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E. Coli       Cyano       Enzymology       Media

Week 1 - June 1-4
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E. Coli
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Cyano
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Hexokinase coupled assay used to quantitate amounts of cyanobacteria glucose secretion.

Rxn:
• 1. glucose + ATP --Hx-> G-6-P + ADP
• 2. G-6-P + NAD+ --G-6-PDeH-> G-6-P + NADH

Reaction used Glucose assay kit (Genzyme) containing 2000U/L Hexokinase reagent and 4000U/L G-6-P DeH and measured glucose concentrations ranging from 0.05mM-0.25mM and absorbances were measured at 340.0 nm.

Results: Final NADH concentrations were determined using the molar extinction coefficient and were proportionate to initial glucose concentrations.
Discussion: Focus is now to measure not only glucose, but fructose concentrations as well as both with be secreted in the form of sucrose using a D-glucose/D-fructose assay (Megaenzyme.

Media

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