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      Natural rubber biosynthesis in plants, the rubber transferase complex, and metabolic engineering progress and prospects

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          Summary

          Natural rubber ( NR) is a nonfungible and valuable biopolymer, used to manufacture ~50 000 rubber products, including tires and medical gloves. Current production of NR is derived entirely from the para rubber tree ( Hevea brasiliensis). The increasing demand for NR, coupled with limitations and vulnerability of H. brasiliensis production systems, has induced increasing interest among scientists and companies in potential alternative NR crops. Genetic/metabolic pathway engineering approaches, to generate NR‐enriched genotypes of alternative NR plants, are of great importance. However, although our knowledge of rubber biochemistry has significantly advanced, our current understanding of NR biosynthesis, the biosynthetic machinery and the molecular mechanisms involved remains incomplete. Two spatially separated metabolic pathways provide precursors for NR biosynthesis in plants and their genes and enzymes/complexes are quite well understood. In contrast, understanding of the proteins and genes involved in the final step(s)—the synthesis of the high molecular weight rubber polymer itself—is only now beginning to emerge. In this review, we provide a critical evaluation of recent research developments in NR biosynthesis, in vitro reconstitution, and the genetic and metabolic pathway engineering advances intended to improve NR content in plants, including H. brasiliensis, two other prospective alternative rubber crops, namely the rubber dandelion and guayule, and model species, such as lettuce. We describe a new model of the rubber transferase complex, which integrates these developments. In addition, we highlight the current challenges in NR biosynthesis research and future perspectives on metabolic pathway engineering of NR to speed alternative rubber crop commercial development.

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          Most cited references218

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          Semi-synthetic artemisinin: a model for the use of synthetic biology in pharmaceutical development.

          Recent developments in synthetic biology, combined with continued progress in systems biology and metabolic engineering, have enabled the engineering of microorganisms to produce heterologous molecules in a manner that was previously unfeasible. The successful synthesis and recent entry of semi-synthetic artemisinin into commercial production is the first demonstration of the potential of synthetic biology for the development and production of pharmaceutical agents. In this Review, we describe the metabolic engineering and synthetic biology approaches that were used to develop this important antimalarial drug precursor. This not only demonstrates the incredible potential of the available technologies but also illuminates how lessons learned from this work could be applied to the production of other pharmaceutical agents.
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            Crosstalk between cytosolic and plastidial pathways of isoprenoid biosynthesis in Arabidopsis thaliana.

            In plants, the formation of isopentenyl diphosphate and dimethylallyl diphosphate, the central intermediates in the biosynthesis of isoprenoids, is compartmentalized: the mevalonate (MVA) pathway, which is localized to the cytosol, is responsible for the synthesis of sterols, certain sesquiterpenes, and the side chain of ubiquinone; in contrast, the recently discovered MVA-independent pathway, which operates in plastids, is involved in providing the precursors for monoterpenes, certain sesquiterpenes, diterpenes, carotenoids, and the side chains of chlorophylls and plastoquinone. Specific inhibitors of the MVA pathway (lovastatin) and the MVA-independent pathway (fosmidomycin) were used to perturb biosynthetic flux in Arabidopsis thaliana seedlings. The interaction between both pathways was studied at the transcriptional level by using GeneChip (Affymetrix) microarrays and at the metabolite level by assaying chlorophylls, carotenoids, and sterols. Treatment of seedlings with lovastatin resulted in a transient decrease in sterol levels and a transient increase in carotenoid as well as chlorophyll levels. After the initial drop, sterol amounts in lovastatin-treated seedlings recovered to levels above controls. As a response to fosmidomycin treatment, a transient increase in sterol levels was observed, whereas chlorophyll and carotenoid amounts decreased dramatically when compared with controls. At 96 h after fosmidomycin addition, the levels of all metabolites assayed (sterols, chlorophylls, and carotenoids) were substantially lower than in controls. Interestingly, these inhibitor-mediated changes were not reflected in altered gene expression levels of the genes involved in sterol, chlorophyll, and carotenoid metabolism. The lack of correlation between gene expression patterns and the accumulation of isoprenoid metabolites indicates that posttranscriptional processes may play an important role in regulating flux through isoprenoid metabolic pathways.
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              Regulation of sterol synthesis in eukaryotes.

              Cholesterol is an essential component of mammalian cell membranes and is required for proper membrane permeability, fluidity, organelle identity, and protein function. Cells maintain sterol homeostasis by multiple feedback controls that act through transcriptional and posttranscriptional mechanisms. The membrane-bound transcription factor sterol regulatory element binding protein (SREBP) is the principal regulator of both sterol synthesis and uptake. In mammalian cells, the ER membrane protein Insig has emerged as a key component of homeostatic regulation by controlling both the activity of SREBP and the sterol-dependent degradation of the biosynthetic enzyme HMG-CoA reductase. In this review, we focus on recent advances in our understanding of the molecular mechanisms of the regulation of sterol synthesis. A comparative analysis of SREBP and HMG-CoA reductase regulation in mammals, yeast, and flies points toward an equilibrium model for how lipid signals regulate the activity of sterol-sensing proteins and their downstream effectors.
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                Author and article information

                Contributors
                sbryu@kribb.re.kr
                cornish.19@osu.edu
                Journal
                Plant Biotechnol J
                Plant Biotechnol. J
                10.1111/(ISSN)1467-7652
                PBI
                Plant Biotechnology Journal
                John Wiley and Sons Inc. (Hoboken )
                1467-7644
                1467-7652
                26 June 2019
                November 2019
                : 17
                : 11 ( doiID: 10.1111/pbi.v17.11 )
                : 2041-2061
                Affiliations
                [ 1 ] Plant Systems Engineering Research Centre Korea Research Institute of Bioscience and Biotechnology (KRIBB) Yuseong‐gu Daejeon Korea
                [ 2 ] Research & Development Center DRB Holding Co. LTD Busan Korea
                [ 3 ] Department of Biosystems and Bioengineering KRIBB School of Biotechnology Korea University of Science and Technology (UST) Daejeon Korea
                [ 4 ] Department of Horticulture and Crop Science The Ohio State University Wooster OH USA
                [ 5 ] Department of Food, Agricultural and Biological Engineering The Ohio State University Wooster OH USA
                Author notes
                [*] [* ] Correspondence (Tel +1 330 263 3982; fax +1 330 263 3777 (K.C.); emails cornish.19@ 123456osu.edu (K.C.) and Tel +82 42 8604295; fax +82 42 8604489 (S.B.R.); sbryu@ 123456kribb.re.kr (S.B.R.))
                Author information
                https://orcid.org/0000-0002-3327-2454
                Article
                PBI13181
                10.1111/pbi.13181
                6790360
                31150158
                0ecc648c-ddeb-45ad-823f-a682027ad879
                © 2019 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

                History
                : 03 December 2018
                : 24 May 2019
                : 29 May 2019
                Page count
                Figures: 3, Tables: 2, Pages: 21, Words: 20641
                Funding
                Funded by: United States Department of Agriculture, National Institute of Food and Agriculture , open-funder-registry 10.13039/100000199;
                Award ID: 230837
                Funded by: DRB Holding Co. LTD
                Funded by: Brain Pool program of Korean government
                Funded by: Cooperative Research Program for Agriculture Science & Technology Development
                Award ID: PJ01368201 & PJ013486
                Categories
                Review
                Reviews
                Custom metadata
                2.0
                November 2019
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.7.0 mode:remove_FC converted:14.10.2019

                Biotechnology
                allylic pyrophosphate,cis‐prenyl transferase,guayule,hevea brasiliensis,isopentenyl pyrophosphate,natural rubber,parthenium argentatum,rubber dandelion,rubber transferase,taraxacum kok‐saghyz

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