Whitney Group - Synthetic Photosynthesis - bioengineering enzymes to adjust carbon fixation

Our research focuses on the development and use of synthetic biology tools to scrutinize the biology of the biospheres most abundant protein, the photosynthetic CO2-fixing enzyme Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase). The research provides training in molecular biology, protein engineering (directed evolution), enzyme kinetics, biochemistry, plastome transformation, tissue culture, measuring leaf photosynthesis and plant growth.

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Selected publications

Some recent research papers

Sharwood, R, Crous, K, Whitney, S.M et al (2017) 'Linking photosynthesis and leaf N allocation under future elevated CO2 and climate warming in Eucalyptus globulus', Journal of Experimental Botany, vol. 68, 1157-1167.

Heureux, A, Young, J, Whitney, S.M et al (2017) 'The role of Rubisco kinetics and pyrenoid morphology in shaping the CCM of haptophyte microalgae', Journal of Experimental Botany, vol. Online, doi: 10.1093/jxb/erx179.

Sharwood, R, Sonawane, B, Ghannoum, O, Whitney, S.M (2016) 'Improved analysis of C4 and C3 photosynthesis via refined in vitro assays of their carbon fixation biochemistry', Journal of Experimental Botany, vol. 67, no. 10, pp. 3137-3148.

Young, J, Heureux, A, Sharwood, R, Whitney, S.M (2016) 'Large variation in the Rubisco kinetics of diatoms reveals diversity among their carbon-concentrating mechanisms',  Journal of Experimental Botany, vol. 67, no. 11, pp. 3445-3456.

Sharwood, R, Ghannoum, O & Whitney, S.M (2016) 'Prospects for improving CO2 fixation in C3-crops through understanding C4-Rubisco biogenesis and catalytic diversity', Current Opinion in Plant Biology, vol. 31, pp. 135-142.

Sharwood, R, Ghannoum, O, Kapralov, M, Gunn, L, Whitney, S.M (2016) 'Temperature responses of Rubisco from Paniceae grasses provide opportunities for improving C3 photosynthesis', Nature Plants, vol. 2, no. 16186, pp. 1-9.

Wilson, R, Alonso, H & Whitney, S.M (2016) 'Evolving Methanococcoides burtonii archaeal Rubisco for improved photosynthesis and plant growth', Scientific Reports, vol. 6, Article 22284.

Whitney, S.M, Birch, R, Kelso, C et al (2015) 'Improving recombinant Rubisco biogenesis, plant photosynthesis and growth by coexpressing its ancillary RAF1 chaperone', Proceedings of the National Academy of Sciences of the United States of America, vol. 112, no. 11, pp. 3564-3569.

Some reviews

Bracher, A, Whitney, S.M, Hartl, F et al (2017) 'Biogenesis and metabolic maintenance of Rubisco', Annual Review of Plant Biology, vol. 68, pp. 29-60.

Wilson, R & Whitney, S.M (2015) 'Photosynthesis: Getting it together for CO2 fixation', Nature Plants, vol. 1, pp. 15130.

Parry, M, Andralojc, P, Scales, J, Salvucci, ME, Carmo-Silva, E, Alonso, H, Whitney, SM. (2013) 'Rubisco activity and regulation as targets for crop improvement', Journal of Experimental Botany, vol. 64, 717-730

Whitney S.M., Houtz R.L and Alonso H (2011) Advancing our understanding and capacity to engineer nature’s CO2 sequestering enzyme, Rubisco.  Plant Physiology 155, 27-35

Andrews, T.J. and Whitney, S.M. (2003) Manipulating Rubisco in the chloroplasts of higher plants. Archives of Biochemistry and Biophysics 414: 159-169

Synthetic biology in E. coli - Directed evolution of Rubisco

Wilson, R & Whitney, S.M (2017) 'Improving CO2 Fixation by Enhancing Rubisco Performance', in Miguel Alcalde (ed.), Directed Enzyme Evolution: Advances and Applications, Springer, Spain, pp. 101-126

Wilson, R, Alonso, H & Whitney, S.M (2016) 'Evolving Methanococcoides burtonii archaeal Rubisco for improved photosynthesis and plant growth', Scientific Reports, vol. 6, Article 22284

Mueller-Cajar and Whitney S.M. (2008) Directing the evolution of Rubisco and Rubisco activase - first impressions of a new tool for photosynthesis research. Photosynthesis Research, 98, 667-675

Mueller-Cajar and Whitney S.M. (2008) Evolving improved Synechococcus Rubisco functional production in Escherichia coli. Biochemical Journal,414, 201-214

Mueller-Cajar O, Morell M, Whitney S.M. (2007) Directed evolution of Rubisco in E. coli reveals a specificity-determining hydrogen bond in the Form II enzyme. Biochemistry, 46, 14067-74

Greene D.N, Whitney S.M, Matsumura I. (2007) Artificially evolved Synechococcus PCC6301 Rubisco variants exhibit improvements in folding and catalytic efficiency. Biochemical Journal, 404, 517-24

Synthetic biology in chloroplasts - engineering of Rubisco and Rubisco activase

von Caemmerer, S, Tazoe, Y, Evans, J, Whitney S.M (2014) 'Exploiting transplastomically modified Rubisco to rapidly measure natural diversity in its carbon isotope discrimination using tuneable diode laser spectroscopy', Journal of Experimental Botany, 65, pp. 3759-3767.

Whitney, S & Sharwood, R (2014), 'Plastid transformation for Rubisco engineering and protocols for assessing expression.', in Pal Maliga (ed.), Chloroplast Biotechnology. Methods and Protocols, Humana Press, New York, pp. 245-262.

Sharwood, R & Whitney, S (2014) 'Correlating Rubisco catalytic and sequence diversity within C3 plants with changes in atmospheric CO2 concentrations', Plant Cell and Environment, 37, 1981-1984.

Pengelly, J, Forster, B, von Caemmerer, S, Badger M, Price GD & Whitney SM (2014) 'Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts', Journal of Experimental Botany, 65, 3071-3080.

Galmes, J, Perdomo, J, Flexas, J and Whitney, SM. (2013) 'Photosynthetic characterization of Rubisco transplantomic lines reveals alterations on photochemistry and mesophyll conductance', Photosynthesis Research, 115, 153-166.

Price, G, Pengelly, J, Forster, B, von Caemmerer, S, Badger, MR, Price, GD and Whitney, SM. (2013) 'The cyanobacterial CCM as a source of genes for improving photosynthetic CO2 fixation in crop species', Journal of Experimental Botany, 64, 753-768.

Whitney, S.M Sharwood, R.E, Orr, D White, S.J, Alonso, H and Galmes, J (2011) Isoleucine 309 acts as a C4 catalytic switch that increases ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylation rate in Flaveria. Proceedings of the National Academy of Sciences 108, 14688-14693.

Blayney, M, Whitney, S.M and Beck, J (2011) NanoESI mass spectrometry of Rubisco and Rubisco activase structures and their interactions with nucleotides and sugar phosphates'. Journal of the American Society for Mass Spectrometry 22, 1588-1601.

Alonso H, Blayney M.J, Beck J.L and Whitney S.M (2009) Substrate induced assembly of Methanococcoides burtonii D-Ribulose-1,5-Bisphosphate carboxylase/oxygenase dimers into decamers. Journal of Biological Chemistry 284, 33876-82

Whitney S.M and Sharwood R.E (2008) Construction of a tobacco master line to improve Rubisco engineering in chloroplasts. Journal of Experimental Botany 59, 1909-1921

Sharwood R.E, von Caemmerer S, Maliga P and Whitney S.M. (2008) The catalytic properties of hybrid Rubisco comprising tobacco small and sunflower large subunits mirror the kinetically equivalent source Rubiscos and can support tobacco growth. Plant Physiology 146, 83-96

Whitney, S.M and Sharwood R.E (2007). Linked Rubisco subunits can assemble into functional oligomers without impeding catalytic performance. Journal of Biological Chemistry 282, 3809-3818

Rubisco bioengineering in chloroplasts using synthetic biology– where it all began

Whitney, S.M. and Andrews, T.J. (2003) Photosynthesis and growth of tobacco with a substituted bacterial Rubisco mirror the properties of the introduced enzyme Plant Physiology 133, 287-294

Whitney, S.M. and Andrews, T.J (2001) Plastome-encoded bacterial ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) supports photosynthesis and growth in tobacco. Proceeding of the National Academy of Sciences 98: 14738-14743.

Whitney, S. M. and T. J. Andrews (2001) The gene for the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit relocated to the plastid genome of tobacco directs the synthesis of small subunits that assemble into Rubisco. Plant Cell 13: 193-205.

Whitney, S.M., von Caemmerer, S., Hudson, G.S. and Andrews, T.J. (1999) Directed mutation of the Rubisco large subunit of tobacco influences photorespiration and growth. Plant Physiology, 121, 579-588.

All publications

PS PhD Exit Seminar: Pathways to engineer plant Rubisco

Event | Fri 19 August 2022
In plants, Rubisco is responsible for the assimilation of CO2 during photosynthesis.

PS Seminar Series- Enhancing Rubisco catalysis improves plant growth

Event | Wed 30 March 2022
The rate of photosynthesis and plant growth is often rate limited by the activity of the CO2-fixing enzyme Ribulose bisphosphate carboxylase oxygenase (Rubisco).

BSB PhD Exit Seminar: Structure-function studies of multidrug resistance transporters of the malaria parasite

Event | Thu 18 November 2021
Many genes that have been recently identified as determinants of drug resistance in the malaria parasite encode membrane transport proteins (also known as transporters). 

PS Webinar Series- PhD Exit Seminar - Evolving red-type Rubisco towards enhancing plant carbon fixation

Event | Fri 29 January 2021
I have developed a novel E.coli based directed evolution system to evolve Ribulose-1,5 -bisphosphate carboxylase/oxygenase (Rubisco), the rate-limiting enzyme in the Calvin Cycle of photosynthetic organisms.
Ding Yuan

PS PhD Exit Seminar: Monocot Rubisco meets Dicot Chloroplast: A Structure-Function Relationship Study

Event | Fri 27 September 2019
Explore the elimination of the folding/assembly incompatibility between the monocot crop (rice, wheat and maize) Rubisco and dicot model plant (tobacco) chloroplasts through a mutagenesis strategy which introduces certain amino acid substitutions into the large subunit of the monocot Rubisco.

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