‐farnesyl diphosphate (FDP) in E. coli. Based on these results, we conclude that the limiting factor for sesquiterpene synthesis in E. coli is the poor expression of the cyclase enzyme and not supply of the FDP precursor. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 497–503, 2001.
Citing Literature
Number of times cited: 54
- Ozkan Fidan and Jixun Zhan, Reconstitution of Medicinally Important Plant Natural Products in Microorganisms, Molecular Pharming, (383-415), (2018).
- Xiaoping Tang, Melodi Demiray, Thomas Wirth and Rudolf K. Allemann, Concise synthesis of artemisinin from a farnesyl diphosphate analogue, Bioorganic & Medicinal Chemistry, 10.1016/j.bmc.2017.03.068, (2017).
- Xixian Chen, Congqiang Zhang, Ruiyang Zou, Gregory Stephanopoulos and Heng-Phon Too, In Vitro Metabolic Engineering of Amorpha-4,11-diene Biosynthesis at Enhanced Rate and Specific Yield of Production , ACS Synthetic Biology, 10.1021/acssynbio.6b00377, 6, 9, (1691-1700), (2017).
- Amit Rai, Kazuki Saito and Mami Yamazaki, Integrated omics analysis of specialized metabolism in medicinal plants, The Plant Journal, 90, 4, (764-787), (2017).
- S. Eric Nybo, Jacqueline Saunders and Sean P. McCormick, Metabolic engineering of Escherichia coli for production of valerenadiene, Journal of Biotechnology, 262, (60), (2017).
- Stefan Tippmann, Gionata Scalcinati, Verena Siewers and Jens Nielsen, Production of farnesene and santalene by Saccharomyces cerevisiae using fed‐batch cultivations with RQ‐controlled feed, Biotechnology and Bioengineering, 113, 1, (72-81), (2015).
- Aamir Rasool, Genlin Zhang, Zhe Li and Chun Li, Engineering of the terpenoid pathway in Saccharomyces cerevisiae co-overproduces squalene and the non-terpenoid compound oleic acid, Chemical Engineering Science, 10.1016/j.ces.2016.06.004, 152, (457-467), (2016).
- Aamir Rasool, Muhammad Saad Ahmed and Chun Li, Overproduction of squalene synergistically downregulates ethanol production in Saccharomyces cerevisiae, Chemical Engineering Science, 152, (370), (2016).
- Han Xiao and Jian-Jiang Zhong, Production of Useful Terpenoids by Higher-Fungus Cell Factory and Synthetic Biology Approaches, Trends in Biotechnology, 10.1016/j.tibtech.2015.12.007, 34, 3, (242-255), (2016).
- Zou Huibin, Hui Liu, Elhussiny Aboulnaga, Huizhou Liu, Tao Cheng and Mo Xian, Microbial Production of Isoprene: Opportunities and Challenges, Industrial Biotechnology, (473-504), (2016).
- Jianming Yang, Zhengfeng Li, Lizhong Guo, Juan Du and Hyeun-Jong Bae, Biosynthesis of β-caryophyllene, a novel terpene-based high-density biofuel precursor, using engineered Escherichia coli, Renewable Energy, 10.1016/j.renene.2016.06.061, 99, (216-223), (2016).
- Julia Gallego-Jara, Teresa de Diego, Álvaro del Real, Ana Écija-Conesa, Arturo Manjón and Manuel Cánovas, Lycopene overproduction and in situ extraction in organic-aqueous culture systems using a metabolically engineered Escherichia coli, AMB Express, 5, 1, (2015).
- Murillo L. Bution, Gustavo Molina, Meissa R.E. Abrahão and Gláucia M. Pastore, Genetic and metabolic engineering of microorganisms for the development of new flavor compounds from terpenic substrates, Critical Reviews in Biotechnology, 35, 3, (313), (2015).
- Xinxiao Sun, Xiaolin Shen, Rachit Jain, Yuheng Lin, Jian Wang, Jing Sun, Jia Wang, Yajun Yan and Qipeng Yuan, Synthesis of chemicals by metabolic engineering of microbes, Chemical Society Reviews, 10.1039/C5CS00159E, 44, 11, (3760-3785), (2015).
- Pooja Jambunathan and Kechun Zhang, Combining biological and chemical approaches for green synthesis of chemicals, Current Opinion in Chemical Engineering, 10, (35), (2015).
- Maiko Furubayashi, Mayu Ikezumi, Jun Kajiwara, Miki Iwasaki, Akira Fujii, Ling Li, Kyoichi Saito, Daisuke Umeno and Danilo Roccatano, A High-Throughput Colorimetric Screening Assay for Terpene Synthase Activity Based on Substrate Consumption, PLoS ONE, 9, 3, (e93317), (2014).
- Adelene A. Song, Janna Ong Abdullah, Mohd. P. Abdullah, Norazizah Shafee, Roohaida Othman, Normah Mohd. Noor and Raha A. Rahim, Engineering the lactococcal mevalonate pathway for increased sesquiterpene production, FEMS Microbiology Letters, 355, 2, (177-184), (2014).
- Shiro Suzuki, Takao Koeduka, Akifumi Sugiyama, Kazufumi Yazaki and Toshiaki Umezawa, Microbial production of plant specialized metabolites, Plant Biotechnology, 10.5511/plantbiotechnology.14.1003a, 31, 5, (465-482), (2014).
- Lishan Zhao, Wei-chen Chang, Youli Xiao, Hung-wen Liu and Pinghua Liu, Methylerythritol Phosphate Pathway of Isoprenoid Biosynthesis, Annual Review of Biochemistry, 10.1146/annurev-biochem-052010-100934, 82, 1, (497-530), (2013).
- Bilge Özaydın, Helcio Burd, Taek Soon Lee and Jay D. Keasling, Carotenoid-based phenotypic screen of the yeast deletion collection reveals new genes with roles in isoprenoid production, Metabolic Engineering, 10.1016/j.ymben.2012.07.010, 15, (174-183), (2013).
- Robert H Dahl, Fuzhong Zhang, Jorge Alonso-Gutierrez, Edward Baidoo, Tanveer S Batth, Alyssa M Redding-Johanson, Christopher J Petzold, Aindrila Mukhopadhyay, Taek Soon Lee, Paul D Adams and Jay D Keasling, Engineering dynamic pathway regulation using stress-response promoters, Nature Biotechnology, 31, 11, (1039), (2013).
- Jay D. Keasling, Synthetic biology and the development of tools for metabolic engineering, Metabolic Engineering, 10.1016/j.ymben.2012.01.004, 14, 3, (189-195), (2012).
- Sotirios C. Kampranis and Antonios M. Makris, DEVELOPING A YEAST CELL FACTORY FOR THE PRODUCTION OF TERPENOIDS, Computational and Structural Biotechnology Journal, 3, 4, (e201210006), (2012).
- Adelene Ai-Lian Song, Janna Ong Abdullah, Mohd. Puad Abdullah, Norazizah Shafee, Roohaida Othman, Ee-Fun Tan, Normah Mohd. Noor, Abdul Rahim Raha and Paul Jaak Janssen, Overexpressing 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase (HMGR) in the Lactococcal Mevalonate Pathway for Heterologous Plant Sesquiterpene Production, PLoS ONE, 7, 12, (e52444), (2012).
- Karine Berthelot, Yannick Estevez, Alain Deffieux and Frédéric Peruch, Isopentenyl diphosphate isomerase: A checkpoint to isoprenoid biosynthesis, Biochimie, 10.1016/j.biochi.2012.03.021, 94, 8, (1621-1634), (2012).
- Haoran Zhang, Brett A. Boghigian, John Armando and Blaine A. Pfeifer, Methods and options for the heterologous production of complex natural products, Nat. Prod. Rep., 10.1039/C0NP00037J, 28, 1, (125-151), (2011).
- Claudia Schmidt‐Dannert and Alexander Pisarchik, Biosynthetic Pathway Engineering Strategies, Protein Engineering Handbook, (849-876), (2011).
- Chonglong Wang, Sang-Hwal Yoon, Hui-Jeong Jang, Young-Ryun Chung, Jae-Yean Kim, Eui-Sung Choi and Seon-Won Kim, Metabolic engineering of Escherichia coli for α-farnesene production, Metabolic Engineering, 10.1016/j.ymben.2011.08.001, 13, 6, (648-655), (2011).
- Yeong-Su Kim, Jae-Hee Lee, Nam-Hee Kim, Soo-Jin Yeom, Seon-Won Kim and Deok-Kun Oh, Increase of lycopene production by supplementing auxiliary carbon sources in metabolically engineered Escherichia coli, Applied Microbiology and Biotechnology, 90, 2, (489), (2011).
- Chonglong Wang, Sang‐Hwal Yoon, Asad Ali Shah, Young‐Ryun Chung, Jae‐Yean Kim, Eui‐Sung Choi, Jay D. Keasling and Seon‐Won Kim, Farnesol production from Escherichia coli by harnessing the exogenous mevalonate pathway, Biotechnology and Bioengineering, 107, 3, (421-429), (2010).
- Sang-Hwal Yoon, Sook-Hee Lee, Amitabha Das, Hee-Kyoung Ryu, Hee-Jeong Jang, Jae-Yean Kim, Deok-Kun Oh, Jay D. Keasling and Seon-Won Kim, Combinatorial expression of bacterial whole mevalonate pathway for the production of β-carotene in E. coli, Journal of Biotechnology, 10.1016/j.jbiotec.2009.01.008, 140, 3-4, (218-226), (2009).
- Sang Yup Lee, Hyun Uk Kim, Jin Hwan Park, Jong Myung Park and Tae Yong Kim, Metabolic engineering of microorganisms: general strategies and drug production, Drug Discovery Today, 14, 1-2, (78), (2009).
- Collin H. Martin, David R. Nielsen, Kevin V. Solomon and Kristala L. Jones Prather, Synthetic Metabolism: Engineering Biology at the Protein and Pathway Scales, Chemistry & Biology, 10.1016/j.chembiol.2009.01.010, 16, 3, (277-286), (2009).
- Sung-Woo Kim, Jae-Bum Kim, Ji-Myung Ryu, Joon-Ki Jung and Jung-Hoe Kim, High-level production of lycopene in metabolically engineered E. coli, Process Biochemistry, 10.1016/j.procbio.2009.04.018, 44, 8, (899-905), (2009).
- Eric M. Paradise, James Kirby, Rossana Chan and Jay D. Keasling, Redirection of flux through the FPP branch‐point in Saccharomyces cerevisiae by down‐regulating squalene synthase, Biotechnology and Bioengineering, 100, 2, (371-378), (2008).
- J.L. Fortman, Swapnil Chhabra, Aindrila Mukhopadhyay, Howard Chou, Taek Soon Lee, Eric Steen and Jay D. Keasling, Biofuel alternatives to ethanol: pumping the microbial well, Trends in Biotechnology, 10.1016/j.tibtech.2008.03.008, 26, 7, (375-381), (2008).
- David Lubertozzi and Jay D. Keasling, Expression of a synthetic Artemesia annua amorphadiene synthase in Aspergillus nidulans yields altered product distribution, Journal of Industrial Microbiology & Biotechnology, 35, 10, (1191), (2008).
- Mohammad A. Asadollahi, Jérôme Maury, Kasper Møller, Kristian Fog Nielsen, Michel Schalk, Anthony Clark and Jens Nielsen, Production of plant sesquiterpenes in Saccharomyces cerevisiae: Effect of ERG9 repression on sesquiterpene biosynthesis, Biotechnology and Bioengineering, 99, 3, (666-677), (2007).
- Qingping Zeng, Frank Qiu and Ling Yuan, Production of artemisinin by genetically-modified microbes, Biotechnology Letters, 30, 4, (581), (2008).
- Yasuo Yoshikuni, Jeffrey A. Dietrich, Farnaz F. Nowroozi, Patricia C. Babbitt and Jay D. Keasling, Redesigning Enzymes Based on Adaptive Evolution for Optimal Function in Synthetic Metabolic Pathways, Chemistry & Biology, 15, 6, (607), (2008).
- Yasuo Yoshikuni and Jay D Keasling, Pathway engineering by designed divergent evolution, Current Opinion in Chemical Biology, 11, 2, (233), (2007).
- Amitabha Das, Sang-Hwal Yoon, Sook-Hee Lee, Jae-Yean Kim, Deok-Kun Oh and Seon-Won Kim, An update on microbial carotenoid production: application of recent metabolic engineering tools, Applied Microbiology and Biotechnology, 10.1007/s00253-007-1206-3, 77, 3, (505-512), (2007).
- Yoichiro Shiba, Eric M. Paradise, James Kirby, Dae-Kyun Ro and Jay D. Keasling, Engineering of the pyruvate dehydrogenase bypass in Saccharomyces cerevisiae for high-level production of isoprenoids, Metabolic Engineering, 10.1016/j.ymben.2006.10.005, 9, 2, (160-168), (2007).
- Sydnor T. Withers and Jay D. Keasling, Biosynthesis and engineering of isoprenoid small molecules, Applied Microbiology and Biotechnology, 73, 5, (980), (2006).
- Mattijs K. Julsing, Albert Koulman, Herman J. Woerdenbag, Wim J. Quax and Oliver Kayser, Combinatorial biosynthesis of medicinal plant secondary metabolites, Biomolecular Engineering, 10.1016/j.bioeng.2006.08.001, 23, 6, (265-279), (2006).
- Yasuo Yoshikuni, Vincent J.J. Martin, Thomas E. Ferrin and Jay D. Keasling, Engineering Cotton (+)-δ-Cadinene Synthase to an Altered Function: Germacrene D-4-ol Synthase, Chemistry & Biology, 13, 1, (91), (2006).
- Michelle C Y Chang and Jay D Keasling, Production of isoprenoid pharmaceuticals by engineered microbes, Nature Chemical Biology, 10.1038/nchembio836, 2, 12, (674-681), (2006).
- Beth E. Jackson, Elizabeth A. Hart-Wells and Seiichi P. T. Matsuda, Metabolic Engineering to Produce Sesquiterpenes in Yeast, Organic Letters, 5, 10, (1629), (2003).
- Vincent J J Martin, Douglas J Pitera, Sydnor T Withers, Jack D Newman and Jay D Keasling, Engineering a mevalonate pathway in Escherichia coli for production of terpenoids, Nature Biotechnology, 21, 7, (796), (2003).
- Jan B. van Beilen, Wouter A. Duetz, Andreas Schmid and Bernard Witholt, Practical issues in the application of oxygenases, Trends in Biotechnology, 21, 4, (170), (2003).
- Adelene Ai Lian Song, Janna O. Abdullah, Mohd Puad Abdullah, Norazizah Shafee and Raha A. Rahim, Functional Expression of an Orchid Fragrance Gene in Lactococcus lactis, International Journal of Molecular Sciences, 10.3390/ijms13021582, 13, 12, (1582-1597), (2012).
- Aparajita Banerjee, Alyssa L. Preiser, Thomas D. Sharkey and Eugene A. Permyakov, Engineering of Recombinant Poplar Deoxy-D-Xylulose-5-Phosphate Synthase (PtDXS) by Site-Directed Mutagenesis Improves Its Activity, PLOS ONE, 10.1371/journal.pone.0161534, 11, 8, (e0161534), (2016).
- Xixian Chen, Congqiang Zhang, Ruiyang Zou, Kang Zhou, Gregory Stephanopoulos, Heng Phon Too and Vishal Shah, Statistical Experimental Design Guided Optimization of a One-Pot Biphasic Multienzyme Total Synthesis of Amorpha-4,11-diene, PLoS ONE, 10.1371/journal.pone.0079650, 8, 11, (e79650), (2013).
- Chris J. Paddon and Jay D. Keasling, Semi-synthetic artemisinin: a model for the use of synthetic biology in pharmaceutical development, Nature Reviews Microbiology, 10.1038/nrmicro3240, 12, 5, (355-367), (2014)., (2014).
