ArticleBiotechnology for biofuels2013
Optimization of transplastomic production of hemicellulases in tobacco: effects of expression cassette configuration and tobacco cultivar used as production platform on recombinant protein yields.
Article in Biotechnology for biofuels, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 46 citations in OpenAlex.
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- Plastid Transformation: New Challenges in the Circular Economy Era.International journal of molecular sciences · 2022Review
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- Improvement of recombinant miraculin production in transgenic tomato by crossbreeding-based genetic background modification.Transgenic research · 2022Article
- Mitigation of deleterious phenotypes in chloroplast-engineered plants accumulating high levels of foreign proteins.Biotechnology for biofuels · 2021Article
- Antibodies induced by oral immunization of mice with a recombinant protein produced in tobacco plants harboringPlant cell, tissue and organ culture · 2021Article
- Plastid Transformation: How Does it Work? Can it Be Applied to Crops? What Can it Offer?International journal of molecular sciences · 2020Review
- RNA Interference in the Tobacco Hornworm,Frontiers in plant science · 2019Article
- Co-expression with the Type 3 Secretion Chaperone CesT from EnterohemorrhagicFrontiers in plant science · 2017Article
- Strategies for the production of cell wall-deconstructing enzymes in lignocellulosic biomass and their utilization for biofuel production.Plant biotechnology journal · 2016Review
- High-level expression of thermostable cellulolytic enzymes in tobacco transplastomic plants and their use in hydrolysis of an industrially pretreated Arundo donax L. biomass.Biotechnology for biofuels · 2016Article
- A Fusion between Domains of the Human Bone Morphogenetic Protein-2 and Maize 27 kD γ-Zein Accumulates to High Levels in the Endoplasmic Reticulum without Forming Protein Bodies in Transgenic Tobacco.Frontiers in plant science · 2016Article
- Transgenic tobacco plants with improved cyanobacterial Rubisco expression but no extra assembly factors grow at near wild-type rates if provided with elevated CO2.The Plant journal : for cell and molecular biology · 2016Article
- Rapid transient expression of human granulocyte-macrophage colony-stimulating factor in two industrial cultivars of tobacco (Biotechnology reports (Amsterdam, Netherlands) · 2015Article
- Cloning and Expression of TNF Related Apoptosis Inducing Ligand in Nicotiana tabacum.Iranian journal of pharmaceutical research : IJPR · 2015Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundChloroplast transformation in tobacco has been used extensively to produce recombinant proteins and enzymes. Chloroplast expression cassettes can be designed with different configurations of the cis-acting elements that govern foreign gene expression. With the aim to optimize production of recombinant hemicellulases in transplastomic tobacco, we developed a set of cassettes that incorporate elements known to facilitate protein expression in chloroplasts and examined expression and accumulation of a bacterial xylanase XynA. Biomass production is another important factor in achieving sustainable and high-volume production of cellulolytic enzymes. Therefore, we compared productivity of two tobacco cultivars - a low-alkaloid and a high-biomass - as transplastomic expression platforms.
resultsFour different cassettes expressing XynA produced various mutant phenotypes of the transplastomic plants, affected their growth rate and resulted in different accumulation levels of the XynA enzyme. The most productive cassette was identified and used further to express XynA and two additional fungal xylanases, Xyn10A and Xyn11B, in a high-biomass tobacco cultivar. The high biomass cultivar allowed for a 60% increase in XynA production per plant. Accumulation of the fungal enzymes reached more than 10-fold higher levels than the bacterial enzyme, constituting up to 6% of the total soluble protein in the leaf tissue. Use of a well-characterized translational enhancer with the selected expression cassette revealed inconsistent effects on accumulation of the recombinant xylanases. Additionally, differences in the enzymatic activity of crude plant extracts measured in leaves of different age suggest presence of a specific xylanase inhibitor in the green leaf tissue.
conclusionOur results demonstrate the pivotal importance of the expression cassette design and appropriate tobacco cultivar for high-level transplastomic production of recombinant proteins.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.