Summer Research 2012 Studying the metabolic pathway of Acidobacterium Capsulatum Sunny Ketchum,...

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