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Predictive Engineering of Class I Terpene Synthases Using Experimental and Computational Approaches.Chembiochem : a European Journal of... Mar 2022Terpenoids are a highly diverse group of natural products with considerable industrial interest. Increasingly, engineered microbes are used for the production of... (Review)
Review
Terpenoids are a highly diverse group of natural products with considerable industrial interest. Increasingly, engineered microbes are used for the production of terpenoids to replace natural extracts and chemical synthesis. Terpene synthases (TSs) show a high level of functional plasticity and are responsible for the vast structural diversity observed in natural terpenoids. Their relatively inert active sites guide intrinsically reactive linear carbocation intermediates along one of many cyclisation paths via exertion of subtle steric and electrostatic control. Due to the absence of a strong protein interaction with these intermediates, there is a remarkable lack of sequence-function relationship within the TS family, making product-outcome predictions from sequences alone challenging. This, in combination with the fact that many TSs produce multiple products from a single substrate hampers the design and use of TSs in the biomanufacturing of terpenoids. This review highlights recent advances in genome mining, computational modelling, high-throughput screening, and machine-learning that will allow more predictive engineering of these fascinating enzymes in the near future.
Topics: Alkyl and Aryl Transferases; Catalytic Domain; Cyclization; Terpenes
PubMed: 34669250
DOI: 10.1002/cbic.202100484 -
Bioscience, Biotechnology, and... 2013To the best of my knowledge, only 19 cyclic and 8 linear C35 terpenes have been identified to date, and no family name was assigned to this terpene class until recently.... (Review)
Review
To the best of my knowledge, only 19 cyclic and 8 linear C35 terpenes have been identified to date, and no family name was assigned to this terpene class until recently. In 2011, it was proposed that these C35 terpenes should be called sesquarterpenes. This review highlights the biosynthesis of two kinds of sesquarterpenes (C35 terpenes) that are produced via cyclization of a linear C35 isoprenoid in Bacillus and Mycobacterium species. In Bacillus species, a new type of terpene cyclase that has no sequence homology with any known terpene synthases, as well as a bifunctional terpene cyclase that biosynthesizes two classes of cyclic terpenes with different numbers of carbons as natural products, have been identified. On the other hand, in Mycobacterium species, the first bifunctional Z-prenyltransferase has been found, but a novel terpene cyclase and a unique polyprenyl reductase remain unidentified. The identification of novel enzyme types should lead to the discovery of many homologous enzymes and their products including novel natural compounds. On the other hand, many enzymes responsible for the biosynthesis of natural products have low substrate specificities in vitro. Therefore, to find novel natural products present in organisms, the multifunctionality of enzymes in the biosynthetic pathway of natural products should be analyzed.
Topics: Alkyl and Aryl Transferases; Bacillus; Cyclization; Dimethylallyltranstransferase; Mycobacterium; Substrate Specificity; Terpenes
PubMed: 23748782
DOI: 10.1271/bbb.130180 -
The Plant Journal : For Cell and... Nov 2021Plants interact with other organisms employing volatile organic compounds (VOCs). The largest group of plant-released VOCs are terpenes, comprised of isoprene,... (Review)
Review
Plants interact with other organisms employing volatile organic compounds (VOCs). The largest group of plant-released VOCs are terpenes, comprised of isoprene, monoterpenes, and sesquiterpenes. Mono- and sesquiterpenes are well-known communication compounds in plant-insect interactions, whereas the smallest, most commonly emitted terpene, isoprene, is rather assigned a function in combating abiotic stresses. Recently, it has become evident that different volatile terpenes also act as plant-to-plant signaling cues. Upon being perceived, specific volatile terpenes can sensitize distinct signaling pathways in receiver plant cells, which in turn trigger plant innate immune responses. This vastly extends the range of action of volatile terpenes, which not only protect plants from various biotic and abiotic stresses, but also convey information about environmental constraints within and between plants. As a result, plant-insect and plant-pathogen interactions, which are believed to influence each other through phytohormone crosstalk, are likely equally sensitive to reciprocal regulation via volatile terpene cues. Here, we review the current knowledge of terpenes as volatile semiochemicals and discuss why and how volatile terpenes make good signaling cues. We discuss how volatile terpenes may be perceived by plants, what are possible downstream signaling events in receiver plants, and how responses to different terpene cues might interact to orchestrate the net plant response to multiple stresses. Finally, we discuss how the signal can be further transmitted to the community level leading to a mutually beneficial community-scale response or distinct signaling with near kin.
Topics: Plant Cells; Plant Growth Regulators; Plant Immunity; Plants; Signal Transduction; Species Specificity; Terpenes; Volatile Organic Compounds
PubMed: 34369010
DOI: 10.1111/tpj.15453 -
Plant Biotechnology Journal Feb 2015Photosynthetic hydrocarbon production bypasses the traditional biomass hydrolysis process and represents the most direct conversion of sunlight energy into the... (Review)
Review
Photosynthetic hydrocarbon production bypasses the traditional biomass hydrolysis process and represents the most direct conversion of sunlight energy into the next-generation biofuels. As a major class of biologically derived hydrocarbons with diverse structures, terpenes are also valuable in producing a variety of fungible bioproducts in addition to the advanced 'drop-in' biofuels. However, it is highly challenging to achieve the efficient redirection of photosynthetic carbon and reductant into terpene biosynthesis. In this review, we discuss four major scientific and technical barriers for photosynthetic terpene production and recent advances to address these constraints. Collectively, photosynthetic terpene production needs to be optimized in a systematic fashion, in which the photosynthesis improvement, the optimization of terpene biosynthesis pathway, the improvement of key enzymes and the enhancement of sink effect through terpene storage or secretion are all important. New advances in synthetic biology also offer a suite of potential tools to design and engineer photosynthetic terpene platforms. The systemic integration of these solutions may lead to 'disruptive' technologies to enable biofuels and bioproducts with high efficiency, yield and infrastructure compatibility.
Topics: Biofuels; Biosynthetic Pathways; Photosynthesis; Phytochemicals; Terpenes
PubMed: 25626473
DOI: 10.1111/pbi.12343 -
Current Opinion in Biotechnology Jun 2017Advances in retooling microorganisms have enabled bioproduction of 'drop-in' biofuels, fuels that are compatible with existing spark-ignition, compression-ignition, and... (Review)
Review
Advances in retooling microorganisms have enabled bioproduction of 'drop-in' biofuels, fuels that are compatible with existing spark-ignition, compression-ignition, and gas-turbine engines. As the majority of petroleum consumption in the United States consists of gasoline (47%), diesel fuel and heating oil (21%), and jet fuel (8%), 'drop-in' biofuels that replace these petrochemical sources are particularly attractive. In this review, we discuss the application of aldehyde decarbonylases to produce gasoline substitutes from fatty acid products, a recently crystallized reductase that could hydrogenate jet fuel precursors from terpene synthases, and the exquisite control of polyketide synthases to produce biofuels with desired physical properties (e.g., lower freezing points). With our increased understanding of biosynthetic logic of metabolic pathways, we discuss the unique advantages of fatty acid, terpene, and polyketide synthases for the production of bio-based gasoline, diesel and jet fuel.
Topics: Bacteria; Biofuels; Biosynthetic Pathways; Gasoline; Hydrocarbons; Petroleum; Polyketide Synthases; Terpenes
PubMed: 28427010
DOI: 10.1016/j.copbio.2017.03.004 -
International Journal of Molecular... Jun 2022Dendrobium catenatum is a widely cultivated Chinese orchid herb rich in abundant secondary metabolites, such as terpenes. However, terpene distribution and...
Dendrobium catenatum is a widely cultivated Chinese orchid herb rich in abundant secondary metabolites, such as terpenes. However, terpene distribution and characterization of terpene biosynthesis-related genes remain unknown in D. catenatum. In this study, metabolic profiling was performed to analyze terpene distribution in the root, stem, leaf, and flower of D. catenatum. A total of 74 terpene compounds were identified and classified. Clustering analysis revealed that terpene compounds exhibited a tissue-specific accumulation, including monoterpenes in the flowers, sesquiterpenes in the stems, and triterpenes in the roots. Transcriptome analysis revealed that the ‘terpenoid backbone biosynthesis’ pathway was only significantly enriched in root vs. flower. The expression of terpene biosynthesis-related genes was spatiotemporal in the flowers. Prenylsynthase-terpene synthases (PS-TPSs) are the largest and core enzymes for generating terpene diversity. By systematic sequence analysis of six species, 318 PS-TPSs were classified into 10 groups and 51 DcaPS-TPSs were found in eight of them. Eighteen DcaPS-TPSs were regulated by circadian rhythm under drought stress. Most of the DcaPS-TPSs were influenced by cold stress and fungi infection. The cis-element of the majority of the DcaPS-TPS promoters was related to abiotic stress and plant development. Methyl jasmonate levels were significantly associated with DcaTPSs expression and terpene biosynthesis. These results provide insight into further functional investigation of DcaPS-TPSs and the regulation of terpene biosynthesis in Dendrobium.
Topics: Alkyl and Aryl Transferases; Dendrobium; Gene Expression Regulation, Plant; Phylogeny; Plant Proteins; Stress, Physiological; Terpenes
PubMed: 35742843
DOI: 10.3390/ijms23126398 -
Plant Cell Reports Sep 2022Terpenoids are synthesized naturally by plants as secondary metabolites, and are diverse and complex in structure with multiple applications in bioenergy, food,... (Review)
Review
Terpenoids are synthesized naturally by plants as secondary metabolites, and are diverse and complex in structure with multiple applications in bioenergy, food, cosmetics, and medicine. This makes the production of terpenoids such as isoprene, β-phellandrene, farnesene, amorphadiene, and squalene valuable, owing to which their industrial demand cannot be fulfilled exclusively by plant sources. They are synthesized via the Methylerythritol phosphate pathway (MEP) and the Mevalonate pathway (MVA), both existing in plants. The advent of genetic engineering and the latest accomplishments in synthetic biology and metabolic engineering allow microbial synthesis of terpenoids. Cyanobacteria manifest to be the promising hosts for this, utilizing sunlight and CO. Cyanobacteria possess MEP pathway to generate precursors for terpenoid synthesis. The terpenoid synthesis can be amplified by overexpressing the MEP pathway and engineering MVA pathway genes. According to the desired terpenoid, terpene synthases unique to the plant kingdom must be incorporated in cyanobacteria. Engineering an organism to be used as a cell factory comes with drawbacks such as hampered cell growth and disturbance in metabolic flux. This review set forth a comparison between MEP and MVA pathways, strategies to overexpress these pathways with their challenges.
Topics: Cyanobacteria; Metabolic Engineering; Mevalonic Acid; Plants; Terpenes
PubMed: 35789422
DOI: 10.1007/s00299-022-02892-9 -
Molecules (Basel, Switzerland) Jan 2023Interest in cultivating cannabis for medical and recreational purposes is increasing due to a dramatic shift in cannabis legislation worldwide. Therefore, a...
Interest in cultivating cannabis for medical and recreational purposes is increasing due to a dramatic shift in cannabis legislation worldwide. Therefore, a comprehensive understanding of the composition of secondary metabolites, cannabinoids, and terpenes grown in different environmental conditions is of primary importance for the medical and recreational use of cannabis. We compared the terpene and cannabinoid profiles using gas/liquid chromatography and mass spectrometry for commercial cannabis from genetically identical plants grown indoors using artificial light and artificially grown media or outdoors grown in living soil and natural sunlight. By analyzing the cannabinoids, we found significant variations in the metabolomic profile of cannabis for the different environments. Overall, for both cultivars, there were significantly greater oxidized and degraded cannabinoids in the indoor-grown samples. Moreover, the outdoor-grown samples had significantly more unusual cannabinoids, such as C4- and C6-THCA. There were also significant differences in the terpene profiles between indoor- and outdoor-grown cannabis. The outdoor samples had a greater preponderance of sesquiterpenes including β-caryophyllene, α-humulene, α-bergamotene, α-guaiene, and germacrene B relative to the indoor samples.
Topics: Cannabinoids; Cannabis; Terpenes; Gas Chromatography-Mass Spectrometry; Hallucinogens; Cannabinoid Receptor Agonists
PubMed: 36677891
DOI: 10.3390/molecules28020833 -
PloS One 2017Cannabis (Cannabis sativa) plants produce and accumulate a terpene-rich resin in glandular trichomes, which are abundant on the surface of the female inflorescence....
Cannabis (Cannabis sativa) plants produce and accumulate a terpene-rich resin in glandular trichomes, which are abundant on the surface of the female inflorescence. Bouquets of different monoterpenes and sesquiterpenes are important components of cannabis resin as they define some of the unique organoleptic properties and may also influence medicinal qualities of different cannabis strains and varieties. Transcriptome analysis of trichomes of the cannabis hemp variety 'Finola' revealed sequences of all stages of terpene biosynthesis. Nine cannabis terpene synthases (CsTPS) were identified in subfamilies TPS-a and TPS-b. Functional characterization identified mono- and sesqui-TPS, whose products collectively comprise most of the terpenes of 'Finola' resin, including major compounds such as β-myrcene, (E)-β-ocimene, (-)-limonene, (+)-α-pinene, β-caryophyllene, and α-humulene. Transcripts associated with terpene biosynthesis are highly expressed in trichomes compared to non-resin producing tissues. Knowledge of the CsTPS gene family may offer opportunities for selection and improvement of terpene profiles of interest in different cannabis strains and varieties.
Topics: Acyclic Monoterpenes; Alkenes; Alkyl and Aryl Transferases; Bicyclic Monoterpenes; Cannabis; Cyclohexenes; Gene Expression Regulation, Plant; Inflorescence; Isoenzymes; Limonene; Metabolic Networks and Pathways; Monocyclic Sesquiterpenes; Monoterpenes; Multigene Family; Phylogeny; Polycyclic Sesquiterpenes; RNA, Messenger; Sesquiterpenes; Terpenes
PubMed: 28355238
DOI: 10.1371/journal.pone.0173911 -
BMC Complementary and Alternative... Jun 2017Volatile oil from aromatic plants has been used by ancient Egyptians in embalming for the inhibition of bacterial growth and prevention of decay, Callistemon citrinus is...
Terpene constituents of the aerial parts, phenolic content, antibacterial potential, free radical scavenging and antioxidant activity of Callistemon citrinus (Curtis) Skeels (Myrtaceae) from Eastern Cape Province of South Africa.
BACKGROUND
Volatile oil from aromatic plants has been used by ancient Egyptians in embalming for the inhibition of bacterial growth and prevention of decay, Callistemon citrinus is used in traditional therapies for the treatment of bronchitis, cough, inflammation and as an antimicrobial herbs. This study examines the essential constituents of the volatile oils obtained from the aerial parts of the plant as well as its antioxidant activity, free radical scavenging, phenolic content and the antibacterial potential of the oils.
METHODS
A portion of 500 g, 250 g and 150 g of the leaves, flowers and stems of this plant respectively were subjected to hydro-distillation process for three hours. The oils collected from the various plant parts were immediately subjected to GC-MS analysis. The overall phenolic content of the leaves oil, radical scavenging, antibacterial action and antioxidant activities of the essential oils of both the leaves and flowers of Callistemon citrinus were determined using standard methods, with free radical DPPH and ABTS as a reference antioxidant.
RESULTS
Analyses of the three oils revealed a total of twenty-six components for the leaves oil representing 96.84% of the total oil composition, forty-one components for the flowers oil accounting for 98.92% of the whole composition and ten components for the stem oil amounting to 99.98% of the entire oil constituents. The dominant compounds in the leaves oil were eucalyptol (48.98%) and α-terpineol (8.01%), while α-eudesmol (12.93%), caryophyllene (11.89%), (-)-bornyl-acetate (10.02%) and eucalyptol (8.11%) were the main constituents of the flowers oil. In the same vein, the leading constituents in the stems oil were eucalyptol (56.00%) and α-pinene (31.03%). The antioxidant capacities of both the leaves and flowers oils of the plant were evaluated and their IC were (1.49 and 1.13) for DPPH and (0.14 and 0.03) for ABTS assay respectively. The antibacterial activities of the oils from the (leaves and flowers) were also examined and were found to have wide range of activities against the bacterial strains used in this study.
CONCLUSION
Observations drawn from this experiment shows clearly that the leaves and flowers of Callistemon citrinus possess phenolic compounds and cyclic ether of several pharmacological behaviors.
Topics: Anti-Bacterial Agents; Antioxidants; Bacteria; Flowers; Gas Chromatography-Mass Spectrometry; Myrtaceae; Plant Extracts; Plant Leaves; Plant Stems; South Africa; Terpenes
PubMed: 28583128
DOI: 10.1186/s12906-017-1804-2