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Metadate
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NADH
lactate
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Manganese
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| Item | Value | ||||||||
| Experimentator | Tomas Fiedler | ||||||||
| Description | Measurements on kinetics of the LDH of S. pyogenes, influence of allosteric effectors and substrate concentration on rates | ||||||||
| Aim | Assessment of kinetic parameters of LDH to include in a catabolic model | ||||||||
| Strain information | M49 | ||||||||
| Strain | S. pyogenes | ||||||||
| Vessel type | cuvette | ||||||||
| Temperature (degree celcius) | 37 | ||||||||
| Volume (ml) | 1 | ||||||||
| pH | 6.8 | ||||||||
| Explanations | Data | ||||||||
| TOPIC | |||||||||
| Titel | Measurements on kinetics of the recombinant, purified LDH of S. pyogenes, influence of allosteric effectors and substrate concentration on rates | ||||||||
| Comments | some comments | ||||||||
| ASSAY | |||||||||
| Title | LDH kinetics | ||||||||
| Assay_type | enzyme kinetics | ||||||||
| Comments | |||||||||
| EXPERIMENT | |||||||||
| Experiment_description | |||||||||
| Experiment_type | Concentration series | ||||||||
| Experimentalist | Tomas Fiedler | ||||||||
| Begin_date | 2009-02-16T0:0:0+0000 | ||||||||
| User | Tomas Fiedler | ||||||||
| Group | SYSMOLAB | ||||||||
| ORGANISM | |||||||||
| Organism | S.pyogenes | ||||||||
| NCBI_ID | 294934 | ||||||||
| Strain | M49 | ||||||||
| Genotype | wild-type | ||||||||
| Phenotype | wild-type | ||||||||
| Experimental_conditions | |||||||||
| Item | pH | temperature | buffer | ||||||
| Start_value | 6.8 | 37 | 50 mM NaPO4 | ||||||
| End_value | 6.8 | 37 | |||||||
| Unit | dimensionlesss | ºC | |||||||
| Comments | |||||||||
| FACTORS_STUDIED | |||||||||
| Item | Effect of substrate concentration on enzyme rate | ||||||||
| Start_value | 0 | ||||||||
| End_value | 100 | ||||||||
| Unit | mM | ||||||||
| Comments | concentration dependent rates | ||||||||
| DATA_CREATED | |||||||||
| Datatype | rates | ||||||||
| Creation_date | 02/2009-05/2009 | ||||||||
| Status | row, normalised, .... | ||||||||
| Person_created | Tomas Fiedler | ||||||||
| DATA_FILE | |||||||||
| File_name | spy_ldh | ||||||||
| File_type | excel sheet | ||||||||
| Storage_location | SYSMOSEEK | ||||||||
| Storage_date | 2009-06-16T0:0:0+0000 | ||||||||
| Last_change_date | 2009-06-16T0:0:0+0000 | ||||||||
| TemplateID | Enzyme kinetics template | ||||||||
| PUBLICATION | |||||||||
| Title | 1 | ||||||||
| Authors | 1 | ||||||||
| Journal | 1 | ||||||||
| URL | 1 | ||||||||
| Year | 1 | ||||||||
| Volume | 1 | ||||||||
| Pages | 1 | ||||||||
| PubmedID | 1 | ||||||||
| UserID | 1 | ||||||||
| GroupID | 1 | ||||||||
| ProjectID | 1 | ||||||||
| ExperimentID | 1 | ||||||||
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| Pyruvate | LDH | LDH | Calculated affinity constant | ||||||
| Unit | mM | U/mg | U/mg | mM | |||||
| Experiment ID | Concentration | Average enzymatic rate | Standard deviation enzymatic rate | Km | |||||
| 1.01 | 0 | 0 | 0.41 | ||||||
| 1.02 | 0.05 | 0.508 | |||||||
| 1.03 | 0.1 | ||||||||
| 1.04 | 0.15 | 1.607 | |||||||
| 1.05 | 0.25 | 2.199 | |||||||
| 1.06 | 0.5 | ||||||||
| 1.07 | 0.8 | 5.16 | |||||||
| 1.08 | 1 | 6.091 | |||||||
| 1.09 | 2 | 6.091 | |||||||
| 1.1 | 2.5 | 6.091 | |||||||
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| NADH | LDH | LDH | Calculated affinity constant | ||||||
| Unit | mM | U/mg | U/mg | mM | |||||
| Experiment ID | Concentration | Average enzymatic rate | Standard deviation enzymatic rate | Km | |||||
| 2.01 | 0 | 0 | 0.062 | ||||||
| 2.0199999999999996 | 0.0125 | 0.384048 | |||||||
| 2.0299999999999994 | 0.025 | 1.300784 | |||||||
| 2.039999999999999 | 0.0375 | 2.078608 | |||||||
| 2.049999999999999 | 0.0625 | 2.927248 | |||||||
| 2.0599999999999987 | 0.125 | 4.498849 | |||||||
| 2.0699999999999985 | 0.175 | 5.436609 | |||||||
| 2.0799999999999983 | 0.25 | 5.980177 | |||||||
| 2.089999999999998 | 0.375 | 6.01431 | |||||||
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| Lactate | LDH | LDH | Calculated affinity constant | ||||||
| Unit | mM | U/mg | U/mg | mM | |||||
| Experiment ID | Concentration | Average enzymatic rate | Standard deviation enzymatic rate | Km | |||||
| 3.01 | 0 | 0 | 8.8 | ||||||
| 3.0199999999999996 | 5 | 0.15382 | |||||||
| 3.0299999999999994 | 10 | 0.48925 | |||||||
| 3.039999999999999 | 15 | 0.79145 | |||||||
| 3.049999999999999 | 20 | 0.79145 | |||||||
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| ATP | LDH | LDH | Calculated affinity constant | ||||||
| Unit | mM | U/mg | U/mg | mM | |||||
| Experiment ID | Concentration | Average enzymatic rate | Standard deviation enzymatic rate | Km | |||||
| 4.01 | 0 | 5.51115 | |||||||
| 4.02 | 0.5 | 6.216846 | |||||||
| 4.029999999999999 | 1 | 6.384869 | |||||||
| 4.039999999999999 | 1.25 | 6.284055 | |||||||
| 4.049999999999999 | 1.5 | 3.461271 | |||||||
| 4.059999999999999 | 2 | 3.192434 | |||||||
| 4.0699999999999985 | 2.5 | 5.208709 | |||||||
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| FBP | LDH | LDH | Calculated affinity constant | ||||||
| Unit | mM | U/mg | U/mg | mM | |||||
| Experiment ID | Concentration | Average enzymatic rate | Standard deviation enzymatic rate | Km | |||||
| 5.01 | 0 | 0.7697365 | 0.2 | ||||||
| 5.02 | 0.025 | 1.045654 | |||||||
| 5.029999999999999 | 0.25 | 8.669119 | |||||||
| 5.039999999999999 | 0.5 | 18.472296 | |||||||
| 5.049999999999999 | 1.25 | 13.491282 | |||||||
| 5.059999999999999 | 2.5 | 14.5376265 | |||||||
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| Phosphate | LDH | LDH | Calculated affinity constant | ||||||
| Unit | mM | U/mg | U/mg | mM | |||||
| Experiment ID | Concentration | Average enzymatic rate | Standard deviation enzymatic rate | Km | |||||
| 6.01 | 1 | 1.278415 | |||||||
| 6.02 | 20 | 20.582485 | |||||||
| 6.029999999999999 | 40 | 25.397849 | |||||||
| 6.039999999999999 | 50 | 25.52569 | |||||||
| 6.049999999999999 | 60 | 23.863751 | |||||||
| 6.059999999999999 | 70 | ||||||||
| 6.0699999999999985 | 80 | 16.662012 | |||||||
| 6.079999999999998 | 100 | 14.659161 | |||||||
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| Manganese | LDH | LDH | Calculated affinity constant | ||||||
| Unit | mM | U/mg | U/mg | mM | |||||
| Experiment ID | Concentration | Average enzymatic rate | Standard deviation enzymatic rate | Km | |||||
| 7.01 | 0 | 11.759079 | |||||||
| 7.02 | 0.25 | 4.671415 | |||||||
| 7.029999999999999 | 1.25 | 3.865999 | |||||||
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| Manganese | LDH | LDH | Calculated affinity constant | ||||||
| Unit | mM | U/mg | U/mg | mM | |||||
| Experiment ID | Concentration | Average enzymatic rate | Standard deviation enzymatic rate | Km | |||||
| 8.01 | 0 | 11.759079 | |||||||
| 8.02 | 0.25 | 2.094083 | |||||||
| 8.03 | 1.25 | 2.416249 | |||||||
| 8.04 | 2.5 | 2.255166 | |||||||
