summer research 2012 studying the metabolic pathway of acidobacterium capsulatum sunny ketchum,...
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Summer Research 2012
Studying the metabolic pathway of Acidobacterium Capsulatum
Sunny Ketchum, Pariss Trujillo, and Professor Les Sommerville
0 20 40 60 80 100 120 140 160 1800.00E+00
2.00E-01
4.00E-01
6.00E-01
8.00E-01
1.00E+00
1.20E+00
1.40E+00
1.60E+00
1.80E+00
OD 600
mg glucose/mL
Time (hours)
Gro
wth
Par
amet
ers
([gl
ucos
e], O
D 6
00)
Figure 1.- A. Capsulatum growth and glucose concentration
It was found that A.Cap grew on one sole carbon source, glucose, at an exponential rate until all glucose was consumed.
0 20 40 60 80 100 120 140 160 1800
0.5
1
1.5
2
2.5
3
3.5
Abs @ 600 nm
Protein/450
pH level
Time in hours
[Pro
tein
] (ug
/mL
* 45
0)Tu
rbid
ity
[abs
. @60
0nm
]pH
leve
l
Figure 2.- Turbidity, pH, and protein concentration as a function of time in hours
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.
* 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)
1500 mg glucose in media at zero hours
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.
H’NMR spectra of A.Cap in exponential phase
H’NMR spectra of A.Cap in stationary phase