Green Way of Biomedicine – How to Force Plants to Produce New Important Proteins

Wiktorek-Smagur, Aneta; Hnatuszko-Konka, Katarzyna; Gerszberg, Aneta; Kowalczyk, Tomasz; Luchniak, Piotr; Andrzej, K.

doi:10.5772/31145

Cited by 6 publications

(14 citation statements)

References 37 publications

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“…Recombinant proteins are currently being produced in cultured cell-based systems in mammals, microbes (bacteria and yeast), insects and plants, as well as in transgenic animals (reviewed by Demain and Vaishnav) 1 . Transgenic plants constitute an attractive system for expression and production of a variety of proteins and biomolecules due to their efficient eukaryotic protein synthesis, high scalability, relatively low production costs and environmental footprint 2 – 4 . However, selecting suitable hosts and expression vectors are key considerations since protein accumulation is determined by expression levels.…”

Section: Introductionmentioning

confidence: 99%

Unprecedented enhancement of recombinant protein production in sugarcane culms using a combinatorial promoter stacking system

Damaj¹,

Jifon

Woodard

et al. 2020

Sci Rep

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Plants represent a safe and cost-effective platform for producing high-value proteins with pharmaceutical properties; however, the ability to accumulate these in commercially viable quantities is challenging. Ideal crops to serve as biofactories would include low-input, fast-growing, highbiomass species such as sugarcane. The objective of this study was to develop an efficient expression system to enable large-scale production of high-value recombinant proteins in sugarcane culms. Bovine lysozyme (BvLz) is a potent broad-spectrum antimicrobial enzyme used in the food, cosmetics and agricultural industries. Here, we report a novel strategy to achieve high-level expression of recombinant proteins using a combinatorial stacked promoter system. We demonstrate this by co-expressing BvLz under the control of multiple constitutive and culm-regulated promoters on separate expression vectors and combinatorial plant transformation. BvLz accumulation reached 1.4% of total soluble protein (TSP) (10.0 mg BvLz/kg culm mass) in stacked multiple promoter:BvLz lines, compared to 0.07% of TSP (0.56 mg/kg) in single promoter:BvLz lines. BvLz accumulation was further boosted to 11.5% of TSP (82.5 mg/kg) through event stacking by re-transforming the stacked promoter:BvLz lines with additional BvLz expression vectors. The protein accumulation achieved with the combinatorial promoter stacking expression system was stable in multiple vegetative propagations, demonstrating the feasibility of using sugarcane as a biofactory for producing highvalue proteins and bioproducts. Recombinant proteins are currently being produced in cultured cell-based systems in mammals, microbes (bacteria and yeast), insects and plants, as well as in transgenic animals (reviewed by Demain and Vaishnav) 1. Transgenic plants constitute an attractive system for expression and production of a variety of proteins and biomolecules due to their efficient eukaryotic protein synthesis, high scalability, relatively low production costs and environmental footprint 2-4. However, selecting suitable hosts and expression vectors are key considerations since protein accumulation is determined by expression levels. Important factors to consider when selecting a plant-based production platform include biomass yield per hectare, recombinant protein yield per unit biomass, ease of transformation, scalability and safety 5. Sugarcane

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Section: Introductionmentioning

confidence: 99%

Unprecedented enhancement of recombinant protein production in sugarcane culms using a combinatorial promoter stacking system

Damaj¹,

Jifon

Woodard

et al. 2020

Sci Rep

View full text Add to dashboard Cite

show abstract

“…A number of studies over the past decades have proved that transgenic plants (including tomatoes) can be used as bioreactors for the production of recombinant therapeutic proteins (Wiktorek-Smagur et al 2012). Zhang et al (2007), using Agrobacterium-mediated transformation, demonstrated that the hFIX (human coagulation Factor IX) gene was expressed specifically in tomato fruits.…”

Section: Molecular Farming Of Tomatoesmentioning

confidence: 99%

“…edible vaccines), the purification process can be omitted. Whereas transgenic plants or virus-infected plants can be grown on field requiring only water, minerals and sunlight, mammalian cell cultivation is a very expensive process, requiring bioreactors that cost several hundred million dollars when production is scaled up to commercial levels So far, more than 100 diagnostic and therapeutic recombinant proteins as well as vaccines have been produced in various plants including tobacco, potato, tomato, lettuce, carrot, cereals, legumes (Wiktorek-Smagur et al 2012). Here, we present some examples of the production of recombinant pharmaceutical proteins in tomatoes.…”

Section: Molecular Farming Of Tomatoesmentioning

confidence: 99%

Tomato (Solanum lycopersicum L.) in the service of biotechnology

Gerszberg

Hnatuszko-Konka

Kowalczyk

et al. 2014

Plant Cell Tiss Organ Cult

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152

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Originating in the Andes, the tomato (Solanum lycopersicum L.) was imported to Europe in the 16th century. At present, it is an important crop plant cultivated all over the world, and its production and consumption continue to increase. This popular vegetable is known as a major source of important nutrients including lycopene, bcarotene, flavonoids and vitamin C as well as hydroxycinnamic acid derivatives. Since the discovery that lycopene has anti-oxidative, anti-cancer properties, interest in tomatoes has grown rapidly. The development of genetic engineering tools and plant biotechnology has opened great opportunities for engineering tomato plants. This review presents examples of successful tissue culture and genetically modified tomatoes which resistance to a range of environmental stresses improved, along with fruit quality. Additionally, a successful molecular farming model was established.

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“…To treat these protein deficiencies the missing or dysfunctional molecules are complemented or substituted with therapeutics provided by different biological systems. However, protein therapeutics must unavoidably adhere to quality constraints that are much stricter than those for chemical industries [1, 2]. Although it is undoubtedly a challenging task to obtain an active protein in a way that is economically feasible, biopharmaceuticals (recombinant proteins, monoclonal antibodies, or vaccines) are the largest group of drugs developed in the pharmaceutical industry [3].…”

Section: Introductionmentioning

confidence: 99%

A Brief Reminder of Systems of Production and Chromatography-Based Recovery of Recombinant Protein Biopharmaceuticals

Owczarek

Gerszberg

Hnatuszko-Konka

2019

BioMed Research International

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Recombinant proteins are produced for various applications in laboratory and industrial settings. Among them, therapeutic applications have evolved into a mature field in recent years, affecting the face of contemporary medical treatment. This, in turn, has stimulated an ever-greater need for innovative technologies for the description, expression, and purification of recombinant protein biopharmaceuticals. Therefore, many biopharmaceuticals are synthesized in heterologous systems to obtain satisfactory yields that cannot be provided by natural sources. As more than 35 years has passed since the first recombinant biopharmaceutical (human insulin) successfully completed clinical trials in humans, we provide a brief review of the available prokaryotic and eukaryotic expression systems, listing the advantages and disadvantages of their use. Some examples of therapeutic proteins expressed in heterologous hosts are also provided. Moreover, technologies for the universal extraction of protein molecules are mentioned here, as is the methodology of their purification.

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Green Way of Biomedicine – How to Force Plants to Produce New Important Proteins

Cited by 6 publications

References 37 publications

Unprecedented enhancement of recombinant protein production in sugarcane culms using a combinatorial promoter stacking system

Unprecedented enhancement of recombinant protein production in sugarcane culms using a combinatorial promoter stacking system

Tomato (Solanum lycopersicum L.) in the service of biotechnology

A Brief Reminder of Systems of Production and Chromatography-Based Recovery of Recombinant Protein Biopharmaceuticals

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