Team:UC Davis/Protein Engineering MM

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<h3 style="color:#212f20;margin-bottom: 0;">Assay Components</h3>
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<h3 style="color:#212f20;margin-bottom: 0;">Experimentally Derived Kinetic Constants for Our Aldehyde Dehydrogenases</h3>
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Now that we had identified three aldehyde dehydrogenases with unique specificity profiles, we proceeded to experimentally determine the kinetic constants (kcat and KM) of these enzymes on each substrate in our assay conditions.<br><br>
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<b>Michaelis-Menten</b>
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Now that we had identified four aldehyde dehydrogenases with unique specificity profiles, we proceeded to experimentally determine the kinetic constants (kcat and KM) of these enzymes on each substrate in our assay conditions.<br><br>
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Revision as of 18:13, 14 October 2014

UC Davis iGEM 2014

Design

Design

Build

Build

Test

Test

Experimentally Derived Kinetic Constants for Our Aldehyde Dehydrogenases

Now that we had identified three aldehyde dehydrogenases with unique specificity profiles, we proceeded to experimentally determine the kinetic constants (kcat and KM) of these enzymes on each substrate in our assay conditions.

Kinetic Constants of WT Escherichia coli Aldehyde Dehydrogenase
Substratekcatkcat ErrorKMKM Errorkcat/KMkcat/KM Error
Propionaldehyde8.000.27782.8750.8310221.04748.00
Butyraldehyde6.270.28196.6827.6531867.744700.95
Pentanal6.180.2245.926.77134598.0020398.49
Hexanal5.370.5312.656.36424348.82217516.13
HeptanalBad Fit---------------
Octanal4.530.6140.3323.41112265.5266927.82
Nonanal3.990.4427.6214.10144584.4275534.70
DecanalBad Fit---------------
E-2-Pentenal0.940.0599.3320.749480.512054.66
E-2-Hexenal1.290.0431.164.8441478.416599.45
E-2-Octenal0.990.0312.271.8681100.4712529.78
E-2-Nonenal0.870.029.601.1690621.9311127.45
E-2-Decenal0.660.0113.341.3749715.955217.05
E,E-2,4-Heptadienal0.680.11142.0875.994809.312682.89
E,E-2,4-NonadienalBad Fit---------------
E,E-2,4-DecadienalNDNDNDNDNDND
Kinetic Constants of W176V Escherichia coli Aldehyde Dehydrogenase
Substratekcatkcat ErrorKMKM Errorkcat/KMkcat/KM Error
Propionaldehyde7.571.242575.71559.822937.45800.31
Butyraldehyde4.420.12532.7032.008306.37546.34
Pentanal5.440.35165.3535.2532877.937328.72
Hexanal7.290.3738.938.48187159.1441828.31
Heptanal5.870.3826.177.87224363.8568987.72
Octanal6.550.4584.2521.4177758.6620464.40
Nonanal6.410.3175.2913.5485145.0415837.70
Decanal4.790.1847.937.3299954.3115711.24
E-2-PentenalNDNDNDNDNDND
E-2-Hexenal0.220.0228.119.857797.242796.55
E-2-OctenalBad Fit---------------
E-2-NonenalBad Fit---------------
E-2-DecenalBad Fit---------------
E,E-2,4-HeptadienalNDNDNDNDNDND
E,E-2,4-NonadienalNDNDNDNDNDND
E,E-2,4-DecadienalNDNDNDNDNDND
Kinetic Constants of WT Rattus norvegicus (Rat) Aldehyde Dehydrogenase
Substratekcatkcat ErrorKMKM Errorkcat/KMkcat/KM Error
Propionaldehyde19.900.511002.3645.0219856.171026.79
Butyraldehyde12.460.22261.0213.6047740.022629.46
Pentanal10.510.44118.0517.2889060.7913565.38
Hexanal8.280.6632.9011.62251744.0991148.58
HeptanalBad Fit---------------
Octanal6.620.57107.9432.6761315.1819282.67
Nonanal7.620.6888.6929.0285930.0629145.22
Decanal9.060.6693.8424.9296541.8026598.58
E-2-Pentenal8.680.54694.0586.7512509.321745.41
E-2-Hexenal8.540.15220.1411.5438785.272139.18
E-2-Octenal7.510.4765.8116.08114129.5628807.70
E-2-Nonenal9.500.5875.4117.52125987.9430284.84
E-2-Decenal6.050.2558.699.63103066.4517434.71
E,E-2,4-Heptadienal6.010.3131.777.40189320.3945208.07
E,E-2,4-Nonadienal6.440.4749.3814.83130374.0340304.11
E,E-2,4-Decadienal6.320.3330.757.27205360.2549735.07

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