Selected Cell
Cell:
Value:
Meta data
Organism_sample
AssayComposition
Results_calculated_Km Values
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| Item | Value | ||||||||
| Experimentator | Silvio Hering | ||||||||
| Raw data | |||||||||
| Reference | SYSMO-LAB | ||||||||
| Description | Heterologous Expression of LDHs from different lactic acid bacteria in Escherichia coli DH5α. Measurements on kinetics of L-LDHs from different latic acid bacterias at pH 6.0 and pH 7.0. | ||||||||
| Aim | Assessment of kinetic parameters of LDH to include in a catabolic model | ||||||||
| overexpression strain | E.coli DH5α | ||||||||
| Organism of Origin | S. pyogenes, E.faecalis, L.plantarum | ||||||||
| Vessel type | 384 wellplate | ||||||||
| Stirring rate | 0 | RPM | |||||||
| Temperature | 30 | degree celcius | |||||||
| Volume | 60 | µl | |||||||
| Gas flow | 0 | ml | |||||||
| pH | 7.0 and 6.0 | ||||||||
| SOP's used | 1 | ||||||||
| Explanations | Format | Data | |||||||
| TOPIC | |||||||||
| Titel | text | Heterologous Expression of LDHs from different lactic acid bacteria in Escherichia coli DH5α. Measurements on kinetics of L-LDHs from different latic acid bacterias at pH 6.0 and pH 7.0. | |||||||
| Comments | text | some comments | |||||||
| ASSAY | |||||||||
| Title | text | LDH kinetics | |||||||
| Assay_type | integer | enzymatic assay | |||||||
| Comments | purified enzyme | ||||||||
| EXPERIMENT | |||||||||
| Experiment_description | text | Forward reaction: Measurement at 380 nm-> Decrease of NADH Reverse direction: Measurement at 340 nm-> Increase of NADH | |||||||
| Experiment_type | text | Concentration series | |||||||
| Experimentalist | text | Silvio Hering | |||||||
| Begin_date | date | 2011-10-12T0:0:0+0000 | |||||||
| User | integer | Silvio Hering | |||||||
| Group | integer | SYSMOLAB | |||||||
| SOP | 1 | ||||||||
| Organism of Origin | |||||||||
| Organism | integer | S.pyogenes | L.plantarum | E.faeclis | |||||
| NCBI_ID | integer | 294934 | 220668 | 226185 | |||||
| Strain | text | M49 | WCFS1 | V583 | |||||
| Genotype | text | wild-type | wild-type | wild-type | |||||
| Phenotype | text | wild-type | wild-type | wild-type | |||||
| Experimental_conditions | |||||||||
| Item | text | pH | temperature | buffer | |||||
| Start_value | float/text | 7.0 | 30 | 1 | |||||
| End_value | float/text | 6.0 | 30 | 1 | |||||
| Unit | text | dimensionlesss | ºC | ||||||
| Comments | |||||||||
| FACTORS_STUDIED | |||||||||
| Item | text | Km | |||||||
| Start_value | float/text | 0 | |||||||
| End_value | float/text | 0 | |||||||
| Unit | text | mM | |||||||
| Comments | concentration dependent rates | ||||||||
| DATA_CREATED | |||||||||
| Datatype | text | rates | |||||||
| Creation_date | date | 08/2011-10/2011 | |||||||
| Status | text | row, normalised, .... | |||||||
| Person_created | text | Silvio Hering | |||||||
| DATA_FILE | |||||||||
| File_name | text | ldh_kinetic | |||||||
| File_type | text | excel sheet | |||||||
| Storage_location | text | SYSMOSEEK | |||||||
| Storage_date | date | 2012-03-30T0:0:0+0000 | |||||||
| Last_change_date | date | 2012-03-30T0:0:0+0000 | |||||||
| TemplateID | text | Enzyme kinetics template | |||||||
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| Treatment 1 | Treatment 2 (activator) | Organism/cell | Culture_description | ||||||||||||
| Sample_ID | Organism | Strain | Genotype | Strain comment | pH | Fructose-1.6-bisphophate | |||||||||
| Unit | mM | ||||||||||||||
| 1 | S.pyogenes | M49 | WT | E.coli DH5α IBA2_Sp_ldh | 7.0 | 3 | recombinant protein | L-lactate dehydrogenase/ E.C: 1.1.1.27/ MW: 35.28 kDa/ organism of origin: Spy49_0904c/length: 984 bp | |||||||
| 2 | S.pyogenes | M49 | WT | E.coli DH5α IBA2_Sp_ldh | 6.0 | 3 | recombinant protein | L-lactate dehydrogenase/ E.C: 1.1.1.27/ MW: 35.28 kDa/ organism of origin: Spy49_0904c/length: 984 bp | |||||||
| 3 | S.pyogenes | M49 | WT | E.coli DH5α IBA2_Sp_ldh | 7.0 | 3 | recombinant protein | L-lactate dehydrogenase/ E.C: 1.1.1.27/ MW: 35.28 kDa/ organism of origin: Spy49_0904c/length: 984 bp | |||||||
| 4 | S.pyogenes | M49 | WT | E.coli DH5α IBA2_Sp_ldh | 6.0 | 3 | recombinant protein | L-lactate dehydrogenase/ E.C: 1.1.1.27/ MW: 35.28 kDa/ organism of origin: Spy49_0904c/length: 984 bp | |||||||
| 5 | S.pyogenes | M49 | WT | E.coli DH5α IBA2_Sp_ldh | 7.0 | 3 | recombinant protein | L-lactate dehydrogenase/ E.C: 1.1.1.27/ MW: 35.28 kDa/ organism of origin: Spy49_0904c/length: 984 bp | |||||||
| 6 | S.pyogenes | M49 | WT | E.coli DH5α IBA2_Sp_ldh | 6.0 | 3 | recombinant protein | L-lactate dehydrogenase/ E.C: 1.1.1.27/ MW: 35.28 kDa/ organism of origin: Spy49_0904c/length: 984 bp | |||||||
| 7 | S.pyogenes | M49 | WT | E.coli DH5α IBA2_Sp_ldh | 7.0 | 3 | recombinant protein | L-lactate dehydrogenase/ E.C: 1.1.1.27/ MW: 35.28 kDa/ organism of origin: Spy49_0904c/length: 984 bp | |||||||
| 8 | S.pyogenes | M49 | WT | E.coli DH5α IBA2_Sp_ldh | 6.0 | 3 | recombinant protein | L-lactate dehydrogenase/ E.C: 1.1.1.27/ MW: 35.28 kDa/ organism of origin: Spy49_0904c/length: 984 bp | |||||||
| 9 | E.faecalis | V583 | WT | E.coli DH5α IBA2_Ef_ldh1 | 7.0 | 3 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 35.49 kDa/ organism of origin: EF0255/length: 984 bp | |||||||
| 10 | E.faecalis | V583 | WT | E.coli DH5α IBA2_Ef_ldh1 | 6.0 | 3 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 35.49 kDa/ organism of origin: EF0255/length: 984 bp | |||||||
| 11 | E.faecalis | V583 | WT | E.coli DH5α IBA2_Ef_ldh1 | 7.0 | 3 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 35.49 kDa/ organism of origin: EF0255/length: 984 bp | |||||||
| 12 | E.faecalis | V583 | WT | E.coli DH5α IBA2_Ef_ldh1 | 6.0 | 3 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 35.49 kDa/ organism of origin: EF0255/length: 984 bp | |||||||
| 13 | E.faecalis | V583 | WT | E.coli DH5α IBA2_Ef_ldh1 | 7.0 | 3 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 35.49 kDa/ organism of origin: EF0255/length: 984 bp | |||||||
| 14 | E.faecalis | V583 | WT | E.coli DH5α IBA2_Ef_ldh1 | 7.0 | 3 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 35.49 kDa/ organism of origin: EF0255/length: 984 bp | |||||||
| 15 | E.faecalis | V583 | WT | E.coli DH5α IBA2_Ef_ldh1 | 7.0 | 0 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 35.49 kDa/ organism of origin: EF0255/length: 984 bp | |||||||
| 16 | E.faecalis | V583 | WT | E.coli DH5α IBA2_Ef_ldh1 | 6.0 | 0 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 35.49 kDa/ organism of origin: EF0255/length: 984 bp | |||||||
| 17 | L.plantarum | WCFS1 | WT | E.coli DH5α IBA2_Lp_ldhL1 | 6.0 | 0 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 34.21 kDa/ organism of origin: lp_1101/length: 930 bp | |||||||
| 18 | L.plantarum | WCFS1 | WT | E.coli DH5α IBA2_Lp_ldhL1 | 6.0 | 0 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 34.21 kDa/ organism of origin: lp_1101/length: 930 bp | |||||||
| 19 | L.plantarum | WCFS1 | WT | E.coli DH5α IBA2_Lp_ldhL1 | 6.0 | 0 | recombinant protein | L-lactate dehydrogenase I / E.C: 1.1.1.27/ MW: 34.21 kDa/ organism of origin: lp_1101/length: 930 bp | |||||||
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| Identifer | Sample | Measurement | Organism | Enzyme | pH | Fructose-1.6-bisphophate | MES/KOH Buffer | MgCl2 | Pyruvate | NADH | Lactate | NAD+ |
| unit | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 | mM | mM | mM | mM | mM | mM | mM |
| 1 | Km Pyruvate | S.pyogenes M49 | L-LDH | 7.0 | 3 | 100 | 5 | 0.0 - 50.0 * | 4 | 0 | 0 | |
| 2 | Km Pyruvate | S.pyogenes M49 | L-LDH | 6.0 | 3 | 100 | 5 | 0.0 - 50.0 * | 4 | 0 | 0 | |
| 3 | Km NADH | S.pyogenes M49 | L-LDH | 7.0 | 3 | 100 | 5 | 50 | 0.0 - 4.0 * | 0 | 0 | |
| 4 | Km NADH | S.pyogenes M49 | L-LDH | 6.0 | 3 | 100 | 5 | 50 | 0.0 - 4.0 * | 0 | 0 | |
| 5 | Km Lactate | S.pyogenes M49 | L-LDH | 7.0 | 3 | 100 | 5 | 0 | 0 | 0.0 - 500 * | 10 | |
| 6 | Km Lactate | S.pyogenes M49 | L-LDH | 6.0 | 3 | 100 | 5 | 0 | 0 | 0.0 - 500 * | 10 | |
| 7 | Km NAD+ | S.pyogenes M49 | L-LDH | 7.0 | 3 | 100 | 5 | 0 | 0 | 500 | 0.0 - 10.0 * | |
| 8 | Km NAD+ | S.pyogenes M49 | L-LDH | 6.0 | 3 | 100 | 5 | 0 | 0 | 500 | 0.0 - 10.0 * | |
| 9 | Km Pyruvate | E.faecalis V583 | L-LDHLI | 7.0 | 3 | 100 | 5 | 0.0 - 50.0 * | 4 | 0 | 0 | |
| 10 | Km Pyruvate | E.faecalis V583 | L-LDHLI | 6.0 | 3 | 100 | 5 | 0.0 - 50.0 * | 4 | 0 | 0 | |
| 11 | Km NADH | E.faecalis V583 | L-LDHLI | 7.0 | 3 | 100 | 5 | 50 | 0.0 - 4.0 * | 0 | 0 | |
| 12 | Km NADH | E.faecalis V583 | L-LDHLI | 6.0 | 3 | 100 | 5 | 50 | 0.0 - 4.0 * | 0 | 0 | |
| 13 | Km Lactate | E.faecalis V583 | L-LDHLI | 7.0 | 3 | 100 | 5 | 0 | 0 | 0.0 - 500 * | 10 | |
| 14 | Km NAD+ | E.faecalis V583 | L-LDHLI | 7.0 | 3 | 100 | 5 | 0 | 0 | 500 | 0.0 - 10.0 * | |
| 15 | Km NADH | E.faecalis V583 | L-LDHLI | 7.0 | 0 | 100 | 5 | 50 | 0.0 - 4.0 * | 0 | 0 | |
| 16 | Km NADH | E.faecalis V583 | L-LDHLI | 6.0 | 0 | 100 | 5 | 50 | 0.0 - 4.0 * | 0 | 0 | |
| 17 | Km Pyruvate | L.plantarum WCFS1 | L-LDHLI | 6.0 | 0 | 100 | 5 | 0.0 - 50.0 * | 4 | 0 | 0 | |
| 18 | Km NADH | L.plantarum WCFS1 | L-LDHLI | 6.0 | 0 | 100 | 5 | 50 | 0.0 - 4.0 * | 0 | 0 | |
| 19 | Km Lactate | L.plantarum WCFS1 | L-LDHLI | 6.0 | 0 | 100 | 5 | 0 | 0 | 0.0 - 500 * | 10 | |
| * varied for the measurements (concentration dependent rates) | ||||||||||||
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| Identifer | Sample | Organism | Enzyme | Subtrate | pH | Km value | standard deviation Km | commentary | |||||
| unit | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 | mM | % | ||||||
| 1 | S.pyogenes M49 | L-LDH | Pyruvate | 7.0 | 1.69 | ± 5.55 | in presence of fructose-1.6-bisphosphate | ||||||
| 2 | S.pyogenes M49 | L-LDH | Pyruvate | 6.0 | 1.76 | ± 7.63 | in presence of fructose-1.6-bisphosphate | ||||||
| 3 | S.pyogenes M49 | L-LDH | NADH | 7.0 | 0.56 | ± 6.22 | in presence of fructose-1.6-bisphosphate | ||||||
| 4 | S.pyogenes M49 | L-LDH | NADH | 6.0 | 0.63 | ± 5.74 | in presence of fructose-1.6-bisphosphate | ||||||
| 5 | S.pyogenes M49 | L-LDH | Lactate | 7.0 | 98.04 | ± 5.22 | in presence of fructose-1.6-bisphosphate | ||||||
| 6 | S.pyogenes M49 | L-LDH | Lactate | 6.0 | 112 | ± 7.60 | in presence of fructose-1.6-bisphosphate | ||||||
| 7 | S.pyogenes M49 | L-LDH | NAD+ | 7.0 | 2.27 | ± 3.84 | in presence of fructose-1.6-bisphosphate | ||||||
| 8 | S.pyogenes M49 | L-LDH | NAD+ | 6.0 | 2.35 | ± 7.02 | in presence of fructose-1.6-bisphosphate | ||||||
| 9 | E.faecalis V583 | L-LDHLI | Pyruvate | 7.0 | 2.56 | ± 6.00 | in presence of fructose-1.6-bisphosphate | ||||||
| 10 | E.faecalis V583 | L-LDHLI | Pyruvate | 6.0 | 2.68 | ± 6.60 | in presence of fructose-1.6-bisphosphate | ||||||
| 11 | E.faecalis V583 | L-LDHLI | NADH | 7.0 | 0.56 | ± 5.30 | in presence of fructose-1.6-bisphosphate | ||||||
| 12 | E.faecalis V583 | L-LDHLI | NADH | 6.0 | 0.53 | ± 7.40 | in presence of fructose-1.6-bisphosphate | ||||||
| 13 | E.faecalis V583 | L-LDHLI | Lactate | 7.0 | 96.35 | ± 6.18 | in presence of fructose-1.6-bisphosphate | ||||||
| 14 | E.faecalis V583 | L-LDHLI | NAD+ | 7.0 | 2.68 | ± 9.43 | in presence of fructose-1.6-bisphosphate | ||||||
| 15 | E.faecalis V583 | L-LDHLI | NADH | 7.0 | 1.58 | ± 4.1 | in absence of fructose-1.6-bisphosphate | ||||||
| 16 | E.faecalis V583 | L-LDHLI | NADH | 6.0 | 1.56 | ± 4.9 | in absence of fructose-1.6-bisphosphate | ||||||
| 17 | L.plantarum WCFS1 | L-LDHLI | Pyruvate | 6.0 | 4.25 | ± 10.2 | in absence of fructose-1.6-bisphosphate | ||||||
| 18 | L.plantarum WCFS1 | L-LDHLI | NADH | 6.0 | 0.37 | ± 6.38 | in absence of fructose-1.6-bisphosphate | ||||||
| 19 | L.plantarum WCFS1 | L-LDHLI | Lactate | 6.0 | 103.05 | ± 7.72 | in absence of fructose-1.6-bisphosphate | ||||||
