Unprecedented enhancement of recombinant protein production in sugarcane culms using a combinatorial promoter stacking system
| Autor: | Kranthi K. Mandadi, Zivko L. Nikolov, Joe Molina, Carol Vargas-Bautista, Bassam B. Damaj, John L. Jifon, Georgia O. F. Barros, Steven G. White, Mona B. Damaj, Susan L. Woodard |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Biomaterials - proteins Expression systems Stacking lcsh:Medicine Article law.invention 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Transformation Genetic law Bioproducts Animals lcsh:Science Promoter Regions Genetic chemistry.chemical_classification Multidisciplinary Expression vector lcsh:R Promoter Plants Genetically Modified Recombinant Proteins Saccharum Transformation (genetics) 030104 developmental biology Enzyme chemistry Biochemistry Biofuels Recombinant DNA lcsh:Q Cattle Muramidase Lysozyme Plant sciences 030217 neurology & neurosurgery |
| Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-16 (2020) |
| ISSN: | 2045-2322 |
| Popis: | 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, high-biomass 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 high-value proteins and bioproducts. |
| Databáze: | OpenAIRE |
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