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

We focus on the development and use of synthetic tools to scrutinize the biology of the most abundant protein, the photosynthetic CO2-fixing enzyme Rubisco.

About

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.

Publications

Below is a list of selected publications. For a complete list of publications on the ISI website.

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.

Projects

Projects are availble aimed at discovering fundamental issues relating to the biogenesis and catalytic function of the CO2-fixing enzyme Rubisco.

Theme

Photosynthesis and plant energy biology

Student intake

Open for Bachelor, Honours, PhD students

Status

Current

People

Temperature determines species distributions and affects functions, reproduction, and survival. This project will assess the thermal sensitivity of different life stages and plant tissues to heat stress and will develop models to assess how Australian plant species will respond to a warmer and more extreme climate.

Theme

Behavioural, evolutionary and physiological ecology, Photosynthesis and plant energy biology, Plant environmental biology and functional ecology

Student intake

Open for Summer scholar, Honours, PhD students

Status

Current

People

Genetic, biochemical and physiological tools are being used to explore which residues in Rubisco and its chiropractic helper protein, Rubisco activase, that influence their interactions.

Theme

Photosynthesis and plant energy biology

Status

Current

People

During the last decade the practice of laboratory-directed protein evolution has become firmly established as a versatile tool in biochemical research by enabling molecular evolution towards desirable phenotypes or detection of novel structure-function interactions (see Bershtein & Tawfik, 2008, Curr Opin Chem Biol 12:

Theme

Photosynthesis and plant energy biology

Student intake

Open for PhD students

Status

Current

People

Examining how plant photosynthesis and growth is influenced by transplanting foriegn or modified Rubisco into higher plant chloroplasts.

Theme

Plant genetics and gene regulation, Photosynthesis and plant energy biology

Members

Group Leader

Senior Research Officer

Sarah Kaines

Senior Technical Services Officer#Plant Sciences

PhD Student

Honorary Lecturer

Divisional Visitor

Volunteer

News

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