The Experts below are selected from a list of 1221 Experts worldwide ranked by ideXlab platform
Christopher J. Paddon - One of the best experts on this subject based on the ideXlab platform.
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amalgamation of synthetic biology and chemistry for high throughput nonconventional synthesis of the antimalarial drug artemisinin
Organic Process Research & Development, 2017Co-Authors: Dharmendra Singh, Christopher J. Paddon, Derek Mcphee, Joel Cherry, Ghanshyam Maurya, Ganesh Mahale, Yogesh Patel, Neeraj Kumar, Subhash C Singh, Brajesh SharmaAbstract:The development of a cost-effective process for the production of artemisinin, the precursor of all artemisinin-derived drugs, the first-line treatment for malaria, has been a long-pursued endeavor. The breakthrough achievement of coaxing genetically engineered yeast to express Artemisia annua genes for the commercial production of Artemisinic Acid, an advanced intermediate in the synthesis of artemisinin, has yet to fully realize an affordable malaria treatment for the poor because of the lack of a cost-effective chemical conversion into artemisinin. We describe herein a commercially feasible and pragmatic synthesis of artemisinin from amorpha-4,11-diene, an early-stage intermediate produced in 2-fold higher molar yield than engineered yeast cells can process into Artemisinic Acid. The key to this novel approach is an exceedingly effective functionalization of the isopropenyl group of amorphadiene via endo-epoxyamorphadiene to give dihydroArtemisinic Acid, which upon esterification followed by oxidation ...
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Amalgamation of Synthetic Biology and Chemistry for High-Throughput Nonconventional Synthesis of the Antimalarial Drug Artemisinin
2017Co-Authors: Dharmendra Singh, Christopher J. Paddon, Derek Mcphee, Joel Cherry, Ghanshyam Maurya, Ganesh Mahale, Yogesh Patel, Neeraj Kumar, Subhash Singh, Brajesh SharmaAbstract:The development of a cost-effective process for the production of artemisinin, the precursor of all artemisinin-derived drugs, the first-line treatment for malaria, has been a long-pursued endeavor. The breakthrough achievement of coaxing genetically engineered yeast to express Artemisia annua genes for the commercial production of Artemisinic Acid, an advanced intermediate in the synthesis of artemisinin, has yet to fully realize an affordable malaria treatment for the poor because of the lack of a cost-effective chemical conversion into artemisinin. We describe herein a commercially feasible and pragmatic synthesis of artemisinin from amorpha-4,11-diene, an early-stage intermediate produced in 2-fold higher molar yield than engineered yeast cells can process into Artemisinic Acid. The key to this novel approach is an exceedingly effective functionalization of the isopropenyl group of amorphadiene via endo-epoxyamorphadiene to give dihydroArtemisinic Acid, which upon esterification followed by oxidation and cyclicization furnishes pure artemisinin in approximately 60% yield
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high level semi synthetic production of the potent antimalarial artemisinin
Nature, 2013Co-Authors: Christopher J. Paddon, Douglas J. Pitera, Kirsten R Benjamin, Karl Fisher, Patrick J Westfall, Michael D. Leavell, Derek James Mcphee, A Main, Devin R Polichuk, Keat Thomas H TeohAbstract:Saccharomyces cerevisiae is engineered to produce high concentrations of Artemisinic Acid, a precursor of the artemisinin used in combination therapies for malaria treatment; an efficient and practical chemical process to convert Artemisinic Acid to artemisinin is also developed. Artemisinin-based combination therapies are the treatment of choice for uncomplicated Plasmodium falciparum malaria, but the supply of plant-derived artemisinin can sometimes be unreliable, causing shortages and high prices. This manuscript describes a viable industrial process for the production of semisynthetic artemisinin, with the potential to help stabilize artemisinin supply. The process uses Saccharomyces cerevisiae yeast engineered to produce high yields of Artemisinic Acid, a precursor of artemisinin. The authors have also developed an efficient and scalable chemical process to convert Artemisinic Acid to artemisinin. In 2010 there were more than 200 million cases of malaria, and at least 655,000 deaths1. The World Health Organization has recommended artemisinin-based combination therapies (ACTs) for the treatment of uncomplicated malaria caused by the parasite Plasmodium falciparum. Artemisinin is a sesquiterpene endoperoxide with potent antimalarial properties, produced by the plant Artemisia annua. However, the supply of plant-derived artemisinin is unstable, resulting in shortages and price fluctuations, complicating production planning by ACT manufacturers2. A stable source of affordable artemisinin is required. Here we use synthetic biology to develop strains of Saccharomyces cerevisiae (baker’s yeast) for high-yielding biological production of Artemisinic Acid, a precursor of artemisinin. Previous attempts to produce commercially relevant concentrations of Artemisinic Acid were unsuccessful, allowing production of only 1.6 grams per litre of Artemisinic Acid3. Here we demonstrate the complete biosynthetic pathway, including the discovery of a plant dehydrogenase and a second cytochrome that provide an efficient biosynthetic route to Artemisinic Acid, with fermentation titres of 25 grams per litre of Artemisinic Acid. Furthermore, we have developed a practical, efficient and scalable chemical process for the conversion of Artemisinic Acid to artemisinin using a chemical source of singlet oxygen, thus avoiding the need for specialized photochemical equipment. The strains and processes described here form the basis of a viable industrial process for the production of semi-synthetic artemisinin to stabilize the supply of artemisinin for derivatization into active pharmaceutical ingredients (for example, artesunate) for incorporation into ACTs. Because all intellectual property rights have been provided free of charge, this technology has the potential to increase provision of first-line antimalarial treatments to the developing world at a reduced average annual price.
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high level semi synthetic production of the potent antimalarial artemisinin
Nature, 2013Co-Authors: Christopher J. Paddon, Douglas J. Pitera, Kirsten R Benjamin, Karl Fisher, Patrick J Westfall, Michael D. Leavell, Derek James Mcphee, A Main, Anna Tai, Diana EngAbstract:Saccharomyces cerevisiae is engineered to produce high concentrations of Artemisinic Acid, a precursor of the artemisinin used in combination therapies for malaria treatment; an efficient and practical chemical process to convert Artemisinic Acid to artemisinin is also developed.
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Microbially Derived Semisynthetic Artemisinin
Isoprenoid Synthesis in Plants and Microorganisms, 2012Co-Authors: Christopher J. Paddon, Scott Fickes, Douglas J. Pitera, Rika Regentin, Kirsten R Benjamin, Karl Fisher, Patrick J Westfall, Michael D. Leavell, Derek James Mcphee, Jack D NewmanAbstract:Artemisinin is a sesquiterpene lactone endoperoxide with potent antimalarial properties, recommended by the World Health Organization for the treatment of malaria in artemisinin combination therapies (ACTs). It is extracted from the plant Artemisia annua, but its supplies are limited and its price is volatile. In order to increase supply and stabilize the price of artemisinin, a semisynthesis has been developed, whereby an artemisinin precursor (amorpha-4,11-diene) is produced in microbes and the isolated precursor converted chemically to artemisinin. Escherichia coli has been engineered to produce amorpha-4,11-diene by the expression of a heterologous mevalonate pathway along with amorpha-4,11-diene synthase (ADS) from A. annua. Development of the E. coli platform to increase production of amorpha-4,11-diene from 24 mg/L to >25 g/L is described. ADS has also been expressed in the yeast model system Saccharomyces cerevisiae which, following manipulation of the mevalonate pathway, produced 150 mg/L of amorpha-4,11-diene. The cDNAs encoding the cytochrome P450 that oxidizes amorpha-4,11-diene to Artemisinic Acid, CYP71AV1, and its cognate reductase were isolated from A. annua and expressed in amorpha-4,11-diene-producing E. coli and yeast, leading to the production of >1 g/L Artemisinic Acid from both organisms. A route for the chemical conversion of Artemisinic Acid to artemisinin is described. Production of semisynthetic artemisinin may lead to the development of a second source of the drug for incorporation into ACTs.
Jean-luc Veuthey - One of the best experts on this subject based on the ideXlab platform.
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Extraction of artemisinin and Artemisinic Acid from Artemisia annua L. using supercritical carbon dioxide
Journal of chromatography. A, 1997Co-Authors: Marcel Kohler, Werner Haerdi, Philippe Christen, Jean-luc VeutheyAbstract:Artemisinin (an antimalaric compound) and its major precursor Artemisinic Acid, isolated as the active principles of the medicinal plant Artemisia annua L., were extracted by supercritical fluid extraction (SFE) and analyzed by supercritical fluid chromatography (SFC) using a capillary column, coupled with a flame ionization detector (FID). With optimized operating conditions, artemisinin and Artemisinic Acid were quantitatively extracted at a flow-rate of 2 ml min-1 in less than 20 min. The supercritical fluid was composed of carbon dioxide and 3% methanol with temperature and pressure fixed at 50 degrees C and 15 MPa, respectively. From the kinetic curves, it appears that the extraction of artemisinin is not limited by the diffusion of the analyte from the plant into the extraction fluid but rather by the elution process. These conditions avoided degradation of the analyte and gave clean extracts ready to be analyzed by SFC. The SFE-SFC-FID method was successfully applied to six samples of A. annua containing various concentrations of artemisinin and Artemisinic Acid. Results were compared with two conventional liquid solvent extraction processes.
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Determination of artemisinin and Artemisinic Acid by capillary and packed supercritical fluid chromatography
Journal of High Resolution Chromatography, 1997Co-Authors: Marcel Kohler, Werner Haerdi, Philippe Christen, Jean-luc VeutheyAbstract:Artemisinin (an antimalarial compound) and its bioprecursor Artemisinic Acid, present in the plant Atemisia annua L., were analyzed by supercritical fluid chromatography (SFS) using capillary and packed columns, coupled respectively with a flame ionization detector (FID) and an evaporative light scattering detector (ELSD). Both methods were optimized and validated with columns of different polarity in order to separate artemisinin and Artemisinic Acid. Analytical results were comparable, but the paced SFC-ELSD method was faster. Indeed, artemisinin and Artemisinic Acid were separated with an aminopropyl silica column in less than 8 minutes instead of about 25 minutes by capillary SFS. Contrary to conventional gas and liquid chromatography coupled to an UV-visible detector, SFS methods determined both compounds directly, without degradation and/or derivatization in the concentration range expected in the plant material. Results obtained on plant extracts by capillary SFS-FID and packed SFS-ELSD were confirmed by GC-MS.
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Supercritical fluid extraction and chromatography of artemisinin and Artemisinic Acid. An improved method for the analysis of Artemisia annua samples
Phytochemical Analysis, 1997Co-Authors: Marcel Kohler, Werner Haerdi, Philippe Christen, Jean-luc VeutheyAbstract:Artemisinin and Artemisinic Acid were extracted from aerial parts of Artemisia annua by supercritical fluid extraction (SFE) with carbon dioxide modified with 3% (v/v) methanol. The quantitative determination of both compounds was carried out by supercritical fluid chromatography (SFC) coupled with an evaporative light scattering detector (SFC-ELSD). Results obtained by SFE-SFC-ELSD show that this method allows mild extraction conditions and quantitative determination without further purification of the plant extract. In all cases, artemisinin and Artemisinic Acid were extracted in a higher yield with supercritical carbon dioxide than with liquid-solid extraction. © 1997 John Wiley & Sons, Ltd.
Patrick J Westfall - One of the best experts on this subject based on the ideXlab platform.
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high level semi synthetic production of the potent antimalarial artemisinin
Nature, 2013Co-Authors: Christopher J. Paddon, Douglas J. Pitera, Kirsten R Benjamin, Karl Fisher, Patrick J Westfall, Michael D. Leavell, Derek James Mcphee, A Main, Devin R Polichuk, Keat Thomas H TeohAbstract:Saccharomyces cerevisiae is engineered to produce high concentrations of Artemisinic Acid, a precursor of the artemisinin used in combination therapies for malaria treatment; an efficient and practical chemical process to convert Artemisinic Acid to artemisinin is also developed. Artemisinin-based combination therapies are the treatment of choice for uncomplicated Plasmodium falciparum malaria, but the supply of plant-derived artemisinin can sometimes be unreliable, causing shortages and high prices. This manuscript describes a viable industrial process for the production of semisynthetic artemisinin, with the potential to help stabilize artemisinin supply. The process uses Saccharomyces cerevisiae yeast engineered to produce high yields of Artemisinic Acid, a precursor of artemisinin. The authors have also developed an efficient and scalable chemical process to convert Artemisinic Acid to artemisinin. In 2010 there were more than 200 million cases of malaria, and at least 655,000 deaths1. The World Health Organization has recommended artemisinin-based combination therapies (ACTs) for the treatment of uncomplicated malaria caused by the parasite Plasmodium falciparum. Artemisinin is a sesquiterpene endoperoxide with potent antimalarial properties, produced by the plant Artemisia annua. However, the supply of plant-derived artemisinin is unstable, resulting in shortages and price fluctuations, complicating production planning by ACT manufacturers2. A stable source of affordable artemisinin is required. Here we use synthetic biology to develop strains of Saccharomyces cerevisiae (baker’s yeast) for high-yielding biological production of Artemisinic Acid, a precursor of artemisinin. Previous attempts to produce commercially relevant concentrations of Artemisinic Acid were unsuccessful, allowing production of only 1.6 grams per litre of Artemisinic Acid3. Here we demonstrate the complete biosynthetic pathway, including the discovery of a plant dehydrogenase and a second cytochrome that provide an efficient biosynthetic route to Artemisinic Acid, with fermentation titres of 25 grams per litre of Artemisinic Acid. Furthermore, we have developed a practical, efficient and scalable chemical process for the conversion of Artemisinic Acid to artemisinin using a chemical source of singlet oxygen, thus avoiding the need for specialized photochemical equipment. The strains and processes described here form the basis of a viable industrial process for the production of semi-synthetic artemisinin to stabilize the supply of artemisinin for derivatization into active pharmaceutical ingredients (for example, artesunate) for incorporation into ACTs. Because all intellectual property rights have been provided free of charge, this technology has the potential to increase provision of first-line antimalarial treatments to the developing world at a reduced average annual price.
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high level semi synthetic production of the potent antimalarial artemisinin
Nature, 2013Co-Authors: Christopher J. Paddon, Douglas J. Pitera, Kirsten R Benjamin, Karl Fisher, Patrick J Westfall, Michael D. Leavell, Derek James Mcphee, A Main, Anna Tai, Diana EngAbstract:Saccharomyces cerevisiae is engineered to produce high concentrations of Artemisinic Acid, a precursor of the artemisinin used in combination therapies for malaria treatment; an efficient and practical chemical process to convert Artemisinic Acid to artemisinin is also developed.
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Microbially Derived Semisynthetic Artemisinin
Isoprenoid Synthesis in Plants and Microorganisms, 2012Co-Authors: Christopher J. Paddon, Scott Fickes, Douglas J. Pitera, Rika Regentin, Kirsten R Benjamin, Karl Fisher, Patrick J Westfall, Michael D. Leavell, Derek James Mcphee, Jack D NewmanAbstract:Artemisinin is a sesquiterpene lactone endoperoxide with potent antimalarial properties, recommended by the World Health Organization for the treatment of malaria in artemisinin combination therapies (ACTs). It is extracted from the plant Artemisia annua, but its supplies are limited and its price is volatile. In order to increase supply and stabilize the price of artemisinin, a semisynthesis has been developed, whereby an artemisinin precursor (amorpha-4,11-diene) is produced in microbes and the isolated precursor converted chemically to artemisinin. Escherichia coli has been engineered to produce amorpha-4,11-diene by the expression of a heterologous mevalonate pathway along with amorpha-4,11-diene synthase (ADS) from A. annua. Development of the E. coli platform to increase production of amorpha-4,11-diene from 24 mg/L to >25 g/L is described. ADS has also been expressed in the yeast model system Saccharomyces cerevisiae which, following manipulation of the mevalonate pathway, produced 150 mg/L of amorpha-4,11-diene. The cDNAs encoding the cytochrome P450 that oxidizes amorpha-4,11-diene to Artemisinic Acid, CYP71AV1, and its cognate reductase were isolated from A. annua and expressed in amorpha-4,11-diene-producing E. coli and yeast, leading to the production of >1 g/L Artemisinic Acid from both organisms. A route for the chemical conversion of Artemisinic Acid to artemisinin is described. Production of semisynthetic artemisinin may lead to the development of a second source of the drug for incorporation into ACTs.
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production of amorphadiene in yeast and its conversion to dihydroArtemisinic Acid precursor to the antimalarial agent artemisinin
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Patrick J Westfall, Douglas J. Pitera, Rika Regentin, Jacob R Lenihan, Hiroko Tsuruta, Frank X Woolard, Tizita Horning, David J Melis, Andrew Owens, Scott FickesAbstract:Malaria, caused by Plasmodium sp, results in almost one million deaths and over 200 million new infections annually. The World Health Organization has recommended that artemisinin-based combination therapies be used for treatment of malaria. Artemisinin is a sesquiterpene lactone isolated from the plant Artemisia annua. However, the supply and price of artemisinin fluctuate greatly, and an alternative production method would be valuable to increase availability. We describe progress toward the goal of developing a supply of semisynthetic artemisinin based on production of the artemisinin precursor amorpha-4,11-diene by fermentation from engineered Saccharomyces cerevisiae, and its chemical conversion to dihydroArtemisinic Acid, which can be subsequently converted to artemisinin. Previous efforts to produce artemisinin precursors used S. cerevisiae S288C overexpressing selected genes of the mevalonate pathway [Ro et al. (2006) Nature 440:940–943]. We have now overexpressed every enzyme of the mevalonate pathway to ERG20 in S. cerevisiae CEN.PK2, and compared production to CEN.PK2 engineered identically to the previously engineered S288C strain. Overexpressing every enzyme of the mevalonate pathway doubled Artemisinic Acid production, however, amorpha-4,11-diene production was 10-fold higher than Artemisinic Acid. We therefore focused on amorpha-4,11-diene production. Development of fermentation processes for the reengineered CEN.PK2 amorpha-4,11-diene strain led to production of > 40 g/L product. A chemical process was developed to convert amorpha-4,11-diene to dihydroArtemisinic Acid, which could subsequently be converted to artemisinin. The strains and procedures described represent a complete process for production of semisynthetic artemisinin.
Douglas J. Pitera - One of the best experts on this subject based on the ideXlab platform.
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high level semi synthetic production of the potent antimalarial artemisinin
Nature, 2013Co-Authors: Christopher J. Paddon, Douglas J. Pitera, Kirsten R Benjamin, Karl Fisher, Patrick J Westfall, Michael D. Leavell, Derek James Mcphee, A Main, Devin R Polichuk, Keat Thomas H TeohAbstract:Saccharomyces cerevisiae is engineered to produce high concentrations of Artemisinic Acid, a precursor of the artemisinin used in combination therapies for malaria treatment; an efficient and practical chemical process to convert Artemisinic Acid to artemisinin is also developed. Artemisinin-based combination therapies are the treatment of choice for uncomplicated Plasmodium falciparum malaria, but the supply of plant-derived artemisinin can sometimes be unreliable, causing shortages and high prices. This manuscript describes a viable industrial process for the production of semisynthetic artemisinin, with the potential to help stabilize artemisinin supply. The process uses Saccharomyces cerevisiae yeast engineered to produce high yields of Artemisinic Acid, a precursor of artemisinin. The authors have also developed an efficient and scalable chemical process to convert Artemisinic Acid to artemisinin. In 2010 there were more than 200 million cases of malaria, and at least 655,000 deaths1. The World Health Organization has recommended artemisinin-based combination therapies (ACTs) for the treatment of uncomplicated malaria caused by the parasite Plasmodium falciparum. Artemisinin is a sesquiterpene endoperoxide with potent antimalarial properties, produced by the plant Artemisia annua. However, the supply of plant-derived artemisinin is unstable, resulting in shortages and price fluctuations, complicating production planning by ACT manufacturers2. A stable source of affordable artemisinin is required. Here we use synthetic biology to develop strains of Saccharomyces cerevisiae (baker’s yeast) for high-yielding biological production of Artemisinic Acid, a precursor of artemisinin. Previous attempts to produce commercially relevant concentrations of Artemisinic Acid were unsuccessful, allowing production of only 1.6 grams per litre of Artemisinic Acid3. Here we demonstrate the complete biosynthetic pathway, including the discovery of a plant dehydrogenase and a second cytochrome that provide an efficient biosynthetic route to Artemisinic Acid, with fermentation titres of 25 grams per litre of Artemisinic Acid. Furthermore, we have developed a practical, efficient and scalable chemical process for the conversion of Artemisinic Acid to artemisinin using a chemical source of singlet oxygen, thus avoiding the need for specialized photochemical equipment. The strains and processes described here form the basis of a viable industrial process for the production of semi-synthetic artemisinin to stabilize the supply of artemisinin for derivatization into active pharmaceutical ingredients (for example, artesunate) for incorporation into ACTs. Because all intellectual property rights have been provided free of charge, this technology has the potential to increase provision of first-line antimalarial treatments to the developing world at a reduced average annual price.
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high level semi synthetic production of the potent antimalarial artemisinin
Nature, 2013Co-Authors: Christopher J. Paddon, Douglas J. Pitera, Kirsten R Benjamin, Karl Fisher, Patrick J Westfall, Michael D. Leavell, Derek James Mcphee, A Main, Anna Tai, Diana EngAbstract:Saccharomyces cerevisiae is engineered to produce high concentrations of Artemisinic Acid, a precursor of the artemisinin used in combination therapies for malaria treatment; an efficient and practical chemical process to convert Artemisinic Acid to artemisinin is also developed.
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Microbially Derived Semisynthetic Artemisinin
Isoprenoid Synthesis in Plants and Microorganisms, 2012Co-Authors: Christopher J. Paddon, Scott Fickes, Douglas J. Pitera, Rika Regentin, Kirsten R Benjamin, Karl Fisher, Patrick J Westfall, Michael D. Leavell, Derek James Mcphee, Jack D NewmanAbstract:Artemisinin is a sesquiterpene lactone endoperoxide with potent antimalarial properties, recommended by the World Health Organization for the treatment of malaria in artemisinin combination therapies (ACTs). It is extracted from the plant Artemisia annua, but its supplies are limited and its price is volatile. In order to increase supply and stabilize the price of artemisinin, a semisynthesis has been developed, whereby an artemisinin precursor (amorpha-4,11-diene) is produced in microbes and the isolated precursor converted chemically to artemisinin. Escherichia coli has been engineered to produce amorpha-4,11-diene by the expression of a heterologous mevalonate pathway along with amorpha-4,11-diene synthase (ADS) from A. annua. Development of the E. coli platform to increase production of amorpha-4,11-diene from 24 mg/L to >25 g/L is described. ADS has also been expressed in the yeast model system Saccharomyces cerevisiae which, following manipulation of the mevalonate pathway, produced 150 mg/L of amorpha-4,11-diene. The cDNAs encoding the cytochrome P450 that oxidizes amorpha-4,11-diene to Artemisinic Acid, CYP71AV1, and its cognate reductase were isolated from A. annua and expressed in amorpha-4,11-diene-producing E. coli and yeast, leading to the production of >1 g/L Artemisinic Acid from both organisms. A route for the chemical conversion of Artemisinic Acid to artemisinin is described. Production of semisynthetic artemisinin may lead to the development of a second source of the drug for incorporation into ACTs.
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production of amorphadiene in yeast and its conversion to dihydroArtemisinic Acid precursor to the antimalarial agent artemisinin
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Patrick J Westfall, Douglas J. Pitera, Rika Regentin, Jacob R Lenihan, Hiroko Tsuruta, Frank X Woolard, Tizita Horning, David J Melis, Andrew Owens, Scott FickesAbstract:Malaria, caused by Plasmodium sp, results in almost one million deaths and over 200 million new infections annually. The World Health Organization has recommended that artemisinin-based combination therapies be used for treatment of malaria. Artemisinin is a sesquiterpene lactone isolated from the plant Artemisia annua. However, the supply and price of artemisinin fluctuate greatly, and an alternative production method would be valuable to increase availability. We describe progress toward the goal of developing a supply of semisynthetic artemisinin based on production of the artemisinin precursor amorpha-4,11-diene by fermentation from engineered Saccharomyces cerevisiae, and its chemical conversion to dihydroArtemisinic Acid, which can be subsequently converted to artemisinin. Previous efforts to produce artemisinin precursors used S. cerevisiae S288C overexpressing selected genes of the mevalonate pathway [Ro et al. (2006) Nature 440:940–943]. We have now overexpressed every enzyme of the mevalonate pathway to ERG20 in S. cerevisiae CEN.PK2, and compared production to CEN.PK2 engineered identically to the previously engineered S288C strain. Overexpressing every enzyme of the mevalonate pathway doubled Artemisinic Acid production, however, amorpha-4,11-diene production was 10-fold higher than Artemisinic Acid. We therefore focused on amorpha-4,11-diene production. Development of fermentation processes for the reengineered CEN.PK2 amorpha-4,11-diene strain led to production of > 40 g/L product. A chemical process was developed to convert amorpha-4,11-diene to dihydroArtemisinic Acid, which could subsequently be converted to artemisinin. The strains and procedures described represent a complete process for production of semisynthetic artemisinin.
Rajendra Singh Bhakuni - One of the best experts on this subject based on the ideXlab platform.
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biotransformation of Artemisinic Acid by the fungus trichothecium roseum and anti candidal activity of its metabolites
Biocatalysis and Biotransformation, 2019Co-Authors: Pooja Singh, Deepak Singh Kapkoti, Rashi Tewari, Prakash Kumar Rout, N P Singh, Dharmendra Saikia, Rakesh Pandey, Rajendra Singh BhakuniAbstract:AbstractThe microbial transformation of Artemisinic Acid (1) using cell culture of endophytic fungus Trichothecium roseum was investigated. Previously, we have reported two major metabolites, 3β-hydroxyArtemisinic Acid (2) and 3β,15-dihydroxyArtemisinic Acid (3) from the biotransformation of Artemisinic Acid by the fungus T. roseum CIMAPN1. Here in the present paper, we obtained a new minor compound 4 (5.2% in yield) along with compounds 2 and 3 through scale-up of biotransformation process of Artemisinic Acid using the same fungus. The structure of compound 4 was established as 3-oxoArtemisinic Acid on the basis of its IR, ESI-MS, HRMS, 1 D (1H and 13C, DEPT), and 2 D (COSY, HSQC, HMBC) NMR spectral data analysis. The possible reaction mechanism of the formation of 3-oxoArtemisinic Acid from Artemisinic Acid was proposed. Furthermore, all the three metabolites along with the Artemisinic Acid were evaluated for their antifungal activity against the three fungal strains Candida albicans (ATCC 14053), Candi...
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Biotransformation of Artemisinic Acid by the fungus Trichothecium roseum and anti-candidal activity of its metabolites
Biocatalysis and Biotransformation, 2019Co-Authors: Pooja Singh, Deepak Singh Kapkoti, Rashi Tewari, Dharmendra Saikia, Rakesh Pandey, Nandan Singh, Prasant Kumar Rout, Rajendra Singh BhakuniAbstract:The microbial transformation of Artemisinic Acid (1) using cell culture of endophytic fungus Trichothecium roseum was investigated. Previously, we have reported two major metabolites, 3β-hydroxyart...
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Biotransformation of Artemisinic Acid by the fungus Trichothecium roseum and anti-candidal activity of its metabolites
2019Co-Authors: Pooja Singh, Deepak Singh Kapkoti, Rashi Tewari, Dharmendra Saikia, Rakesh Pandey, Nandan Singh, Prasant Kumar Rout, Rajendra Singh BhakuniAbstract:The microbial transformation of Artemisinic Acid (1) using cell culture of endophytic fungus Trichothecium roseum was investigated. Previously, we have reported two major metabolites, 3β-hydroxyArtemisinic Acid (2) and 3β,15-dihydroxyArtemisinic Acid (3) from the biotransformation of Artemisinic Acid by the fungus T. roseum CIMAPN1. Here in the present paper, we obtained a new minor compound 4 (5.2% in yield) along with compounds 2 and 3 through scale-up of biotransformation process of Artemisinic Acid using the same fungus. The structure of compound 4 was established as 3-oxoArtemisinic Acid on the basis of its IR, ESI-MS, HRMS, 1 D (1H and 13C, DEPT), and 2 D (COSY, HSQC, HMBC) NMR spectral data analysis. The possible reaction mechanism of the formation of 3-oxoArtemisinic Acid from Artemisinic Acid was proposed. Furthermore, all the three metabolites along with the Artemisinic Acid were evaluated for their antifungal activity against the three fungal strains Candida albicans (ATCC 14053), Candida albicans clinical isolates and Candida kefyr (ATCC 204093). 3-OxoArtemisinic Acid was the most active (4 to 16 times more potent than Artemisinic Acid) with MIC ranges from 125 to 500 µg/mL among all tested compounds. This study suggested that the Artemisinic Acid molecule has a great potential to be exploited for further biotransformation by the different fungi and can produce chemically diverse molecules with better biological activity.
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novel biotransformation processes of Artemisinic Acid to their hydroxylated derivatives 3β hydroxyArtemisinic Acid and 3β 15 dihydroxyArtemisinic by fungus trichothecium roseum cimapn1and their biological evaluation
Journal of Molecular Catalysis B-enzymatic, 2014Co-Authors: Rashmi Gaur, Rakesh Pandey, Sudeep Tiwari, Apurva Jakhmola, Jay Prakash Thakur, Ram Kishor Verma, Rajendra Singh BhakuniAbstract:Abstract The biotransformation of Artemisinic Acid ( 1 ) by endophytic fungus Trichothecium roseum CIMAPN1 is reported here for the first time. The major biotransformed products appeared as a grayish color spot on thin-layer chromatography (TLC) with transparent crystal-like texture. Based on their infrared (IR) and 1 H nuclear magnetic resonance (NMR) spectra, the products were characterized as a 3β-hydroxyArtemisinic Acid ( 2) and 3β, 15-dihydroxyArtemisinic Acid ( 3) and were obtained in 51.1% and 37.3% yields, respectively. The highest conversion efficiencies were obtained respectively when 2-day-old cultures of T. roseum were fed with 20 mg of compound 1 in 50 ml of medium per culture and the mycelia were harvested after 14 days of incubation. Metabolite 3 is a new compound and metabolite 2 is reported here for the first time from Artemisinic Acid. Compound 3 was acetylated to the product diacetate derivative 4 . All these compounds were evaluated for their antimicrobial and in vitro antioxidant activities. Further, for pharmaceutical utility these compounds were analyzed in vivo using well established animal model Caenorhabditis elegans for antioxidant/reactive oxygen species scavenging activity. The results clearly showed that the novel derivatives performed well in comparison to the parent compound both in-vitro and in-vivo studies. The presence of hydroxyl groups in metabolites 2 and 3 could make them interesting synthones for further modification into new clinically potent molecules. Thus, the study suggested a new aisle towards better drug development through the utilization of micro engineers.
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Isomeric Flavonoids of Artemisia annua (Asterales: Asteraceae) as Insect Growth Inhibitors Against Helicoverpa armigera (Lepidoptera: Noctuidae)
Florida Entomologist, 2013Co-Authors: Neelima Anshul, Rajendra Singh Bhakuni, Rashmi Gaur, Dwijendra SinghAbstract:ABSTRACT Artemisia annua (Asterales: Asteraceae) is one of the important natural sources of antimalarial compounds i.e., artemisinin and Artemisinic Acid. Also this plant is cultivated on a large area in India under industry-farmer partnerships. With a view to enhance the added value of the raw material of A. annua and its chemical constituents, we evaluated methanolic extract of powdered A. annua leaves and different compounds isolated from the extract for toxicity and inhibition and disruption of growth and development of the African pod borer, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae). Methanol extract of A. annua and eight known constituent compounds [Artemisinic Acid, artemisinin, scopoletin, arteannuin-B, deoxy-artemisinin, artemetin and isomeric flavonoids (casticin and chrysosplenetin)] were bio-assayed for larval mortality, abnormal development, and growth inhibition. The methanol extract severely affected 100% of the larva treated, i.e., larvae gained very little weight, some larvae...