ShyamMasakapalli.gif
Mr Shyam Masakapalli M.Sc, M.Tech (IIT-Kgp)
Currently Postdoc assistant with Dr Lee Sweetlove, D.Phil was with Dr Nick Kruger and Prof. George Ratcliffe

Exeter College

E-mail:
  shyam.masakapalli@plants.ox.ac.uk

Website:
  www.epernicus.com/skm6

Tel  +44 (0)1865 275143

Last Modified: April 2012

Mr SK Masakapalli

Research Area

Network flux analysis of central and secondary metabolism in plants

Research Description

Network flux analysis aims to generate flux maps of the metabolic network and thus to provide a functional description of the metabolic phenotype of the organism under study. Steady state flux analysis, based on stable isotope labelling experiments, is one of the principal experimental methods in network flux analysis and its application to the plant metabolic network is increasing.


Postdoctoral Research: (Supervisors: Dr Lee Sweetlove and Prof. George Ratcliffe)

Project: “EU-Smart Cell Project: Rational design of plant systems for sustainable generation of value added industrial products” see weblink http://www.smart-cell.org/

Specific aims: To establish flux maps of primary and secondary metabolism in transgenic hairy root lines of Tobacco and Catharanthus roseus. The focus is towards the sustainable generation of value added industrial products.

D.Phil Research aims and achievements: (Supervisors: Prof. George Ratcliffe and Dr Nick Kruger)

Thesis title: Network flux analysis of central metabolism in plants

The aim of the project was to establish and validate robust techniques for quantifying fluxes through the dominant pathways of carbohydrate oxidation in plants. The subcellular compartmentation of these pathways between the cytosol and plastid is a characteristic feature of the structure of the plant metabolic network, and the project focused on the extent to which network flux analysis is capable of distinguishing fluxes between and through the duplicated steps and pathways. This lead to assessment of the response of the compartmented network to nutritional perturbations of phosphate stress and nitrogen sources.

Established in silico modeling strategies for 13C-based steady state flux analysis using 13CFLUX® and quantified the fluxes in the compartmented model of heterotrophic Arabidopsis central carbon metabolism (published in Plant Physiology. 152: 602-619, 2010). This study has physiological implications in establishing the fluxes through the parallel reactions in the cytosol and plastid and explains the provision of reducing power (NADPH) via oxidative steps of Pentose phosphate pathway.

 The robust strategies developed were applied to establish the response of Arabidopsis central network to nutritional perturbations with relevance to phosphate stress and nitrogen source(s). The results have established the metabolic phenotypes with distinguishable response. The work was presented as talks and posters in various international conferences (SEB Glasgow 2011-Pathways and fluxes session July 2-4, International conference on Systems Biochemistry, University of York, March 21-23, 2010 and Plant Systems Biology UK workshop, Nottingham, June 1-3, 2010). The manuscripts for publication of the Phosphate and Nitrogen work are under preparation.

Our research group at Oxford also undertook a study related to the stability of the flux distribution of Arabidopsis suspension under different oxygenation conditions (published in Plant Physiology. 148: 704-718, 2008). More recently, we published a research review on the strategies for investigating the plant metabolic network with steady-state metabolic flux analysis (Journal of Experimental Botany, Special issue on Pathways & Fluxes: Analysis of the Plant Metabolic Network 2012). In other collaborative projects with peers at John Innes Center, UK and University of Oxford, we work on metabolic modeling of Rhizobium leguminosarum and Pseudomonas syringae. The metabolic phenotypes obtained were presented as posters in few international conferences in 2011.

Publications

N.J. Kruger, S.K. Masakapalli and R.G. Ratcliffe (2012) Strategies for investigating the plant metabolic network with steady-state metabolic flux analysis: lessons from an Arabidopsis cell culture and other systems. Journal of Experimental Botany (online).

Masakapalli, S.K, le, Lay P, Huddleston, J.E, Pollock, N.L, Kruger, N.J, Ratcliffe, R George. (2010) Subcellular flux analysis of central metabolism in a heterotrophic Arabidopsis cell suspension using steady-state stable isotope labeling Plant Physiology. 152 (2): pp 602-619.

Williams, T.C.R, Miguet, L, Masakapalli, S.K, Kruger, N.J, Sweetlove, L.J, Ratcliffe, R.G. (2008) Metabolic network fluxes in heterotrophic arabidopsis cells: Stability of the flux distribution under different oxygenation conditions Plant Physiology. 148 (2): pp 704-718.

Based on my M.Tech project work at IIT Kharagpur (2007-08)

M. Shyam Kumar, P. K. Datta, S. Dutta Gupta (2009) In vitro evaluation of UV opacity potential of Aloe vera L. gel from different germplasms. J Nat Med 63(2):195–199

Full Publication List (while at this department)

Kruger, N.J, Masakapalli, S.K, Ratcliffe, R.G. (2012) Strategies for investigating the plant metabolic network with steady-state metabolic flux analysis: Lessons from an Arabidopsis cell culture and other systems Journal of Experimental Botany. 63 (6): pp 2309-2323.
doi:10.1093/jxb/err382.

Masakapalli, S.K, le, Lay P, Huddleston, J.E, Pollock, N.L, Kruger, N.J, Ratcliffe, R George. (2010) Subcellular flux analysis of central metabolism in a heterotrophic Arabidopsis cell suspension using steady-state stable isotope labeling Plant Physiology. 152 (2): pp 602-619.
doi:10.1104/pp.109.151316.

Williams, T.C.R, Miguet, L, Masakapalli, S.K, Kruger, N.J, Sweetlove, L.J, Ratcliffe, R.G. (2008) Metabolic network fluxes in heterotrophic arabidopsis cells: Stability of the flux distribution under different oxygenation conditions Plant Physiology. 148 (2): pp 704-718.
doi:10.1104/pp.108.125195.

Collaborators

Funding Source: Clarendon Scholarship, ORS award and Mr Krishna Pathak scholarship (Exeter college)

Skills

In silico modelling of metabolism (13CFLUX and other tool); In Vivo NMR (31P, 13C, 15N); 1H/13C- NMR, GC-MS; Computational skills- C, C++, Linux, Matlab ; Refworks- Bibliographic tool.