The Experts below are selected from a list of 56253 Experts worldwide ranked by ideXlab platform
Meng Wang - One of the best experts on this subject based on the ideXlab platform.
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Discovery and Engineering of Cytochrome P450s for Terpenoid Biosynthesis.
Trends in biotechnology, 2018Co-Authors: Han Xiao, Yue Zhang, Meng WangAbstract:Terpenoids represent 60% of known natural products, including many drugs and drug candidates, and their biosynthesis is attracting great interest. However, the unknown cytochrome P450s (CYPs) in Terpenoid biosynthetic pathways make the heterologous production of related Terpenoids impossible, while the slow kinetics of some known CYPs greatly limit the efficiency of Terpenoid biosynthesis. Thus, there is a compelling need to discover and engineer CYPs for Terpenoid biosynthesis to fully realize their great potential for industrial application. This review article summarizes the current state of CYP discovery and engineering in Terpenoid biosynthesis, focusing on recent synthetic biology approaches toward prototyping CYPs in heterologous hosts. We also propose several strategies for further accelerating CYP discovery and engineering.
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Identification and cytochemical immunolocalization of acetyl-CoA acetyltransferase involved in the Terpenoid mevalonate pathway in Euphorbia helioscopia laticifers
Botanical Studies, 2017Co-Authors: Meng Wang, Dou Wang, Jia Chai, Qing Zhang, Yong Peng, Xia CaiAbstract:BackgroundTerpenoids, the largest class of natural products in the plant kingdom, have been widely used in medicine. The precursors of Terpenoids, isoprene phosphate (IPP) and dimethylallyl pyrophosphate (DMAPP), were synthesized from a mevalonate (MVA) pathway and a 2-C-methyl-d-erythritol-4-phosphate (MEP) pathway respectively. The acetyl-CoA acetyltransferase (AACT) is the initial enzyme in MVA pathway and is considered presently to be essential for Terpenoid backbone biosynthesis. The basic research on cytochemistry of Terpenoid metabolic enzymes is important for understanding the mechanisms underlying major metabolic processes. However, compartmentalization of AACT in plants is in controversy. Euphorbia helioscopia L. containing laticifers in the whole plant is a famous ancient folk medicine for tumor treatment, and the Terpenoid is an active ingredient. Furthermore, the laticifer cell is the main synthesizing and storing site for Terpenoids.ResultsThe gene of AACT was cloned successfully from E. helioscopia, and named as EhAACT. The EhAACT expression has no significant difference among roots, stems and leaves. However, compared with the roots and stems, the EhAACT expression level is slightly higher in leaves. In addition, EhAACT recombinant protein was expressed by procaryotic expression system and anti-EhAACT antibody was prepared, the molecular weight is about 43 kDa. Western blotting results illustrated that the EhAACT antibodies specifically recognized the endogenous proteins in E. helioscopia laticifers. At last, the subcellular localization of EhAACT in E. helioscopia laticifers was observed by using colloidal gold immune-electron microscopy. EhAACT was found to mainly distribute in the endoplasmic reticulum (ER), vacuoles originated from ER and cytosol aound vacuoles originated from ER.ConclusionsAs a result, we speculated that in E. helioscopia laticifers, EhAACT located in cytosol would be transferred to small vacuoles dilated from ER, and the precursors of Terpenoids were synthesized in these small vacuoles, then Terpenoids were further synthesized into latex particles. This result would provide theoretical basis for regulating and controlling of Terpenoid biosynthesis in laticifers.
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Identification and cytochemical immunolocalization of acetyl-CoA acetyltransferase involved in the Terpenoid mevalonate pathway in Euphorbia helioscopia laticifers.
Botanical Studies, 2017Co-Authors: Meng Wang, Dou Wang, Jia Chai, Qing Zhang, Yong PengAbstract:Terpenoids, the largest class of natural products in the plant kingdom, have been widely used in medicine. The precursors of Terpenoids, isoprene phosphate (IPP) and dimethylallyl pyrophosphate (DMAPP), were synthesized from a mevalonate (MVA) pathway and a 2-C-methyl-d-erythritol-4-phosphate (MEP) pathway respectively. The acetyl-CoA acetyltransferase (AACT) is the initial enzyme in MVA pathway and is considered presently to be essential for Terpenoid backbone biosynthesis. The basic research on cytochemistry of Terpenoid metabolic enzymes is important for understanding the mechanisms underlying major metabolic processes. However, compartmentalization of AACT in plants is in controversy. Euphorbia helioscopia L. containing laticifers in the whole plant is a famous ancient folk medicine for tumor treatment, and the Terpenoid is an active ingredient. Furthermore, the laticifer cell is the main synthesizing and storing site for Terpenoids. The gene of AACT was cloned successfully from E. helioscopia, and named as EhAACT. The EhAACT expression has no significant difference among roots, stems and leaves. However, compared with the roots and stems, the EhAACT expression level is slightly higher in leaves. In addition, EhAACT recombinant protein was expressed by procaryotic expression system and anti-EhAACT antibody was prepared, the molecular weight is about 43 kDa. Western blotting results illustrated that the EhAACT antibodies specifically recognized the endogenous proteins in E. helioscopia laticifers. At last, the subcellular localization of EhAACT in E. helioscopia laticifers was observed by using colloidal gold immune-electron microscopy. EhAACT was found to mainly distribute in the endoplasmic reticulum (ER), vacuoles originated from ER and cytosol aound vacuoles originated from ER. As a result, we speculated that in E. helioscopia laticifers, EhAACT located in cytosol would be transferred to small vacuoles dilated from ER, and the precursors of Terpenoids were synthesized in these small vacuoles, then Terpenoids were further synthesized into latex particles. This result would provide theoretical basis for regulating and controlling of Terpenoid biosynthesis in laticifers.
Michael F. Crowley - One of the best experts on this subject based on the ideXlab platform.
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Membrane Permeability of Terpenoids Explored with Molecular Simulation.
The journal of physical chemistry. B, 2018Co-Authors: Josh V. Vermaas, Gayle J. Bentley, Gregg T. Beckham, Michael F. CrowleyAbstract:Terpenoids constitute a class of compounds with remarkable potential for pharmaceutical, fragrance, specialty chemical, and biofuel applications. However, their industrial production is limited by their rarity within their native plant hosts, creating considerable interest in microbial hosts capable of manufacturing Terpenoids. To reduce production costs, nondestructive product recovery from these microbial hosts is preferred, and is achievable using a hydrophobic organic overlay. Our prior research has indicated that oxidized fatty acyl products may permeate faster through host membranes, increasing overall biorefinery productivity. To test this hypothesis for Terpenoids, we computed membrane permeabilities of conventional Terpenoid target products (e.g., limonene, bisabolene, farnesene) and related oxidized compounds through molecular dynamics simulations. These simulations indicate that Terpenoid product permeabilities from cytosol to overlay are oxidation independent, as increases in membrane extracti...
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Membrane Permeability of Terpenoids Explored with Molecular Simulation
2018Co-Authors: Josh V. Vermaas, Gayle J. Bentley, Gregg T. Beckham, Michael F. CrowleyAbstract:Terpenoids constitute a class of compounds with remarkable potential for pharmaceutical, fragrance, specialty chemical, and biofuel applications. However, their industrial production is limited by their rarity within their native plant hosts, creating considerable interest in microbial hosts capable of manufacturing Terpenoids. To reduce production costs, nondestructive product recovery from these microbial hosts is preferred, and is achievable using a hydrophobic organic overlay. Our prior research has indicated that oxidized fatty acyl products may permeate faster through host membranes, increasing overall biorefinery productivity. To test this hypothesis for Terpenoids, we computed membrane permeabilities of conventional Terpenoid target products (e.g., limonene, bisabolene, farnesene) and related oxidized compounds through molecular dynamics simulations. These simulations indicate that Terpenoid product permeabilities from cytosol to overlay are oxidation independent, as increases in membrane extraction efficiency due to product oxidation are proportionally offset by decreases in the membrane crossing rate if the membrane and organic phase are in close contact. However, if aqueous extraction is required, oxidation will accelerate the slow product extraction from the membrane. Experimental toxicity assays performed indicated that most Terpenoids tested were tolerated by microbial hosts, although exposure to oxidized terpenes often retarded microbial growth compared with conventional terpenes. Thus, Terpenoid oxidation is not expected to significantly increase or decrease the extraction productivity in an industrial setting where cells are in close contact, unlike the previously studied fatty acyl products
Jonathan Gershenzon - One of the best experts on this subject based on the ideXlab platform.
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Increased Terpenoid Accumulation in Cotton (Gossypium hirsutum) Foliage is a General Wound Response
Journal of Chemical Ecology, 2008Co-Authors: Stefan Opitz, Grit Kunert, Jonathan GershenzonAbstract:The subepidermal pigment glands of cotton accumulate a variety of Terpenoid products, including monoterpenes, sesquiterpenes, and Terpenoid aldehydes that can act as feeding deterrents against a number of insect herbivore species. We compared the effect of herbivory by Spodoptera littoralis caterpillars, mechanical damage by a fabric pattern wheel, and the application of jasmonic acid on levels of the major representatives of the three structural classes of Terpenoids in the leaf foliage of 4-week-old Gossypium hirsutum plants. Terpenoid levels increased successively from control to mechanical damage, herbivory, and jasmonic acid treatments, with E -β-ocimene and heliocide H_1 and H_4 showing the highest increases, up to 15-fold. Herbivory or mechanical damage to older leaves led to Terpenoid increases in younger leaves. Leaf-by-leaf analysis of terpenes and gland density revealed that higher levels of Terpenoids were achieved by two mechanisms: (1) increased filling of existing glands with Terpenoids and (2) the production of additional glands, which were found to be dependent on damage intensity. As the relative response of individual Terpenoids did not differ substantially among herbivore, mechanical damage, and jasmonic acid treatments, the induction of Terpenoids in cotton foliage appears to represent a non-specific wound response mediated by jasmonic acid.
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Metabolic costs of Terpenoid accumulation in higher plants
Journal of Chemical Ecology, 1994Co-Authors: Jonathan GershenzonAbstract:The net value of any plant trait can be assessed by measuring the costs and benefits associated with that trait. While the other contributors to this issue examine the possible benefits of Terpenoids to plants, this article explores the metabolic costs of Terpenoid accumulation in plants in the light of recent advances in Terpenoid biochemistry. Terpenoids are more expensive to manufacture per gram than most other primary and secondary metabolites due to their extensive chemical reduction. The enzyme costs of making Terpenoids are also high since Terpenoid biosynthetic enzymes are apparently not shared with other metabolic pathways. In fact, plant cells may even possess more than one set of enzymes for catalyzing the basic steps of Terpenoid formation. Terpenoids are usually sequestered in complex, multicellular secretory structures, and so storage costs for these substances are also likely to be substantial. However, not all of the processes involved in Terpenoid accumulation require large investments of resources. For instance, the maintenance of Terpenoid pools is probably inexpensive because there is no evidence that substantial quantities of terpenes are lost as a result of metabolic turnover, volatilization, or leaching. Moreover, plants may reduce their net Terpenoid costs by employing individual compounds in more than one role or by catabolizing substances that are no longer needed, although it is still unclear if such practices are widespread. These findings (and other facets of Terpenoid biochemistry and physiology) are discussed in relation to the assumptions and predictions of several current theories of plant defense, including the carbonnutrient balance hypothesis, the growth-differentiation balance hypothesis, and the resource availability hypothesis.
Josh V. Vermaas - One of the best experts on this subject based on the ideXlab platform.
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Membrane Permeability of Terpenoids Explored with Molecular Simulation.
The journal of physical chemistry. B, 2018Co-Authors: Josh V. Vermaas, Gayle J. Bentley, Gregg T. Beckham, Michael F. CrowleyAbstract:Terpenoids constitute a class of compounds with remarkable potential for pharmaceutical, fragrance, specialty chemical, and biofuel applications. However, their industrial production is limited by their rarity within their native plant hosts, creating considerable interest in microbial hosts capable of manufacturing Terpenoids. To reduce production costs, nondestructive product recovery from these microbial hosts is preferred, and is achievable using a hydrophobic organic overlay. Our prior research has indicated that oxidized fatty acyl products may permeate faster through host membranes, increasing overall biorefinery productivity. To test this hypothesis for Terpenoids, we computed membrane permeabilities of conventional Terpenoid target products (e.g., limonene, bisabolene, farnesene) and related oxidized compounds through molecular dynamics simulations. These simulations indicate that Terpenoid product permeabilities from cytosol to overlay are oxidation independent, as increases in membrane extracti...
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Membrane Permeability of Terpenoids Explored with Molecular Simulation
2018Co-Authors: Josh V. Vermaas, Gayle J. Bentley, Gregg T. Beckham, Michael F. CrowleyAbstract:Terpenoids constitute a class of compounds with remarkable potential for pharmaceutical, fragrance, specialty chemical, and biofuel applications. However, their industrial production is limited by their rarity within their native plant hosts, creating considerable interest in microbial hosts capable of manufacturing Terpenoids. To reduce production costs, nondestructive product recovery from these microbial hosts is preferred, and is achievable using a hydrophobic organic overlay. Our prior research has indicated that oxidized fatty acyl products may permeate faster through host membranes, increasing overall biorefinery productivity. To test this hypothesis for Terpenoids, we computed membrane permeabilities of conventional Terpenoid target products (e.g., limonene, bisabolene, farnesene) and related oxidized compounds through molecular dynamics simulations. These simulations indicate that Terpenoid product permeabilities from cytosol to overlay are oxidation independent, as increases in membrane extraction efficiency due to product oxidation are proportionally offset by decreases in the membrane crossing rate if the membrane and organic phase are in close contact. However, if aqueous extraction is required, oxidation will accelerate the slow product extraction from the membrane. Experimental toxicity assays performed indicated that most Terpenoids tested were tolerated by microbial hosts, although exposure to oxidized terpenes often retarded microbial growth compared with conventional terpenes. Thus, Terpenoid oxidation is not expected to significantly increase or decrease the extraction productivity in an industrial setting where cells are in close contact, unlike the previously studied fatty acyl products
He Huang - One of the best experts on this subject based on the ideXlab platform.
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Advances in the metabolic engineering of Yarrowia lipolytica for the production of Terpenoids.
Bioresource technology, 2019Co-Authors: Ma Yirong, Kai-feng Wang, Wei-jian Wang, Ying Ding, Tian-qiong Shi, He HuangAbstract:Terpenoids are a large class of natural compounds based on the C5 isoprene unit, with many biological effects such activity against cancer and allergies, while some also have an agreeable aroma. Consequently, they have received extensive attention in the food, pharmaceutical and cosmetic fields. With the identification and analysis of the underlying natural product synthesis pathways, current microbial-based metabolic engineering approaches have yielded new strategies for the production of highly valuable Terpenoids. Yarrowia lipolytica is a non-conventional oleaginous yeast that is rapidly emerging as a valuable host for the production of Terpenoids due to its own endogenous mevalonate pathway and high oil production capacity. This review aims to summarize the status and strategies of metabolic engineering for the heterologous synthesis of Terpenoids in Y. lipolytica in recent years and proposes new methods aiming towards further improvement of Terpenoid production.
