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This study delves into the metabolic pathway of Acidobacterium Capsulatum, specifically its growth on glucose as a sole carbon source. The research uncovers the exponential growth pattern of A. Cap until glucose is fully consumed, with peak growth observed at 75 hours. Turbidity, pH, and protein concentration over time are analyzed, revealing valuable insights. Additionally, energy conservation in anaerobic bacteria and biomass estimates are discussed, including polysaccharide excretion and glucose utilization for energy. Quenching and extraction techniques using NaCl and perchloric acid are detailed for extracting water-soluble metabolites and enhancing H’NMR spectra analysis at different growth phases.
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Summer Research 2012 Studying the metabolic pathway of AcidobacteriumCapsulatum Sunny Ketchum, Pariss Trujillo, and Professor Les Sommerville
It was found that A.Cap grew on one sole carbon source, glucose, at an exponential rate until all glucose was consumed. Figure 1.- A. Capsulatumgrowth and glucose concentration
A.Cap reached its peak growth after 75 hours. This displays the amount of protein found in A.Cap along with the growth environment after a certain amount of hours. Figure 2.- Turbidity, pH, and protein concentration as a function of time in hours
1500 mg glucose in media at zero hours * Thauer RK, Jungerman K, Decker K. Energy Conservation in Chemotrophic Anaerobic Bacteria. Bacteriological Reviews (41)1, 1977, 100-180. Biomass Estimate From Protein Concentration 600 mg dry weight (assuming 15% protein and 1% inorganic ions by wet cell weight) 1500 mg – 600 mg – 600 mg = 300 mg POLYSACCHARIDE EXCRETED .9 g x (1 mol ATP/9 g cells) x (1 mole glucose/30 moles ATP) x (180 g glucose/mole) x (1000 mg/1 g) = 600 mg GLUCOSE USED FOR ENERGY * (including energy needed for polysaccharide synthesis)
Quenching and Extraction of Water Soluble Metabolites A series of NaCl and 35% per chloric acid rinses were used on the cells in order to quench and extract the water soluble metabolites. This gave a clearer H’NMR spectra. The H’NMR peaks were then identified for metabolites.
