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Nicholas C Carpita

Botany and Plant Pathology 

  • Professor
765.494.4653
765.494.0363
Lilly Hall Room 1-464

Cellulose structure and biosynthesis   With more than 1011 tons made annually in the natural environment, cellulose is the most abundant biopolymer on Earth. Cellulose microfibrils, the fundamental scaffolding of the plant cell wall, is a para-crystalline array of several dozen (1®4)-b-D-glucan chains synthesized at the plasma membrane surface by large multicomponent complexes of cellulose synthase (CesA) proteins. We discovered that recombinant catalytic domains of CesA are two-domain structures that dimerize using Small-Angle X-ray Scattering (SAXS) experiments to derive 3-D surface contour structures (Olek et al. 2014). The catalytic domains of plant CesAs contain two unique sequences not found in prokaryotic ancestors ­– a Plant-Conserved Region (P-CR) and Class-Specific Region (CSR) of unknown function. Molecular docking experiments with the catalytic core predicted that the CSRs of CesAs are the dimerization domains. We aim to provide the first crystal structure of a plant CesA catalytic domain. Towards that goal, we crystallized a recombinant Plant-Conserved Region (P-CR) and showed that it is primarily a coiled-coil domain positioned near the entrance to the active site of the catalytic core (Rushton et al. 2016). With Wen Jiang (Purdue) we have begun studies to define the assembly of CesAs into complexes at the Golgi membrane as part of a broader effort to characterize the dynamics of the Golgi proteome.  From substrate binding stoichiometry, we know that each CesA protein synthesizes a single (1®4)-b-D-glucan chain of a microfibril (Olek et al. 2014). We developed a modified TEMPO-catalyzed oxidation of glucose to uronosyl residues followed by carboxyl reduced with NaBD4 to provide a novel method for determining proportion of disordered surfaced chains, ordered surface chains in which only one-half of the residues are exposed to water, and completely anhydrous core-crystalline residues for cellulose microfibrils in biomass. Microfibrils of primary or secondary walls from seedlings and lignocellulosic biomass have significantly higher content of their glucan chains in anhydrous domains, indicating cellulose microfibrils into bundles with extensive crystalline continuity.

 

Cell-wall gene discovery  - For several decades, my lab focused on determining the fine structure and dynamics of synthesis and degradation of the non-cellulosic polysaccharides of the plant cell wall. This work established that flowering plants synthesize two distinct kinds of primary cell walls – a ‘Type I’ wall made by most dicots and about one-half of monocots, and a ‘Type II’ wall made by grasses and closely related ‘commelinid’ monocots (Carpita and Gibeaut, 1993). This work has had a major impact on fashioning perspectives for how the unique chemical structure of grasses impacts technologies for their conversion to biofuels and bio-based products. Our discovery of unique arabinoxylan and rhamnogalacturonan I structures in seed mucilages not seen in growing cell walls supports a hypothesis that plants make a selected few ubiquitous backbone polymers onto which a broad spectrum of side group substitutions are added to engender many possible functions. We have annotated almost 1500 genes that function in wall biogenesis in Arabidopsis, rice, and maize, a model C4 grass, assembling them into 74 gene families (Penning et al. 2009). We continue to expand annotations of the many gene families of cell-wall related genes for grasses and other angiosperms (see http://cellwall.genomics.purdue.edu). Work with colleagues characterized the functions of numerous previously unknown cell-wall genes through analysis of fine structure of alterations in specific polysaccharides in mutants. With colleagues from the National Renewable Energy Lab, we used Pyrolysis Molecular Beam MS to define fragments diagnostic of lignin monomers, hydroxycinnamic acids, xylans and cellulose composition in parallel to high throughput saccharification assays to define variation in glucose and xylose yield

Awards & Honors

(2013) Fellow of the American Association for the Advancement of Science. American Association for the Advancement of Science.

(2012) Fellow of AAAS. American Association for the Advancement of Science.

(2012) Immediate Past President. American Society of Plant Biologists.

(2010) President of American Society of Plant Biologist. American Society of Plant Biologist.

(2009) Seeds of Success Award, Millionaire's Club. College of Agriculture.

(1991) Agricultural Research Award. Purdue University.

(1987) Gamma Sigma Delta. Purdue University.

(1984) Sigma Xi. Purdue University.

Patents

Carpita, N. C. none.

Selected Publications

McCann, M., & Carpita, N. (in press). Biomass recalcitrance: A multi-scale, multi-factor and conversion-specific property. Journal of Experimental Botany.

Carpita, N., & McCann, M. (in press). Characterizing visible and invisible cell-wall mutant phenotypes. Journal of Experimental Botany.

Penning, B., Sykes, R., Babcock, N., Dugard, C., Held, M., Klimek, J., . . . Carpita, N. (2014). Genetic determinants for enzymatic digestion of lignocellulosic biomass are independent of those for lignin abundance in a maize recombinant inbred population. Plant Physiology, 165, 1475-1487.

Carpita, N. (2014). No carbon left behind: A new paradigm in the conversion of biomass to biofuels and high-value products. International Bioenery Symposium. Retrieved from http://bioenergy2014.co.uk/

Carpita, N. (2014). Redesigning the plant cell wall for conversion of biomass to biofuels and high-value products. Plenary Lecture, Frontiers in Biorefining. Retrieved from http://www.frontiersinbiorefining.org/

Baldwin, L., Domon, J., Klimek, J., Sellier, H., Gillet, G., Pelloux, J., . . . Rayon, C. (2014). Structural alteration of cell wall pectins accompanies pea development in response to cold. Phytochemistry, 104, 37-47.

McCann, M., Penning, B., Dugard, C., & Carpita, N. (in press). Tailoring plant cell wall composition and architecture for conversion to liquid hydrocarbon biofuels. In Direct Microbial Conversion of Biomass to Advanced Biofuels.

Vinueza, N., Kim, E., Gallardo, V., Mosier, N., Abu-Omar, M., Carpita, N., & Kenttamaa, H. (in press). Tandem mass spectrometric characterization of conversion of xylose to furfural. Biomass Bioenergy.

Olek, A., Rayon, C., Makowski, L., Ciesielski, P., Badger, J., Paul, L., . . . Carpita, N. (2014). The structure of the catalytic domain of a plant cellulose synthase and its assembly into dimers. Plant Cell, 26, 2996-3009.

Rayon, C., Olek, A., & Carpita, N. (2014). Towards redesigning cellulose biosynthesis for improved bioenergy feedstocks. In Plants and BioEnergy, Advances Plant Biology (4, 183-193). New York: Springer.