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Abstract Detail


Metabolism

Bestman, Hank D. [1], Mobach, Janina [2], Zee, Jenna [2], Booy, Amanda [2].

The influence of the growing conditions on the polar metabolite fingerprint of Chlamydomonas reinhardtii as determined by 1H-NMR.

As part of a systems biology approach to studying metabolism 1H-NMR spectroscopy can be used as a rapid screen to characterize differences in metabolite fingerprints prior to obtaining more detailed metabolite profiles by 2-D NMR or GC-MS. We determined the effect of growing conditions on the polar metabolite fingerprint of the alga Chlamydomonas reinhardtii. The algae (wild type strain CC-125 mt+) are grown autotrophically under light (300 umol/m2/sec PAR) in continuous culture systems (160 mL) with a 2-L/min airflow (1500 ppm CO2). Heterotrophic conditions are created by adding sodium acetate to the nutrient medium (25 mM final concentration) and placing the culture system in the dark. This induces the glyoxylate shunt enabling the algae to synthesize carbohydrates from acetate. Following treatment the algal cells are collected, frozen in liquid nitrogen, and freeze dried. Algal metabolites are extracted in a NMR solvent (8:2 D2O:CD3OD; Phytochemistry, 2003, 62:949-957). The NMR spectra are processed unsupervised and reduced to integrated regions of equal width (0.01 ppm). Treatment effects are determined by Principal Component Analysis. When Chlamydomonas is grown heterotrophically for 6 hrs with acetate as its carbon source the metabolite fingerprints differ significantly from those of algae growing under autotrophic conditions. PCA-generated biplots show that changing the growing conditions from autotrophic to heterotrophic is accompanied by significant changes in intensity at the 1.3, 1.89-1.91, 3.60, 3.73, 3.89, 3.94 ppm chemical shift regions of the spectrum.


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1 - The King's University College, Biology and Center for Molecular Structure, 9125 50 Street, Edmonton, Alberta, T6B 2H3, Canada
2 - The King's University College, Biology

Keywords:
metabolite fingerprint
Chlamydomonas
glyoxylate shunt
NMR.

Presentation Type: Plant Biology Abstract
Session: P
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM
Number: P19013
Abstract ID:393


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