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      Plant based production of myoglobin - a novel source of the muscle heme-protein

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          Abstract

          Myoglobin is a heme-protein in the muscle of vertebrates with important functions in the oxygenation of tissues and as a regulator in nitric oxide signaling. Myoglobin from many species is also an important nutritional source of bioavailable iron. In this study, we have successfully produced human myoglobin in the leaves of Nicotiana benthamiana by transient expression using a viral vector delivered by Agrobacterium tumefaciens. Analyses confirmed that heme was incorporated and the protein was functional, with observed properties consistent with those of native myoglobins. A relatively high degree of purity could be achieved with low cost methods. The results show the high potential of plants as a production platform for heme proteins, a group of proteins of interest for iron nutrition applications and possible future pharmaceutical development.

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          Systemic Agrobacterium tumefaciens-mediated transfection of viral replicons for efficient transient expression in plants.

          Plant biotechnology relies on two approaches for delivery and expression of heterologous genes in plants: stable genetic transformation and transient expression using viral vectors. Although much faster, the transient route is limited by low infectivity of viral vectors carrying average-sized or large genes. We have developed constructs for the efficient delivery of RNA viral vectors as DNA precursors and show here that Agrobacterium-mediated delivery of these constructs results in gene amplification in all mature leaves of a plant simultaneously (systemic transfection). This process, called "magnifection", can be performed on a large scale and with different plant species. This technology combines advantages of three biological systems (the transfection efficiency of A. tumefaciens, the high expression yield obtained with viral vectors, and the post-translational capabilities of a plant), does not require genetic modification of plants and is faster than other existing methods.
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            Structure of Myoglobin: A Three-Dimensional Fourier Synthesis at 2 Å. Resolution

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              TRBO: a high-efficiency tobacco mosaic virus RNA-based overexpression vector.

              Transient expression is a rapid, useful approach for producing proteins of interest in plants. Tobacco mosaic virus (TMV)-based transient expression vectors can express very high levels of foreign proteins in plants. However, TMV vectors are, in general, not efficiently delivered to plant cells by agroinfection. It was determined that agroinfection was very efficient with a 35S promoter-driven TMV replicon that lacked the TMV coat protein gene sequence. This coat protein deletion vector had several useful features as a transient expression system, including improved ease of use, higher protein expression rates, and improved biocontainment. Using this TMV expression vector, some foreign proteins were expressed at levels of 3 to 5 mg/g fresh weight of plant tissue. It is proposed that this new transient expression vector will be a useful tool for expressing recombinant proteins in plants for either research or production purposes.
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                Author and article information

                Contributors
                Li-Hua.Zhu@slu.se
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                22 January 2020
                22 January 2020
                2020
                : 10
                : 920
                Affiliations
                [1 ]ISNI 0000 0000 8578 2742, GRID grid.6341.0, Department of Plant Breeding, , Swedish University of Agricultural Sciences, ; Box 101, 230 53 Alnarp, Sweden
                [2 ]ISNI 0000 0001 0930 2361, GRID grid.4514.4, Division of Pure and Applied Biochemistry, Department of Chemistry, , Lund University, ; Box 124, 221 00 Lund, Sweden
                Article
                57565
                10.1038/s41598-020-57565-y
                6976567
                31969582
                6c6a5951-1b21-45be-90e7-c1dcde7ef1f2
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 19 July 2019
                : 16 December 2019
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                © The Author(s) 2020

                Uncategorized
                plant biotechnology,biotechnology,molecular biology,plant sciences
                Uncategorized
                plant biotechnology, biotechnology, molecular biology, plant sciences

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