Developing Commercial Production of Semi-Synthetic Artemisinin, and of β-Farnesene, an Isoprenoid Produced by Fermentation of Brazilian Sugar

Benjamin, Kirsten R.; Silva, Iris R.; Cherubim, João P.; McPhee, Derek J.; Paddon, Christopher J.

doi:10.5935/0103-5053.20160119

Cited by 36 publications

(51 citation statements)

References 27 publications

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“…The most well-studied genetic switches rely on the addition of galactose to the fermentation medium, which is expensive and would add to the production cost. There has been considerable progress in the industrial production of terpenes by yeast in the decade following the amorphadiene work described above, though directed primarily at production of another sesquiterpene, β-farnesene ( Benjamin et al, 2016 ; Leavell et al, 2016 ).…”

Section: Developing Terpenoid Production Directly From Sugarmentioning

confidence: 99%

Approaches and Recent Developments for the Commercial Production of Semi-synthetic Artemisinin

et al. 2018

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The antimalarial drug artemisinin is a natural product produced by the plant Artemisia annua. Extracts of A. annua have been used in Chinese herbal medicine for over two millennia. Following the re-discovery of A. annua extract as an effective antimalarial, and the isolation and structural elucidation of artemisinin as the active agent, it was recommended as the first-line treatment for uncomplicated malaria in combination with another effective antimalarial drug (Artemisinin Combination Therapy) by the World Health Organization (WHO) in 2002. Following the WHO recommendation, the availability and price of artemisinin fluctuated greatly, ranging from supply shortfalls in some years to oversupply in others. To alleviate these supply and price issues, a second source of artemisinin was sought, resulting in an effort to produce artemisinic acid, a late-stage chemical precursor of artemisinin, by yeast fermentation, followed by chemical conversion to artemisinin (i.e., semi-synthesis). Engineering to enable production of artemisinic acid in yeast relied on the discovery of A. annua genes encoding artemisinic acid biosynthetic enzymes, and synthetic biology to engineer yeast metabolism. The progress of this effort, which resulted in semi-synthetic artemisinin entering commercial production in 2013, is reviewed with an emphasis on recent publications and opportunities for further development. Aspects of both the biology of artemisinin production in A. annua, and yeast strain engineering are discussed, as are recent developments in the chemical conversion of artemisinic acid to artemisinin.

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Section: Developing Terpenoid Production Directly From Sugarmentioning

confidence: 99%

Approaches and Recent Developments for the Commercial Production of Semi-synthetic Artemisinin

et al. 2018

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“…Isoprenoids are a promising class of molecules with over 40,000 known structures and potential uses as pharmaceuticals, flavors, fragrances, pesticides, disinfectants, and chemical feedstocks (Bohlmann and Keeling, 2008;George et al, 2015;Jongedijk et al, 2016). While there have been demonstrations of cellular isoprenoid production at commercial scales (Benjamin et al, 2016;Paddon et al, 2013), efforts to engineer strains for new products have proved challenging.…”

Section: Introductionmentioning

confidence: 99%

Cell-free prototyping of limonene biosynthesis using cell-free protein synthesis

Dudley¹,

Karim²,

Nash³

et al. 2020

Preprint

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Metabolic engineering of microorganisms to produce sustainable chemicals has emerged as an important part of the global bioeconomy. Unfortunately, efforts to design and engineer microbial cell factories are challenging because design-built-test cycles, iterations of re-engineering organisms to test and optimize new sets of enzymes, are slow. To alleviate this challenge, we demonstrate a cell-free approach termed in vitro Prototyping and Rapid Optimization of Biosynthetic Enzymes (or iPROBE). In iPROBE, a large number of pathway combinations can be rapidly built and optimized. The key idea is to use cell-free protein synthesis (CFPS) to manufacture pathway enzymes in separate reactions that are then mixed to modularly assemble multiple, distinct biosynthetic pathways. As a model, we apply our approach to the 9-step heterologous enzyme pathway to limonene in extracts from Escherichia coli. In iterative cycles of design, we studied the impact of 54 enzyme homologs, multiple enzyme levels, and cofactor concentrations on pathway performance. In total, we screened over 150 unique sets of enzymes in 580 unique pathway conditions to increase limonene production in 24 hours from 0.2 to 4.5 mM (23 to 610 mg/L). Finally, to demonstrate the modularity of this pathway, we also synthesized the biofuel precursors pinene and bisabolene. We anticipate that iPROBE will accelerate design-build-test cycles for metabolic engineering, enabling data-driven multiplexed cell-free methods for testing large combinations of biosynthetic enzymes to inform cellular design. Dudley, et al. -Cell-free prototyping of limonene biosynthesis 3 TOC Figure Highlights • Applied the iPROBE framework to build the nine-enzyme pathway to produce limonene • Assessed the impact of cofactors and 54 enzyme homologs on cell-free enzyme performance • Iteratively optimized the cell-free production of limonene by exploring more than 580 unique reactions • Extended pathway to biofuel precursors pinene and bisabolene Dudley, et al. -Cell-free prototyping of limonene biosynthesis 4

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“…Other efforts to enhance artemisinin production have been attempted through genetic engineering of the artemisinin biosynthetic pathway genes in microbial heterologous hosts. Extensive work on the development of microbial production of artemisinin precursors led to semi-synthesis of artemisinin, but this is only partly commercially successful ( Benjamin et al, 2016 ; Peplow, 2016 ; Singh et al, 2017 ). In this review, the progress and recent bioengineering advances in artemisinin production in stable heterologous in planta systems including genetic modifications of A. annua is summarized.…”

Section: Introductionmentioning

confidence: 99%

A Review of Biotechnological Artemisinin Production in Plants

Ikram

Simonsen

2017

Front. Plant Sci.

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Malaria is still an eminent threat to major parts of the world population mainly in sub-Saharan Africa. Researchers around the world continuously seek novel solutions to either eliminate or treat the disease. Artemisinin, isolated from the Chinese medicinal herb Artemisia annua, is the active ingredient in artemisinin-based combination therapies used to treat the disease. However, naturally artemisinin is produced in small quantities, which leads to a shortage of global supply. Due to its complex structure, it is difficult chemically synthesize. Thus to date, A. annua remains as the main commercial source of artemisinin. Current advances in genetic and metabolic engineering drives to more diverse approaches and developments on improving in planta production of artemisinin, both in A. annua and in other plants. In this review, we describe efforts in bioengineering to obtain a higher production of artemisinin in A. annua and stable heterologous in planta systems. The current progress and advancements provides hope for significantly improved production in plants.

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Developing Commercial Production of Semi-Synthetic Artemisinin, and of β-Farnesene, an Isoprenoid Produced by Fermentation of Brazilian Sugar

Cited by 36 publications

References 27 publications

Approaches and Recent Developments for the Commercial Production of Semi-synthetic Artemisinin

Approaches and Recent Developments for the Commercial Production of Semi-synthetic Artemisinin

Cell-free prototyping of limonene biosynthesis using cell-free protein synthesis

A Review of Biotechnological Artemisinin Production in Plants

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