From Pandemic Preparedness to Biofuel Production: Tobacco Finds Its Biotechnology Niche in North America

Powell, Joshua D.

doi:10.3390/agriculture5040901

Cited by 18 publications

(13 citation statements)

References 120 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In this project we used the tobacco transient expression system, which allows for fast heterologous protein expression and is widely used as a way to screen combinatorial testing 32,31 . Because of the fast experimental turnover of the method, no obvious interference with the endogenous Tat system was observed.…”

Section: Discussion and Perspectivesmentioning

confidence: 99%

“…We did not attempt stable tobacco transformation since previous studies show that this is not a suitable system to express the dhurrin pathway 41 . Further, while not very prevalent yet, the transient expression system is becoming a common protein production platform system for commercial uses, especially for antibodies production and for vaccines such as ebola and influenza 31,42,43 .…”

Section: Discussion and Perspectivesmentioning

confidence: 99%

“…Transient infiltration of plants allows for high protein expression and is a quick and simple method for combinatorial studies 31,32 . Agrobacterium cells transformed with the expression vectors carrying single constructs were generated.…”

Section: Design Of Fusion Proteinsmentioning

confidence: 99%

See 2 more Smart Citations

Tat proteins as novel thylakoid membrane anchors organize a biosynthetic pathway in chloroplasts and increase product yield 5-fold

Jesus

Nielsen

Mellor

et al. 2017

Metabolic Engineering

View full text Add to dashboard Cite

Photosynthesis drives the production of ATP and NADPH, and acts as a source of carbon for primary metabolism. NADPH is also used in the production of many natural bioactive compounds. These are usually synthesized in low quantities and are often difficult to produce by chemical synthesis due to their complex structures. Some of the crucial enzymes catalyzing their biosynthesis are the cytochromes P450 (P450s) situated in the endoplasmic reticulum (ER), powered by electron transfers from NADPH. Dhurrin is a cyanogenic glucoside and its biosynthesis involves a dynamic metabolon formed by two P450s, a UDP-glucosyltransferase (UGT) and a P450 oxidoreductase (POR). Its biosynthetic pathway has been relocated to the chloroplast where ferredoxin, reduced through the photosynthetic electron transport chain, serves as an efficient electron donor to the P450s, bypassing the involvement of POR. Nevertheless, translocation of the pathway from the ER to the chloroplast creates other difficulties, such as the loss of metabolon formation and intermediate diversion into other metabolic pathways. We show here that co-localization of these enzymes in the thylakoid membrane leads to a significant increase in product formation, with a concomitant decrease in off-pathway intermediates. This was achieved by exchanging the membrane anchors of the dhurrin pathway enzymes to components of the Twin-arginine translocation pathway, TatB and TatC, which have self-assembly properties. Consequently, we show 5-fold increased titers of dhurrin and a decrease in the amounts of intermediates and side products in Nicotiana benthamiana. Further, results suggest that targeting the UGT to the membrane is a key factor to achieve efficient substrate channeling.

show abstract

Section: Discussion and Perspectivesmentioning

confidence: 99%

Section: Discussion and Perspectivesmentioning

confidence: 99%

See 1 more Smart Citation

Tat proteins as novel thylakoid membrane anchors organize a biosynthetic pathway in chloroplasts and increase product yield 5-fold

Jesus

Nielsen

Mellor

et al. 2017

Metabolic Engineering

View full text Add to dashboard Cite

show abstract

“…The tobacco species Nicotiana benthamiana is often used to express monoclonal antibodies and vaccine antigens. This expression in possible through the delivery of engineered genetic constructs from Agrobacterium tumefaciens bacteria (Powell 2015;Rybicki 2014). The delivery of A. tumefaciens cultured in bacterial broth medium into tobacco is referred to as agroinfiltration.…”

Section: Introductionmentioning

confidence: 99%

A cytometry microparticle platform approach for screening tobacco microRNA changes after agrobacterium delivery

Powell

Chen

Mason

2016

Journal of Microbiological Methods

View full text Add to dashboard Cite

MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens "agroinfiltration" expression strategy. A multiplex microparticle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach.

show abstract

“…Consequently, it would be difficult to meet the antibody demand for widespread use in Ebola virus-stricken African countries. Moreover, the drug was originally intended for expression at levels sufficient for animal trial use 36 . In addition, ZMapp is yet to receive approval by the US Food and Drug Administration who would have to certify, for instance, that the plant extraction process has not led to contamination of the resulting drug 37 .…”

Section: Contemporary Application: Treatment Of Ebola Patientsmentioning

confidence: 99%

Plantibodies in human and animal health: a review

Oluwayelu

Adebiyi

2016

Afr H. Sci.

View full text Add to dashboard Cite

Background: Antibodies are essential part of vertebrates' adaptive immune system; they can now be produced by transforming plants with antibody-coding genes from mammals/humans. Although plants do not naturally make antibodies, the plant-derived antibodies (plantibodies) have been shown to function in the same way as mammalian antibodies. Methods: PubMed and Google search engines were used to download relevant publications on plantibodies in medical and veterinary fields; the papers were reviewed and findings qualitatively described. Results: The process of bioproduction of plantibodies offers several advantages over the conventional method of antibody production in mammalian cells with the cost of antibody production in plants being substantially lesser. Contrary to what is possible with animal-derived antibodies, the process of making plantibodies almost exclusively precludes transfer of pathogens to the end product. Additionally, plants not only produce a relatively high yield of antibodies in a comparatively faster time, they also serve as cost-effective bioreactors to produce antibodies of diverse specificities. Conclusion: Plantibodies are safe, cost-effective and offer more advantages over animal-derived antibodies. Methods of producing them are described with a view to inspiring African scientists on the need to embrace and harness this rapidly evolving biotechnology in solving human and animal health challenges on the continent where the climate supports growth of diverse plants.

show abstract

From Pandemic Preparedness to Biofuel Production: Tobacco Finds Its Biotechnology Niche in North America

Cited by 18 publications

References 120 publications

Tat proteins as novel thylakoid membrane anchors organize a biosynthetic pathway in chloroplasts and increase product yield 5-fold

Tat proteins as novel thylakoid membrane anchors organize a biosynthetic pathway in chloroplasts and increase product yield 5-fold

A cytometry microparticle platform approach for screening tobacco microRNA changes after agrobacterium delivery

Plantibodies in human and animal health: a review

Contact Info

Product

Resources

About