Selected Cell
Cell:
Value:
Introduction
ResultsTable
Bradford assays
LS_ATA WT
SingleMutant V37A
SingleMutant S248A
SingleMutant T249A
Double mutants
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Photometric determination of activity | |||||||||||||||||||
Methodology | |||||||||||||||||||
The U ml-1 of each enzyme was determined spectrophotometrically based on the conversion | |||||||||||||||||||
of 1-phenylethylamine, 1-phenylpropylamine and 1-phenyl butylamine to their corresponding ketones. | |||||||||||||||||||
Deamination of the substrates leads to an increase in UV absorbtion, | |||||||||||||||||||
which can be detected spectrophotometrically at 245 nm (Schätzle et al. 2009). | |||||||||||||||||||
One unit of activity was defined as the amount of enzyme that produced 1 µmol of the ketone from its corresponding amine per minute. | |||||||||||||||||||
Assay Conditions | |||||||||||||||||||
The reaction mixture contained enantiopure 1.0 mM of the (R)- enantiomer of the amine and 2.0 mM and sodium pyruvate as keto-donor. | |||||||||||||||||||
The amines were diluted in DMSO, 5% final concetration. | |||||||||||||||||||
All enzymes were assayed in purified form. | |||||||||||||||||||
(Ni-NTA 5 mL Protino column were used for purification and HiTrapTM 15 mL for desalting. Purified enzyme solutions were stored at -20 °C after flash freezing) | |||||||||||||||||||
Enzyme concetrations were determined using the Bradford assay. | |||||||||||||||||||
The pH of the buffer was adjusted at 30 °C. | |||||||||||||||||||
All buffer and reagent solutions were brought to room temperature for the assay. | |||||||||||||||||||
The assay componets were pipetted in the following order: | |||||||||||||||||||
5μl of enzyme | |||||||||||||||||||
10 μl of amine stock solution | |||||||||||||||||||
85 μl HEPES buffer pH= 8 | |||||||||||||||||||
100 μl of pyruvate 2.0 mM (HEPES buffer 50mM) solution was added last with a multipipet for the initiation of the reaction. | |||||||||||||||||||
Photometer parameters | |||||||||||||||||||
Set temperature | 30 °C | ||||||||||||||||||
Plate template | Greiner UV 96-well plate | ||||||||||||||||||
Wavelength [nm] | 245 | ||||||||||||||||||
Kinetic interval | 00:00:10 | ||||||||||||||||||
Kinetic total time | 00:05:00 | ||||||||||||||||||
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Kinetic assay LS_ATA and single mutants _ pH= 8.0_ R-PEA, R-PPA, R-PBA | |||||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | |||||||||||||||
(R)- phenylethylamine | (R)- phenylpropylamine | (R)- phenylbutylamine | |||||||||||||
U /ml | STDEV | (mg/ml) | U/mg | STDEV | U /ml | STDEV | (mg/ml) | U/mg | STDEV | U /ml | STDEV | (mg/ml) | U/mg | STDEV | |
GamPro WT | 1.614299725775136 | 0.3177919691049235 | 5.893719806763285 | 0.2739016747831583 | 0.05392044065960587 | _ | _ | ||||||||
GamPro V37A | 0.06258676114552808 | 0.023678315611166106 | 0.671450084152715 | 0.09321133859787158 | 0.015898807009712027 | 0.030433006929736474 | 0.017788911663543325 | 0.671450084152715 | 0.04532430280076452 | 0.02649327490365971 | _ | ||||
GamPro S248A | _ | _ | _ | ||||||||||||
GamPro T249A | 0.08166040172631177 | 0.021320818256139077 | 0.5926122774382141 | 0.13779735053637274 | 0.03597768569410375 | _ | _ | ||||||||
Kinetic assay_ LS_ATA double mutants V37A/S248A and V37A/T249A pH= 8.0_ R-PEA, R-PPA, R-PBA | |||||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | |||||||||||||||
(R)- phenylethylamine | (R)- phenylpropylamine | (R)- phenylbutylamine | |||||||||||||
U /ml | STDEV | (mg/ml) | U/mg | STDEV | U /ml | STDEV | (mg/ml) | U/mg | STDEV | U /ml | STDEV | (mg/ml) | U/mg | STDEV | |
GamPro V37A/S248A | _ | _ | _ | ||||||||||||
GamPro V37A/T249A | _ | 0.0011504596032897907 | 4.6548115461454085E-4 | 0.64171331857831 | 0.0017927937133026747 | 7.25372438343333E-4 | _ | ||||||||
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Bradford | |||||||||||||||
Endpoint | |||||||||||||||
Wavelength: 595 nm | |||||||||||||||
Set temperature 30 °C | |||||||||||||||
96 well plate | |||||||||||||||
Shake yes | |||||||||||||||
200 μl of bradford reagent and 15 μl sample | |||||||||||||||
LS_ATA double mutants V371A/S248A and V37A/T249A from 500 mL culture after purification | |||||||||||||||
Standart curve | |||||||||||||||
Well | Absorbance BSA | Absorbance BSA av. | STDEV | Absorbance BSA av. | BSA (mg/ml) | ||||||||||
A 1 | 0.4146 | 0.41673333333333334 | 0.009285652014443209 | 0.41673333333333334 | 0 | ||||||||||
A 2 | 0.4269 | 0.5462666666666666 | 0.1 | ||||||||||||
A 3 | 0.4087 | 0.6434333333333333 | 0.2 | ||||||||||||
B 1 | 0.5423 | 0.5462666666666666 | 0.0048686069191641685 | 0.7376333333333333 | 0.3 | ||||||||||
B 2 | 0.5517 | 0.8254999999999999 | 0.4 | ||||||||||||
B 3 | 0.5448 | 0.9317666666666667 | 0.6 | ||||||||||||
C 1 | 0.6342 | 0.6434333333333333 | 0.02223743090676917 | 1.0473666666666666 | 0.8 | ||||||||||
C 2 | 0.6688 | ||||||||||||||
C 3 | 0.6273 | ||||||||||||||
D 1 | 0.714 | 0.7376333333333333 | 0.02782486897243783 | ||||||||||||
D 2 | 0.7683 | ||||||||||||||
D 3 | 0.7306 | ||||||||||||||
E 1 | 0.833 | 0.8254999999999999 | 0.027916124372842306 | ||||||||||||
E 2 | 0.8489 | ||||||||||||||
E 3 | 0.7946 | ||||||||||||||
F 1 | 0.9174 | 0.9317666666666667 | 0.029683384802500743 | ||||||||||||
F 2 | 0.9659 | ||||||||||||||
F 3 | 0.912 | ||||||||||||||
G 1 | 1.0632 | 1.0473666666666666 | 0.023471756076896607 | ||||||||||||
G 2 | 1.0585 | ||||||||||||||
G 3 | 1.0204 | ||||||||||||||
Results | (y-0,4722)/0,7681 | ||||||||||||||
Sample | Absorbance (AU) | (mg/ml) | Dilution Factor | (mg/ml) * DF | Concetration (mg/ml) | STDEV | |||||||||
V37A/S248A | 0.9057 | 0.5643796380679598 | 1 | 0.5602786095560472 | 0.005799730141225357 | ||||||||||
0.8994 | 0.5561775810441348 | 1 | |||||||||||||
V37A/T249A | 0.9691 | 0.6469209738315322 | 1 | 0.64171331857831 | 0.0073647366872703285 | ||||||||||
0.9611 | 0.6365056633250878 | 1 | |||||||||||||
Bradford | |||||||||||||||
Endpoint | |||||||||||||||
Wavelength: 595 nm | |||||||||||||||
Set temperature 30 °C | |||||||||||||||
96 well plate | |||||||||||||||
Shake Υes, Shake duration (00:00:25) | |||||||||||||||
200 μl of bradford reagent and 10 μl sample | |||||||||||||||
LS_ATA single mutants: V371, S248A, T249A from a 500 mL culture after purification. | |||||||||||||||
Standart curve | |||||||||||||||
Well | Absorbance BSA | Absorbance BSA av. | STDEV | Absorbance BSA av. | BSA (mg/ml) | ||||||||||
A 1 | 0.3672 | 0.36810000000000004 | 0.01227477087362531 | 0.36810000000000004 | 0 | ||||||||||
A 2 | 0.3563 | 0.428 | 0.1 | ||||||||||||
A 3 | 0.3808 | 0.4605666666666666 | 0.2 | ||||||||||||
B 1 | 0.4234 | 0.428 | 0.007709085548883214 | 0.4829666666666667 | 0.3 | ||||||||||
B 2 | 0.4369 | 0.5148666666666667 | 0.4 | ||||||||||||
B 3 | 0.4237 | 0.6097333333333333 | 0.6 | ||||||||||||
C 1 | 0.4683 | 0.4605666666666666 | 0.008730024818597783 | ||||||||||||
C 2 | 0.4511 | ||||||||||||||
C 3 | 0.4623 | ||||||||||||||
D 1 | 0.4795 | 0.4829666666666667 | 0.022104826019069563 | ||||||||||||
D 2 | 0.5066 | ||||||||||||||
D 3 | 0.4628 | ||||||||||||||
E 1 | 0.5277 | 0.5148666666666667 | 0.02068872478751006 | ||||||||||||
E 2 | 0.491 | ||||||||||||||
E 3 | 0.5259 | ||||||||||||||
F 1 | 0.6003 | 0.6097333333333333 | 0.030947751668470733 | ||||||||||||
F 2 | 0.6443 | ||||||||||||||
F 3 | 0.5846 | ||||||||||||||
G 1 | 0.5609 | 0.6262 | 0.0728661100924154 | ||||||||||||
G 2 | 0.7048 | ||||||||||||||
G 3 | 0.6129 | ||||||||||||||
Results | (y-0,377)/0,3763 | ||||||||||||||
Sample | Absorbance (AU) | (mg/ml) | Dilution Factor | (mg/ml) * DF | Concetration (mg/ml) | STDEV | |||||||||
LS_ATA WT | 5,89 ± 0,5 mg/mL | ||||||||||||||
(previously determined using the same assay) | |||||||||||||||
V37A | 0.666 | 0.7680042519266543 | 1 | 0.7680042519266543 | 0.671450084152715 | 0.08413406416441027 | |||||||||
0.615 | 0.6324740898219505 | 1 | 0.6324740898219505 | ||||||||||||
0.608 | 0.6138719107095402 | 1 | 0.6138719107095402 | ||||||||||||
S248A | 0.554 | 0.4703693861280894 | 1 | 0.4703693861280894 | 0.48188502081672424 | 0.0684949895375076 | |||||||||
0.535 | 0.4198777571086899 | 1 | 0.4198777571086899 | ||||||||||||
0.586 | 0.5554079192133934 | 1 | 0.5554079192133934 | ||||||||||||
T249A | 0.612 | 0.6245017273452033 | 1 | 0.6245017273452033 | 0.5926122774382141 | 0.04422885191892971 | |||||||||
0.581 | 0.5421206484188147 | 1 | 0.5421206484188147 |
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Kinetic assay_ LS_ATA WT | |||||||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | mM/min | ||||||||||||||||
R-PEA | *60 | (ΔΑbs/min) / (9,15 Αbs/mM) | *DF | 40 | *αραίωση πηγαδιού | μmol/(ml * min) | (Bradford) | μmol/ (mg * min) | |||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ΔΑbs/min | mmol/ (L * min) | DF | mmol/ (L * min) | αραιωση πηγαδιού | mmol/ (L * min) | U /ml | STDEV | (mg/ml) | U/mg | STDEV | |
A 1 | blank | 6.838827838827854E-5 | 0.9785394580384066 | 5.8772893772893795E-5 | |||||||||||||
B 1 | blank | 4.9157509157509046E-5 | 0.9861384815792107 | 5.8772893772893795E-5 | |||||||||||||
C 1 | LS_ATA WT (1:5) | 0.0012380708180708183 | 0.9994032900058445 | 5.8772893772893795E-5 | 0.0011792979242979246 | 0.07075787545787547 | 0.00773310114293721 | 5 | 0.03866550571468605 | 40 | 1.546620228587442 | 1.614299725775136 | 0.3177919691049235 | 5.893719806763285 | 0.2739016747831583 | 0.05392044065960587 | |
D 1 | LS_ATA WT (1:5) | 0.0015536385836385832 | 0.9998518577980429 | 5.8772893772893795E-5 | 0.0014948656898656895 | 0.08969194139194137 | 0.00980239796633239 | 5 | 0.04901198983166195 | 40 | 1.960479593266478 | ||||||
E 1 | LS_ATA WT (1:5) | 0.0010773199023199028 | 0.9997520184120487 | 5.8772893772893795E-5 | 0.001018547008547009 | 0.06111282051282055 | 0.006678996777357437 | 5 | 0.03339498388678718 | 40 | 1.3357993554714873 | ||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | |||||||||||||||||
R-PPA | |||||||||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ||||||||||||
A 2 | blank (R)-PPA | -2.7100122100122095E-5 | 0.8757066340794218 | -1.842301418388365E-5 | |||||||||||||
B 2 | blank (R)-PPA | -9.745906267645204E-6 | 0.2731558553255219 | -1.842301418388365E-5 | |||||||||||||
C 2 | LS_ATA WT (1:5) | -7.25274725274725E-5 | 0.9229747993249092 | -1.842301418388365E-5 | -7.25274725274725E-5 | ||||||||||||
D 2 | LS_ATA WT (1:5) | -2.0152625152625318E-5 | 0.8051492374902609 | -1.842301418388365E-5 | -2.0152625152625318E-5 | ||||||||||||
E 2 | LS_ATA WT (1:5) | -3.155677655677657E-5 | 0.8450943321986674 | -1.842301418388365E-5 | -3.155677655677657E-5 | ||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | |||||||||||||||||
R- PBA | |||||||||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ||||||||||||
A 3 | blank (R)-PBA | -3.072588522588522E-4 | 0.9553380902164618 | -2.817765567765568E-4 | |||||||||||||
B 3 | blank (R)-PBA | -2.562942612942614E-4 | 0.9517267438066959 | -2.817765567765568E-4 | |||||||||||||
C 3 | LS_ATA WT (1:5) | -4.175396825396834E-4 | 0.9576819182329621 | -2.817765567765568E-4 | -4.175396825396834E-4 | ||||||||||||
D 3 | LS_ATA WT (1:5) | -2.713614163614164E-4 | 0.9674420524366556 | -2.817765567765568E-4 | -2.713614163614164E-4 | ||||||||||||
E 3 | LS_ATA WT (1:5) | -4.932031497092351E-4 | 0.9670394024095663 | -2.817765567765568E-4 | -4.932031497092351E-4 | ||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | |||||||||||||||||
R- PBA | |||||||||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ||||||||||||
A 4 | blank (R)-PBA | -1.5556419938063878E-4 | 0.8597220520967417 | -1.5556419938063878E-4 | |||||||||||||
B 4 | blank (R)-PBA | -9.928190894789688E-6 | 0.011592267255954644 | -1.5556419938063878E-4 | |||||||||||||
C 4 | LS_ATA WT (1:5) | -1.793484578929603E-4 | 0.9492100476360545 | -1.5556419938063878E-4 | -1.793484578929603E-4 | ||||||||||||
D 4 | LS_ATA WT (1:5) | -8.906678451410196E-5 | 0.6573964551575117 | -1.5556419938063878E-4 | -8.906678451410196E-5 | ||||||||||||
E 4 | LS_ATA WT (1:5) | -6.0532320729967035E-5 | 0.8998765675319981 | -1.5556419938063878E-4 | -6.0532320729967035E-5 | ||||||||||||
Measurements | |||||||||||||||||
WT R-PEA | WT_R-PPA_R-PBA | ||||||||||||||||
Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | ||||||||
Α1 blank (R) | 0.6701 | 0 | 6.838827838827854E-5 | 0.9785394580384066 | A 2 | 0.7775 | 0.02 | -2.7100122100122095E-5 | 0.8757066340794218 | ||||||||
A 1 | 0.67 | 10.46 | A 2 | 0.7897 | 10.49 | ||||||||||||
A 1 | 0.6708 | 20.46 | A 2 | 0.7877 | 20.49 | ||||||||||||
A 1 | 0.6717 | 30.46 | A 2 | 0.7887 | 30.49 | ||||||||||||
A 1 | 0.6727 | 40.46 | A 2 | 0.7883 | 40.49 | ||||||||||||
A 1 | 0.6736 | 50.46 | A 2 | 0.7877 | 50.49 | ||||||||||||
A 1 | 0.6748 | 60.46 | A 2 | 0.7867 | 60.49 | ||||||||||||
A 1 | 0.6757 | 70.46 | A 2 | 0.7864 | 70.49 | ||||||||||||
A 1 | 0.6769 | 80.46 | A 2 | 0.786 | 80.49 | ||||||||||||
A 1 | 0.6779 | 90.46 | A 2 | 0.7857 | 90.49 | ||||||||||||
A 1 | 0.6788 | 100.46 | A 2 | 0.7852 | 100.49 | ||||||||||||
A 1 | 0.6796 | 110.46 | A 2 | 0.784 | 110.49 | ||||||||||||
A 1 | 0.6802 | 120.46 | A 2 | 0.7839 | 120.49 | ||||||||||||
A 1 | 0.6816 | 130.46 | A 2 | 0.7838 | 130.49 | ||||||||||||
A 1 | 0.6817 | 140.46 | A 2 | 0.7839 | 140.49 | ||||||||||||
A 1 | 0.6828 | 150.46 | A 2 | 0.7834 | 150.49 | ||||||||||||
A 1 | 0.6833 | 160.46 | A 2 | 0.783 | 160.49 | ||||||||||||
A 1 | 0.6837 | 170.46 | A 2 | 0.7815 | 170.49 | ||||||||||||
A 1 | 0.6846 | 180.46 | A 2 | 0.7825 | 180.49 | ||||||||||||
A 1 | 0.6855 | 190.46 | A 2 | 0.7822 | 190.49 | ||||||||||||
A 1 | 0.6859 | 200.46 | A 2 | 0.7815 | 200.49 | ||||||||||||
A 1 | 0.6863 | 210.46 | A 2 | 0.7817 | 210.49 | ||||||||||||
A 1 | 0.6859 | 220.46 | A 2 | 0.7833 | 220.49 | ||||||||||||
A 1 | 0.6878 | 230.46 | A 2 | 0.7817 | 230.49 | ||||||||||||
A 1 | 0.6878 | 240.46 | A 2 | 0.7819 | 240.49 | ||||||||||||
A 1 | 0.6884 | 250.46 | A 2 | 0.7825 | 250.49 | ||||||||||||
A 1 | 0.6892 | 260.46 | A 2 | 0.7812 | 260.49 | ||||||||||||
A 1 | 0.6898 | 270.46 | A 2 | 0.78 | 270.49 | ||||||||||||
A 1 | 0.6903 | 280.46 | A 2 | 0.781 | 280.49 | ||||||||||||
A 1 | 0.6895 | 290.46 | A 2 | 0.7801 | 290.49 | ||||||||||||
A 1 | 0.6915 | 300.46 | A 2 | 0.7807 | 300.49 | ||||||||||||
B1 blank (R) | 0.706 | 0.42 | 4.9157509157509046E-5 | 0.9861384815792107 | A 3 | 2.5264 | 0.04 | -3.072588522588522E-4 | 0.9553380902164618 | ||||||||
B 1 | 0.702 | 10.89 | A 3 | 2.5472 | 10.51 | ||||||||||||
B 1 | 0.7041 | 20.89 | A 3 | 2.5487 | 20.51 | ||||||||||||
B 1 | 0.7046 | 30.89 | A 3 | 2.5482 | 30.51 | ||||||||||||
B 1 | 0.7054 | 40.89 | A 3 | 2.5448 | 40.51 | ||||||||||||
B 1 | 0.7061 | 50.89 | A 3 | 2.5453 | 50.51 | ||||||||||||
B 1 | 0.7069 | 60.89 | A 3 | 2.5419 | 60.51 | ||||||||||||
B 1 | 0.7076 | 70.89 | A 3 | 2.5417 | 70.51 | ||||||||||||
B 1 | 0.7083 | 80.89 | A 3 | 2.5321 | 80.51 | ||||||||||||
B 1 | 0.7092 | 90.89 | A 3 | 2.5288 | 90.51 | ||||||||||||
B 1 | 0.7096 | 100.89 | A 3 | 2.5253 | 100.51 | ||||||||||||
B 1 | 0.71 | 110.89 | A 3 | 2.5216 | 110.51 | ||||||||||||
B 1 | 0.7102 | 120.89 | A 3 | 2.5201 | 120.51 | ||||||||||||
B 1 | 0.7106 | 130.89 | A 3 | 2.523 | 130.51 | ||||||||||||
B 1 | 0.7113 | 140.89 | A 3 | 2.5184 | 140.51 | ||||||||||||
B 1 | 0.7117 | 150.89 | A 3 | 2.5271 | 150.51 | ||||||||||||
B 1 | 0.7118 | 160.89 | A 3 | 2.5193 | 160.51 | ||||||||||||
B 1 | 0.7124 | 170.89 | A 3 | 2.5122 | 170.51 | ||||||||||||
B 1 | 0.713 | 180.89 | A 3 | 2.502 | 180.51 | ||||||||||||
B 1 | 0.7146 | 190.89 | A 3 | 2.4965 | 190.51 | ||||||||||||
B 1 | 0.7143 | 200.89 | A 3 | 2.4948 | 200.51 | ||||||||||||
B 1 | 0.7148 | 210.89 | A 3 | 2.4928 | 210.51 | ||||||||||||
B 1 | 0.7152 | 220.89 | A 3 | 2.496 | 220.51 | ||||||||||||
B 1 | 0.7159 | 230.89 | A 3 | 2.4935 | 230.51 | ||||||||||||
B 1 | 0.7161 | 240.89 | A 3 | 2.4884 | 240.51 | ||||||||||||
B 1 | 0.7167 | 250.89 | A 3 | 2.4845 | 250.51 |
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Kinetic assay_single mutant V37A _ pH= 8.0_ R-PEA, R-PPA, R-PBA | ||||||||||||||||
6,36 Abs/mM butyrophenone | ||||||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | mM/min | |||||||||||||||
R-PEA | *60 | (ΔΑbs/min) / (9,15 Αbs/mM) | *DF | 40 | *αραίωση πηγαδιού | μmol/(ml * min) | (Bradford) | μmol/ (mg * min) | ||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ΔΑbs/min | mmol/ (L * min) | DF | mmol/ (L * min) | αραιωση πηγαδιού | mmol/ (L * min) | U /ml | STDEV | (mg/ml) | U/mg | STDEV |
A 5 | blank (R)-PEA | -1.2923755913962563E-5 | 0.869912148780359 | -3.6414868187209005E-6 | ||||||||||||
B 5 | blank (R)-PEA | 5.6407822765207626E-6 | 0.7801197324348885 | -3.6414868187209005E-6 | ||||||||||||
C 5 | V37 (1:5) | 5.350242872480166E-5 | 0.9834772466989931 | -3.6414868187209005E-6 | 5.350242872480166E-5 | 0.0032101457234881 | 3.5121944458294305E-4 | 5 | 0.0017560972229147152 | 40 | 0.07024388891658861 | 0.06258676114552808 | 0.023678315611166106 | 0.671450084152715 | 0.09321133859787158 | 0.015898807009712027 |
D 5 | V37 (1:5) | 2.7440873321900363E-5 | 0.912213868146465 | -3.6414868187209005E-6 | 2.7440873321900363E-5 | 0.0016464523993140218 | 1.8013702399496956E-4 | 5 | 9.006851199748478E-4 | 40 | 0.03602740479899391 | |||||
E 5 | V37 (1:5) | 6.206744717082964E-5 | 0.9883631170621836 | -3.6414868187209005E-6 | 6.206744717082964E-5 | 0.003724046830249778 | 4.0744494860500853E-4 | 5 | 0.0020372247430250428 | 40 | 0.08148898972100171 | |||||
HEPES 50 mM pH 8.0 0,05 mM PLP | ||||||||||||||||
R-PPA | ||||||||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ΔΑbs/min | mmol/ (L * min) | DF | mmol/ (L * min) | αραιωση πηγαδιού | mmol/ (L * min) | U /ml | STDEV | (mg/ml) | U/mg | STDEV |
A 6 | blank (R)-PPA | -2.6617848793936036E-5 | 0.9543872734766372 | -2.5125186745895983E-5 | ||||||||||||
B 6 | blank (R)-PPA | -2.3632524697855934E-5 | 0.9717508669540801 | -2.5125186745895983E-5 | ||||||||||||
C 6 | V37 (1:5) | 2.279618350838691E-5 | 0.9582977627409738 | -2.5125186745895983E-5 | 2.279618350838691E-5 | 0.0013677710105032144 | 2.1505833498478214E-4 | 5 | 0.0010752916749239107 | 40 | 0.04301166699695643 | 0.030433006929736474 | 0.017788911663543325 | 0.671450084152715 | 0.04532430280076452 | 0.02649327490365971 |
D 6 | V37 (1:5) | -4.156300097517312E-5 | 0.7386450357256938 | -2.5125186745895983E-5 | -4.156300097517312E-5 | |||||||||||
E 6 | V37 (1:5) | 9.462803837133751E-6 | 0.7554149768896209 | -2.5125186745895983E-5 | 9.462803837133751E-6 | 5.677682302280251E-4 | 8.927173431258256E-5 | 5 | 4.463586715629128E-4 | 40 | 0.017854346862516513 | |||||
HEPES 50 mM pH 8.0 0,05 mM PLP | ||||||||||||||||
R-PBA | ||||||||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | |||||||||||
A 7 | blank (R)-PBA | -2.47258852258852E-4 | 0.9788259027428547 | -2.3340354090354068E-4 | ||||||||||||
B 7 | blank (R)-PBA | -2.1954822954822938E-4 | 0.9726877232195452 | -2.3340354090354068E-4 | ||||||||||||
C 7 | V37 (1:5) | -1.9185592185592146E-4 | 0.9940423055765812 | -2.3340354090354068E-4 | -1.9185592185592146E-4 | |||||||||||
D 7 | V37 (1:5) | -1.68687423687424E-4 | 0.7941246371645544 | -2.3340354090354068E-4 | -1.68687423687424E-4 | |||||||||||
E 7 | V37 (1:5) | -2.2652625152625174E-4 | 0.988125385218469 | -2.3340354090354068E-4 | -2.2652625152625174E-4 | |||||||||||
Measurements | ||||||||||||||||
V37_1;5_PEA | V37_1;5_PPA | V37_1;5_PBA | ||||||||||||||
Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | ||
A 5 | 0.7075 | 0 | -1.2923755913962563E-5 | 0.869912148780359 | A 6 | 0.695 | 0 | -2.6617848793936036E-5 | 0.9543872734766372 | A 7 | 1.6438 | 0 | -2.47258852258852E-4 | 0.9788259027428547 | ||
A 5 | 0.7054 | 10.39 | A 6 | 0.6956 | 10.38 | A 7 | 1.6552 | 10.36 | ||||||||
A 5 | 0.7034 | 20.39 | A 6 | 0.7001 | 20.38 | A 7 | 1.6501 | 20.36 | ||||||||
A 5 | 0.7035 | 30.4 | A 6 | 0.7003 | 30.38 | A 7 | 1.6575 | 30.36 | ||||||||
A 5 | 0.7023 | 40.39 | A 6 | 0.7013 | 40.38 | A 7 | 1.6492 | 40.36 | ||||||||
A 5 | 0.7028 | 50.39 | A 6 | 0.7008 | 50.38 | A 7 | 1.6475 | 50.36 | ||||||||
A 5 | 0.7033 | 60.4 | A 6 | 0.7005 | 60.38 | A 7 | 1.6476 | 60.36 | ||||||||
A 5 | 0.7022 | 70.39 | A 6 | 0.7 | 70.38 | A 7 | 1.6437 | 70.36 | ||||||||
A 5 | 0.7015 | 80.39 | A 6 | 0.6998 | 80.38 | A 7 | 1.6453 | 80.36 | ||||||||
A 5 | 0.7027 | 90.4 | A 6 | 0.6997 | 90.39 | A 7 | 1.6375 | 90.36 | ||||||||
A 5 | 0.7014 | 100.39 | A 6 | 0.6987 | 100.38 | A 7 | 1.6342 | 100.36 | ||||||||
A 5 | 0.7009 | 110.39 | A 6 | 0.6985 | 110.38 | A 7 | 1.6308 | 110.36 | ||||||||
A 5 | 0.7012 | 120.4 | A 6 | 0.6981 | 120.38 | A 7 | 1.6282 | 120.36 | ||||||||
A 5 | 0.7013 | 130.4 | A 6 | 0.6975 | 130.38 | A 7 | 1.6286 | 130.36 | ||||||||
A 5 | 0.701 | 140.4 | A 6 | 0.6977 | 140.38 | A 7 | 1.6244 | 140.36 | ||||||||
A 5 | 0.7009 | 150.4 | A 6 | 0.6974 | 150.38 | A 7 | 1.6204 | 150.36 | ||||||||
A 5 | 0.7013 | 160.4 | A 6 | 0.697 | 160.38 | A 7 | 1.6172 | 160.36 | ||||||||
A 5 | 0.7012 | 170.4 | A 6 | 0.6967 | 170.38 | A 7 | 1.6148 | 170.36 | ||||||||
A 5 | 0.7004 | 180.4 | A 6 | 0.6963 | 180.38 | A 7 | 1.6167 | 180.36 | ||||||||
A 5 | 0.7006 | 190.4 | A 6 | 0.6968 | 190.38 | A 7 | 1.6147 | 190.36 | ||||||||
A 5 | 0.7009 | 200.4 | A 6 | 0.696 | 200.38 | A 7 | 1.6102 | 200.36 | ||||||||
A 5 | 0.7002 | 210.4 | A 6 | 0.6957 | 210.38 | A 7 | 1.6071 | 210.36 | ||||||||
A 5 | 0.7001 | 220.4 | A 6 | 0.6958 | 220.39 | A 7 | 1.6026 | 220.36 | ||||||||
A 5 | 0.7 | 230.4 | A 6 | 0.6955 | 230.38 | A 7 | 1.6019 | 230.36 | ||||||||
A 5 | 0.6996 | 240.4 | A 6 | 0.6952 | 240.38 | A 7 | 1.5953 | 240.36 | ||||||||
A 5 | 0.6997 | 250.4 | A 6 | 0.6946 | 250.38 | A 7 | 1.5977 | 250.36 | ||||||||
A 5 | 0.7001 | 260.4 | A 6 | 0.6949 | 260.38 | A 7 | 1.5925 | 260.36 | ||||||||
A 5 | 0.6996 | 270.4 | A 6 | 0.6939 | 270.38 | A 7 | 1.6021 | 270.36 | ||||||||
A 5 | 0.6998 | 280.4 | A 6 | 0.6948 | 280.38 | A 7 | 1.5933 | 280.36 | ||||||||
A 5 | 0.6992 | 290.4 | A 6 | 0.6949 | 290.38 | A 7 | 1.5866 | 290.36 | ||||||||
A 5 | 0.6994 | 300.4 | A 6 | 0.6945 | 300.38 | A 7 | 1.5878 | 300.36 | ||||||||
B 5 | 0.6862 | 0.42 | 5.6407822765207626E-6 | 0.7801197324348885 | B 6 | 0.6901 | 0.42 | -2.3632524697855934E-5 | 0.9717508669540801 | B 7 | 1.7027 | 0.42 | -2.1954822954822938E-4 | 0.9726877232195452 | ||
B 5 | 0.692 | 10.82 | B 6 | 0.68 | 10.8 | B 7 | 1.7185 | 10.78 | ||||||||
B 5 | 0.6923 | 20.82 | B 6 | 0.6813 | 20.8 | B 7 | 1.7113 | 20.78 | ||||||||
B 5 | 0.6914 | 30.83 | B 6 | 0.6822 | 30.8 | B 7 | 1.7123 | 30.78 | ||||||||
B 5 | 0.6904 | 40.82 | B 6 | 0.6826 | 40.8 | B 7 | 1.7103 | 40.78 | ||||||||
B 5 | 0.6897 | 50.82 | B 6 | 0.6825 | 50.8 | B 7 | 1.7117 | 50.78 | ||||||||
B 5 | 0.6894 | 60.83 | B 6 | 0.6822 | 60.8 | B 7 | 1.7086 | 60.78 | ||||||||
B 5 | 0.6892 | 70.82 | B 6 | 0.6824 | 70.8 | B 7 | 1.7075 | 70.78 | ||||||||
B 5 | 0.6892 | 80.82 | B 6 | 0.682 | 80.8 | B 7 | 1.708 | 80.78 | ||||||||
B 5 | 0.6893 | 90.83 | B 6 | 0.6817 | 90.81 | B 7 | 1.7032 | 90.78 | ||||||||
B 5 | 0.6896 | 100.82 | B 6 | 0.6815 | 100.8 | B 7 | 1.701 | 100.78 | ||||||||
B 5 | 0.6893 | 110.82 | B 6 | 0.6815 | 110.8 | B 7 | 1.6958 | 110.78 | ||||||||
B 5 | 0.6891 | 120.83 | B 6 | 0.6811 | 120.8 | B 7 | 1.6883 | 120.78 | ||||||||
B 5 | 0.6897 | 130.83 | B 6 | 0.681 | 130.8 | B 7 | 1.691 | 130.78 | ||||||||
B 5 | 0.69 | 140.83 | B 6 | 0.6807 | 140.8 | B 7 | 1.6882 | 140.78 | ||||||||
B 5 | 0.6898 | 150.83 | B 6 | 0.6802 | 150.8 | B 7 | 1.6853 | 150.78 | ||||||||
B 5 | 0.6898 | 160.83 | B 6 | 0.6798 | 160.8 | B 7 | 1.6841 | 160.78 | ||||||||
B 5 | 0.6903 | 170.83 | B 6 | 0.6794 | 170.8 | B 7 | 1.6827 | 170.78 | ||||||||
B 5 | 0.6899 | 180.83 | B 6 | 0.6792 | 180.8 | B 7 | 1.6826 | 180.78 | ||||||||
B 5 | 0.6897 | 190.83 | B 6 | 0.6789 | 190.8 | B 7 | 1.6811 | 190.78 | ||||||||
B 5 | 0.6899 | 200.83 | B 6 | 0.6785 | 200.8 | B 7 | 1.676 | 200.78 | ||||||||
B 5 | 0.69 | 210.83 | B 6 | 0.6781 | 210.8 | B 7 | 1.67 | 210.78 | ||||||||
B 5 | 0.69 | 220.83 | B 6 | 0.6784 | 220.81 | B 7 | 1.6702 | 220.78 | ||||||||
B 5 | 0.6901 | 230.83 | B 6 | 0.6777 | 230.8 | B 7 | 1.6732 | 230.78 | ||||||||
B 5 | 0.69 | 240.83 | B 6 | 0.6784 | 240.8 | B 7 | 1.6705 | 240.78 | ||||||||
B 5 | 0.6905 | 250.83 | B 6 | 0.6776 | 250.8 | B 7 | 1.6653 | 250.78 | ||||||||
B 5 | 0.6907 | 260.83 | B 6 | 0.6776 | 260.8 | B 7 | 1.659 | 260.78 | ||||||||
B 5 | 0.6906 | 270.83 | B 6 | 0.6778 | 270.8 | B 7 | 1.6662 | 270.78 | ||||||||
B 5 | 0.6908 | 280.83 | B 6 | 0.6775 | 280.8 | B 7 | 1.658 | 280.78 | ||||||||
B 5 | 0.6913 | 290.83 | B 6 | 0.6771 | 290.8 | B 7 | 1.6564 | 290.78 | ||||||||
B 5 | 0.691 | 300.83 | B 6 | 0.677 | 300.8 | B 7 | 1.6604 | 300.78 | ||||||||
C 5 | 0.6997 | 0.85 | 5.350242872480166E-5 | 0.9834772466989931 | C 6 | 0.6902 | 0.85 | 2.279618350838691E-5 | 0.9582977627409738 | C 7 | 1.7291 | 0.85 | -1.9185592185592146E-4 | 0.9940423055765812 | ||
C 5 | 0.6862 | 11.24 | C 6 | 0.6919 | 11.23 | C 7 | 1.7282 | 11.21 |
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B 1 | 0.7128 | 300.86 | B 2 | 0.7187 | 300.87 | B 3 | 1.6537 | 300.86 | ||||||||
C 1 | 0.7146 | 0.89 | -2.5660527592494576E-5 | 0.8731471684635295 | C 2 | 0.7473 | 0.85 | -5.947496947496952E-5 | 0.9506572176383411 | C 3 | 1.6988 | 0.85 | -2.4297924297924305E-4 | 0.9873468529274182 | ||
C 1 | 0.7058 | 11.36 | C 2 | 0.7404 | 11.29 | C 3 | 1.6493 | 11.28 | ||||||||
C 1 | 0.7028 | 21.32 | C 2 | 0.7385 | 21.29 | C 3 | 1.6365 | 21.28 | ||||||||
C 1 | 0.6943 | 31.29 | C 2 | 0.7354 | 31.29 | C 3 | 1.6363 | 31.28 | ||||||||
C 1 | 0.6922 | 41.36 | C 2 | 0.7325 | 41.29 | C 3 | 1.6319 | 41.28 | ||||||||
C 1 | 0.6912 | 51.32 | C 2 | 0.7313 | 51.29 | C 3 | 1.6262 | 51.28 | ||||||||
C 1 | 0.6907 | 61.29 | C 2 | 0.73 | 61.29 | C 3 | 1.623 | 61.28 | ||||||||
C 1 | 0.6902 | 71.36 | C 2 | 0.7288 | 71.29 | C 3 | 1.6191 | 71.28 | ||||||||
C 1 | 0.689 | 81.32 | C 2 | 0.7285 | 81.29 | C 3 | 1.6147 | 81.28 | ||||||||
C 1 | 0.6888 | 91.29 | C 2 | 0.7271 | 91.29 | C 3 | 1.6098 | 91.28 | ||||||||
C 1 | 0.6878 | 101.36 | C 2 | 0.7268 | 101.29 | C 3 | 1.6108 | 101.28 | ||||||||
C 1 | 0.6876 | 111.32 | C 2 | 0.7259 | 111.29 | C 3 | 1.608 | 111.28 | ||||||||
C 1 | 0.6877 | 121.29 | C 2 | 0.7245 | 121.29 | C 3 | 1.6044 | 121.28 | ||||||||
C 1 | 0.6869 | 131.36 | C 2 | 0.724 | 131.29 | C 3 | 1.6001 | 131.28 | ||||||||
C 1 | 0.687 | 141.32 | C 2 | 0.7229 | 141.29 | C 3 | 1.5983 | 141.28 | ||||||||
C 1 | 0.6868 | 151.29 | C 2 | 0.7227 | 151.29 | C 3 | 1.5979 | 151.28 | ||||||||
C 1 | 0.6866 | 161.36 | C 2 | 0.7228 | 161.29 | C 3 | 1.5948 | 161.28 | ||||||||
C 1 | 0.6866 | 171.32 | C 2 | 0.7206 | 171.29 | C 3 | 1.5906 | 171.28 | ||||||||
C 1 | 0.6862 | 181.29 | C 2 | 0.7204 | 181.29 | C 3 | 1.5904 | 181.28 | ||||||||
C 1 | 0.6865 | 191.36 | C 2 | 0.7199 | 191.29 | C 3 | 1.5893 | 191.28 | ||||||||
C 1 | 0.6856 | 201.32 | C 2 | 0.7198 | 201.29 | C 3 | 1.587 | 201.28 | ||||||||
C 1 | 0.686 | 211.29 | C 2 | 0.7195 | 211.29 | C 3 | 1.5831 | 211.28 | ||||||||
C 1 | 0.6849 | 221.36 | C 2 | 0.7191 | 221.29 | C 3 | 1.5821 | 221.28 | ||||||||
C 1 | 0.685 | 231.32 | C 2 | 0.7188 | 231.29 | C 3 | 1.5817 | 231.28 | ||||||||
C 1 | 0.6847 | 241.29 | C 2 | 0.7187 | 241.29 | C 3 | 1.5773 | 241.28 | ||||||||
C 1 | 0.6851 | 251.36 | C 2 | 0.718 | 251.29 | C 3 | 1.5757 | 251.28 | ||||||||
C 1 | 0.6852 | 261.32 | C 2 | 0.7175 | 261.29 | C 3 | 1.5724 | 261.28 | ||||||||
C 1 | 0.685 | 271.29 | C 2 | 0.7175 | 271.29 | C 3 | 1.5705 | 271.28 | ||||||||
C 1 | 0.6842 | 281.36 | C 2 | 0.7171 | 281.29 | C 3 | 1.5682 | 281.28 | ||||||||
C 1 | 0.6847 | 291.32 | C 2 | 0.7168 | 291.29 | C 3 | 1.5661 | 291.28 | ||||||||
C 1 | 0.6849 | 301.29 | C 2 | 0.7162 | 301.29 | C 3 | 1.5635 | 301.28 | ||||||||
D 1 | 0.7259 | 1.35 | -4.191745485015371E-5 | 0.8165368291223859 | D 2 | 0.7433 | 1.27 | -5.806471306471307E-5 | 0.9624556476281505 | D 3 | 1.5581 | 1.28 | -3.0100732600732584E-4 | 0.997011855858767 | ||
D 1 | 0.7192 | 11.82 | D 2 | 0.7339 | 11.72 | D 3 | 1.5577 | 11.71 | ||||||||
D 1 | 0.7152 | 21.79 | D 2 | 0.7272 | 21.72 | D 3 | 1.5475 | 21.71 | ||||||||
D 1 | 0.7102 | 31.75 | D 2 | 0.7253 | 31.72 | D 3 | 1.5492 | 31.71 | ||||||||
D 1 | 0.7087 | 41.82 | D 2 | 0.7221 | 41.72 | D 3 | 1.5545 | 41.71 | ||||||||
D 1 | 0.704 | 51.78 | D 2 | 0.721 | 51.72 | D 3 | 1.5511 | 51.71 | ||||||||
D 1 | 0.7018 | 61.75 | D 2 | 0.7191 | 61.72 | D 3 | 1.5499 | 61.71 | ||||||||
D 1 | 0.7011 | 71.82 | D 2 | 0.7189 | 71.72 | D 3 | 1.5477 | 71.71 | ||||||||
D 1 | 0.7008 | 81.78 | D 2 | 0.7181 | 81.72 | D 3 | 1.5431 | 81.71 | ||||||||
D 1 | 0.6988 | 91.75 | D 2 | 0.7177 | 91.72 | D 3 | 1.5409 | 91.71 | ||||||||
D 1 | 0.6984 | 101.82 | D 2 | 0.7167 | 101.72 | D 3 | 1.5372 | 101.71 | ||||||||
D 1 | 0.6984 | 111.78 | D 2 | 0.7163 | 111.72 | D 3 | 1.5367 | 111.71 | ||||||||
D 1 | 0.6978 | 121.75 | D 2 | 0.7154 | 121.72 | D 3 | 1.5323 | 121.71 | ||||||||
D 1 | 0.6969 | 131.82 | D 2 | 0.7145 | 131.72 | D 3 | 1.5279 | 131.71 | ||||||||
D 1 | 0.6974 | 141.78 | D 2 | 0.7138 | 141.72 | D 3 | 1.5222 | 141.71 | ||||||||
D 1 | 0.6977 | 151.75 | D 2 | 0.7132 | 151.72 | D 3 | 1.5229 | 151.71 | ||||||||
D 1 | 0.6967 | 161.82 | D 2 | 0.712 | 161.72 | D 3 | 1.5197 | 161.71 | ||||||||
D 1 | 0.6965 | 171.78 | D 2 | 0.7111 | 171.72 | D 3 | 1.5158 | 171.71 | ||||||||
D 1 | 0.6965 | 181.75 | D 2 | 0.7112 | 181.72 | D 3 | 1.5139 | 181.71 | ||||||||
D 1 | 0.6959 | 191.82 | D 2 | 0.7101 | 191.72 | D 3 | 1.5107 | 191.71 | ||||||||
D 1 | 0.6956 | 201.79 | D 2 | 0.7101 | 201.72 | D 3 | 1.5052 | 201.71 | ||||||||
D 1 | 0.6951 | 211.75 | D 2 | 0.7099 | 211.72 | D 3 | 1.5041 | 211.71 | ||||||||
D 1 | 0.6947 | 221.82 | D 2 | 0.7093 | 221.72 | D 3 | 1.5029 | 221.71 | ||||||||
D 1 | 0.695 | 231.78 | D 2 | 0.7089 | 231.72 | D 3 | 1.4982 | 231.71 | ||||||||
D 1 | 0.6934 | 241.75 | D 2 | 0.7085 | 241.72 | D 3 | 1.4965 | 241.71 | ||||||||
D 1 | 0.6934 | 251.82 | D 2 | 0.7076 | 251.72 | D 3 | 1.4932 | 251.71 | ||||||||
D 1 | 0.6934 | 261.78 | D 2 | 0.7077 | 261.72 | D 3 | 1.4909 | 261.71 | ||||||||
D 1 | 0.6936 | 271.75 | D 2 | 0.708 | 271.72 | D 3 | 1.4871 | 271.71 | ||||||||
D 1 | 0.6931 | 281.82 | D 2 | 0.7071 | 281.72 | D 3 | 1.4834 | 281.71 | ||||||||
D 1 | 0.6939 | 291.79 | D 2 | 0.707 | 291.72 | D 3 | 1.4774 | 291.71 | ||||||||
D 1 | 0.6935 | 301.75 | D 2 | 0.7068 | 301.72 | D 3 | 1.4762 | 301.71 | ||||||||
E 1 | 0.7095 | 1.77 | -9.878318610699929E-6 | 0.6692791431495088 | E 2 | 0.7424 | 1.69 | -4.682539682539677E-5 | 0.9238139151788849 | E 3 | 1.6621 | 1.7 | -2.589926739926741E-4 | 0.9816912447295705 | ||
E 1 | 0.7026 | 12.24 | E 2 | 0.7435 | 12.14 | E 3 | 1.6433 | 12.13 | ||||||||
E 1 | 0.7 | 22.21 | E 2 | 0.7433 | 22.14 | E 3 | 1.6493 | 22.13 | ||||||||
E 1 | 0.6967 | 32.18 | E 2 | 0.7409 | 32.14 | E 3 | 1.6506 | 32.13 | ||||||||
E 1 | 0.6931 | 42.24 | E 2 | 0.7391 | 42.14 | E 3 | 1.646 | 42.13 | ||||||||
E 1 | 0.6931 | 52.21 | E 2 | 0.7375 | 52.14 | E 3 | 1.6466 | 52.13 | ||||||||
E 1 | 0.6925 | 62.17 | E 2 | 0.7365 | 62.14 | E 3 | 1.6463 | 62.13 | ||||||||
E 1 | 0.6924 | 72.24 | E 2 | 0.7358 | 72.14 | E 3 | 1.6427 | 72.13 | ||||||||
E 1 | 0.6923 | 82.21 | E 2 | 0.7349 | 82.14 | E 3 | 1.6424 | 82.13 | ||||||||
E 1 | 0.6919 | 92.18 | E 2 | 0.7342 | 92.14 | E 3 | 1.641 | 92.13 | ||||||||
E 1 | 0.6916 | 102.24 | E 2 | 0.7336 | 102.14 | E 3 | 1.6369 | 102.13 | ||||||||
E 1 | 0.6916 | 112.21 | E 2 | 0.7328 | 112.14 | E 3 | 1.6323 | 112.13 | ||||||||
E 1 | 0.6917 | 122.17 | E 2 | 0.7322 | 122.14 | E 3 | 1.6283 | 122.13 | ||||||||
E 1 | 0.6911 | 132.24 | E 2 | 0.7317 | 132.14 | E 3 | 1.6274 | 132.13 | ||||||||
E 1 | 0.6923 | 142.21 | E 2 | 0.731 | 142.14 | E 3 | 1.6252 | 142.13 | ||||||||
E 1 | 0.6914 | 152.17 | E 2 | 0.7308 | 152.14 | E 3 | 1.6241 | 152.13 | ||||||||
E 1 | 0.6915 | 162.24 | E 2 | 0.7301 | 162.14 | E 3 | 1.6226 | 162.13 | ||||||||
E 1 | 0.692 | 172.21 | E 2 | 0.7288 | 172.14 | E 3 | 1.6207 | 172.13 | ||||||||
E 1 | 0.6916 | 182.17 | E 2 | 0.7282 | 182.14 | E 3 | 1.6187 | 182.13 | ||||||||
E 1 | 0.6912 | 192.24 | E 2 | 0.7284 | 192.14 | E 3 | 1.6185 | 192.13 | ||||||||
E 1 | 0.6912 | 202.21 | E 2 | 0.7282 | 202.14 | E 3 | 1.6108 | 202.13 | ||||||||
E 1 | 0.6911 | 212.18 | E 2 | 0.7277 | 212.14 | E 3 | 1.6117 | 212.13 | ||||||||
E 1 | 0.6908 | 222.24 | E 2 | 0.728 | 222.14 | E 3 | 1.6082 | 222.13 | ||||||||
E 1 | 0.6914 | 232.21 | E 2 | 0.7276 | 232.14 | E 3 | 1.6057 | 232.13 | ||||||||
E 1 | 0.6891 | 242.18 | E 2 | 0.7271 | 242.14 | E 3 | 1.6001 | 242.13 | ||||||||
E 1 | 0.6896 | 252.24 | E 2 | 0.7276 | 252.14 | E 3 | 1.5948 | 252.13 | ||||||||
E 1 | 0.6902 | 262.21 | E 2 | 0.7264 | 262.14 | E 3 | 1.5913 | 262.13 | ||||||||
E 1 | 0.6903 | 272.17 | E 2 | 0.7265 | 272.14 | E 3 | 1.588 | 272.13 | ||||||||
E 1 | 0.6906 | 282.24 | E 2 | 0.7269 | 282.14 | E 3 | 1.586 | 282.13 | ||||||||
E 1 | 0.6904 | 292.21 | E 2 | 0.7254 | 292.14 | E 3 | 1.584 | 292.13 | ||||||||
E 1 | 0.6913 | 302.18 | E 2 | 0.7264 | 302.14 | E 3 | 1.5835 | 302.13 | ||||||||
F 1 | 0.7092 | 2.24 | -3.335274015524088E-5 | 0.9216262120495528 | F 2 | 0.7312 | 2.12 | -5.271062271062269E-5 | 0.9543384715615864 | |||||||
F 1 | 0.7035 | 12.71 | F 2 | 0.7238 | 12.57 | |||||||||||
F 1 | 0.6991 | 22.67 | F 2 | 0.7208 | 22.57 | |||||||||||
F 1 | 0.6958 | 32.64 | F 2 | 0.7184 | 32.57 | |||||||||||
F 1 | 0.6949 | 42.71 | F 2 | 0.7179 | 42.57 | |||||||||||
F 1 | 0.6954 | 52.67 | F 2 | 0.7168 | 52.57 |
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Kinetic assay_single mutant T249A _ pH= 8.0_ R-PEA, R-PPA, R-PBA | 9,15 Αbs/mM acetophenone | ||||||||||||||||
6,36 Abs/mM butyrophenone | |||||||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | mM/min | ||||||||||||||||
R-PEA | *60 | (ΔΑbs/min) / (9,15 Αbs/mM) | *DF | 40 | *αραίωση πηγαδιού | μmol/(ml * min) | (Bradford) | μmol/ (mg * min) | |||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ΔΑbs/min | mmol/ (L * min) | DF | mmol/ (L * min) | αραιωση πηγαδιού | mmol/ (L * min) | U /ml | STDEV | (mg/ml) | U/mg | STDEV | |
A 4 | blank (R)-PEA | -4.2631149702009156E-5 | 0.14655326822944895 | -3.53897491438189E-5 | |||||||||||||
B 4 | blank (R)-PEA | -3.53897491438189E-5 | 0.9661639780360295 | ||||||||||||||
C 4 | T249 (1:5) | 4.8545593185060875E-5 | 0.9674378973575171 | 4.8545593185060875E-5 | 0.0029127355911036526 | 3.1868004279033394E-4 | 5 | 0.0015934002139516698 | 40 | 0.0637360085580668 | 0.08166040172631177 | 0.021320818256139077 | 0.5926122774382141 | 0.13779735053637274 | 0.03597768569410375 | ||
D 4 | T249 (1:5) | 8.015639188294296E-5 | 0.9801057488543711 | 8.015639188294296E-5 | 0.004809383512976577 | 5.261907563431703E-4 | 5 | 0.002630953781715852 | 40 | 0.10523815126863408 | |||||||
E 4 | T249 (1:5) | 5.789203287661854E-5 | 0.9363339226337993 | 5.789203287661854E-5 | 0.0034735219725971125 | 3.8003522676117204E-4 | 5 | 0.0019001761338058602 | 40 | 0.0760070453522344 | |||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | |||||||||||||||||
R-PPA | |||||||||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ||||||||||||
A 5 | blank (R)-PPA | -5.8009390631792436E-5 | 0.98875420154175 | -6.54238921846089E-5 | |||||||||||||
B 5 | blank (R)-PPA | -7.283839373742536E-5 | 0.9600148882687405 | ||||||||||||||
C 5 | T249 (1:5) | -3.9614834026526895E-5 | 0.8711325391441321 | -3.9614834026526895E-5 | |||||||||||||
D 5 | T249 (1:5) | -5.2087850728540136E-5 | 0.9853664234864297 | -5.2087850728540136E-5 | |||||||||||||
E 5 | T249 (1:5) | -4.144050763478073E-5 | 0.9503536748441558 | -4.144050763478073E-5 | |||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | |||||||||||||||||
R- PBA | |||||||||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ||||||||||||
A 6 | blank (R)-PBA | -2.3820077486876035E-4 | 0.9916000804852558 | -2.2786478000985303E-4 | |||||||||||||
B 6 | blank (R)-PBA | -2.1752878515094573E-4 | 0.99399143882791 | ||||||||||||||
C 6 | T249 (1:5) | -2.2682031837384443E-4 | 0.985982286686604 | -2.2682031837384443E-4 | |||||||||||||
D 6 | T249 (1:5) | -2.1552064824847725E-4 | 0.9907648946047265 | -2.1552064824847725E-4 | |||||||||||||
E 6 | T249 (1:5) | -1.9658801870386228E-4 | 0.9036719507885937 | -1.9658801870386228E-4 | |||||||||||||
Measurements | |||||||||||||||||
T249_1;5_PEA | T249_1;5_PPA | T29_1;5_PBA | |||||||||||||||
Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | |||
A 4 | 0.7606 | 0 | -4.2631149702009156E-5 | 0.14655326822944895 | A 5 | 0.7383 | 0 | -5.8009390631792436E-5 | 0.98875420154175 | A 6 | 1.5343 | 0 | -2.3820077486876035E-4 | 0.9916000804852558 | |||
A 4 | 0.7667 | 10.41 | A 5 | 0.7254 | 10.39 | A 6 | 1.5509 | 10.37 | |||||||||
A 4 | 0.7638 | 20.41 | A 5 | 0.7261 | 20.4 | A 6 | 1.5568 | 20.37 | |||||||||
A 4 | 0.7571 | 30.41 | A 5 | 0.7251 | 30.39 | A 6 | 1.5468 | 30.37 | |||||||||
A 4 | 0.7512 | 40.41 | A 5 | 0.7246 | 40.39 | A 6 | 1.5402 | 40.38 | |||||||||
A 4 | 0.7418 | 50.41 | A 5 | 0.7246 | 50.4 | A 6 | 1.5341 | 50.37 | |||||||||
A 4 | 0.7398 | 60.42 | A 5 | 0.7237 | 60.39 | A 6 | 1.5339 | 60.37 | |||||||||
A 4 | 0.7428 | 70.41 | A 5 | 0.7224 | 70.39 | A 6 | 1.5331 | 70.38 | |||||||||
A 4 | 0.7434 | 80.42 | A 5 | 0.7223 | 80.4 | A 6 | 1.5331 | 80.37 | |||||||||
A 4 | 0.7442 | 90.41 | A 5 | 0.7217 | 90.39 | A 6 | 1.5307 | 90.37 | |||||||||
A 4 | 0.7466 | 100.41 | A 5 | 0.7216 | 100.39 | A 6 | 1.5292 | 100.37 | |||||||||
A 4 | 0.7579 | 110.41 | A 5 | 0.7206 | 110.4 | A 6 | 1.5274 | 110.37 | |||||||||
A 4 | 0.7613 | 120.42 | A 5 | 0.7195 | 120.39 | A 6 | 1.524 | 120.37 | |||||||||
A 4 | 0.7591 | 130.41 | A 5 | 0.7192 | 130.39 | A 6 | 1.5218 | 130.37 | |||||||||
A 4 | 0.7582 | 140.42 | A 5 | 0.7182 | 140.4 | A 6 | 1.5189 | 140.37 | |||||||||
A 4 | 0.7497 | 150.41 | A 5 | 0.7176 | 150.39 | A 6 | 1.5166 | 150.37 | |||||||||
A 4 | 0.7422 | 160.41 | A 5 | 0.7172 | 160.4 | A 6 | 1.5139 | 160.37 | |||||||||
A 4 | 0.7357 | 170.42 | A 5 | 0.7164 | 170.39 | A 6 | 1.512 | 170.37 | |||||||||
A 4 | 0.7355 | 180.42 | A 5 | 0.7156 | 180.39 | A 6 | 1.5091 | 180.37 | |||||||||
A 4 | 0.738 | 190.42 | A 5 | 0.7156 | 190.4 | A 6 | 1.5048 | 190.37 | |||||||||
A 4 | 0.7386 | 200.42 | A 5 | 0.7143 | 200.39 | A 6 | 1.5021 | 200.38 | |||||||||
A 4 | 0.7403 | 210.42 | A 5 | 0.7139 | 210.39 | A 6 | 1.4992 | 210.37 | |||||||||
A 4 | 0.7452 | 220.42 | A 5 | 0.7133 | 220.4 | A 6 | 1.4959 | 220.37 | |||||||||
A 4 | 0.7555 | 230.42 | A 5 | 0.713 | 230.39 | A 6 | 1.495 | 230.37 | |||||||||
A 4 | 0.7538 | 240.42 | A 5 | 0.7127 | 240.39 | A 6 | 1.4929 | 240.37 | |||||||||
A 4 | 0.75 | 250.41 | A 5 | 0.7124 | 250.4 | A 6 | 1.4907 | 250.37 | |||||||||
A 4 | 0.7428 | 260.41 | A 5 | 0.712 | 260.39 | A 6 | 1.4898 | 260.38 | |||||||||
A 4 | 0.7358 | 270.42 | A 5 | 0.7113 | 270.39 | A 6 | 1.4866 | 270.37 | |||||||||
A 4 | 0.732 | 280.42 | A 5 | 0.711 | 280.4 | A 6 | 1.4843 | 280.37 | |||||||||
A 4 | 0.7319 | 290.42 | A 5 | 0.7104 | 290.39 | A 6 | 1.4818 | 290.37 | |||||||||
A 4 | 0.7315 | 300.42 | A 5 | 0.7104 | 300.4 | A 6 | 1.479 | 300.37 | |||||||||
B 4 | 0.7396 | 0.43 | -3.53897491438189E-5 | 0.9661639780360295 | B 5 | 0.7569 | 0.42 | -7.283839373742536E-5 | 0.9600148882687405 | B 6 | 1.5557 | 0.42 | -2.1752878515094573E-4 | 0.99399143882791 | |||
B 4 | 0.743 | 10.84 | B 5 | 0.747 | 10.82 | B 6 | 1.549 | 10.79 | |||||||||
B 4 | 0.7423 | 20.84 | B 5 | 0.746 | 20.83 | B 6 | 1.5526 | 20.79 | |||||||||
B 4 | 0.7415 | 30.84 | B 5 | 0.7445 | 30.82 | B 6 | 1.5478 | 30.79 | |||||||||
B 4 | 0.7402 | 40.84 | B 5 | 0.743 | 40.82 | B 6 | 1.5416 | 40.8 | |||||||||
B 4 | 0.7387 | 50.84 | B 5 | 0.7416 | 50.83 | B 6 | 1.5352 | 50.79 | |||||||||
B 4 | 0.7385 | 60.85 | B 5 | 0.7404 | 60.82 | B 6 | 1.5331 | 60.79 | |||||||||
B 4 | 0.7377 | 70.84 | B 5 | 0.7389 | 70.82 | B 6 | 1.5307 | 70.8 | |||||||||
B 4 | 0.7372 | 80.85 | B 5 | 0.7383 | 80.83 | B 6 | 1.5287 | 80.79 | |||||||||
B 4 | 0.7368 | 90.84 | B 5 | 0.737 | 90.82 | B 6 | 1.528 | 90.79 | |||||||||
B 4 | 0.7364 | 100.84 | B 5 | 0.7363 | 100.82 | B 6 | 1.5244 | 100.79 | |||||||||
B 4 | 0.7357 | 110.84 | B 5 | 0.7348 | 110.83 | B 6 | 1.5219 | 110.79 | |||||||||
B 4 | 0.7357 | 120.85 | B 5 | 0.734 | 120.82 | B 6 | 1.5199 | 120.79 | |||||||||
B 4 | 0.7353 | 130.84 | B 5 | 0.733 | 130.82 | B 6 | 1.5176 | 130.79 | |||||||||
B 4 | 0.7348 | 140.85 | B 5 | 0.7322 | 140.83 | B 6 | 1.5157 | 140.79 | |||||||||
B 4 | 0.7341 | 150.84 | B 5 | 0.7311 | 150.82 | B 6 | 1.514 | 150.79 | |||||||||
B 4 | 0.7337 | 160.84 | B 5 | 0.7307 | 160.83 | B 6 | 1.5116 | 160.79 | |||||||||
B 4 | 0.7334 | 170.85 | B 5 | 0.7299 | 170.82 | B 6 | 1.5092 | 170.79 | |||||||||
B 4 | 0.7331 | 180.85 | B 5 | 0.7288 | 180.82 | B 6 | 1.5066 | 180.79 | |||||||||
B 4 | 0.7327 | 190.85 | B 5 | 0.7286 | 190.83 | B 6 | 1.5034 | 190.79 | |||||||||
B 4 | 0.7323 | 200.85 | B 5 | 0.7275 | 200.82 | B 6 | 1.5007 | 200.8 | |||||||||
B 4 | 0.7325 | 210.85 | B 5 | 0.7268 | 210.82 | B 6 | 1.499 | 210.79 | |||||||||
B 4 | 0.7322 | 220.85 | B 5 | 0.7271 | 220.83 | B 6 | 1.4965 | 220.79 | |||||||||
B 4 | 0.7319 | 230.85 | B 5 | 0.7267 | 230.82 | B 6 | 1.4949 | 230.79 | |||||||||
B 4 | 0.7318 | 240.85 | B 5 | 0.7259 | 240.82 | B 6 | 1.493 | 240.79 | |||||||||
B 4 | 0.7315 | 250.84 | B 5 | 0.7257 | 250.83 | B 6 | 1.4931 | 250.79 | |||||||||
B 4 | 0.731 | 260.84 | B 5 | 0.7253 | 260.82 | B 6 | 1.4908 | 260.8 | |||||||||
B 4 | 0.7308 | 270.85 | B 5 | 0.7247 | 270.82 | B 6 | 1.4888 | 270.79 | |||||||||
B 4 | 0.7308 | 280.85 | B 5 | 0.7243 | 280.83 | B 6 | 1.4872 | 280.79 | |||||||||
B 4 | 0.7301 | 290.85 | B 5 | 0.7242 | 290.82 | B 6 | 1.4849 | 290.79 | |||||||||
B 4 | 0.7302 | 300.85 | B 5 | 0.7233 | 300.83 | B 6 | 1.4823 | 300.79 | |||||||||
C 4 | 0.7316 | 0.85 | 4.8545593185060875E-5 | 0.9674378973575171 | C 5 | 0.7334 | 0.85 | -3.9614834026526895E-5 | 0.8711325391441321 | C 6 | 1.5141 | 0.85 | -2.2682031837384443E-4 | 0.985982286686604 | |||
C 4 | 0.7317 | 11.26 | C 5 | 0.725 | 11.24 | C 6 | 1.5103 | 11.22 | |||||||||
C 4 | 0.7273 | 21.26 | C 5 | 0.7233 | 21.25 | C 6 | 1.5105 | 21.22 | |||||||||
C 4 | 0.7259 | 31.26 | C 5 | 0.7272 | 31.24 | C 6 | 1.5097 | 31.22 | |||||||||
C 4 | 0.7264 | 41.26 | C 5 | 0.722 | 41.24 | C 6 | 1.5048 | 41.23 |
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Kinetic assay_duble mutants V37A/S248A , V37A/T249A pH= 8.0_ R-PEA, R-PPA, R-PBA | |||||||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | |||||||||||||||||
R- phenylethylamine | |||||||||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ||||||||||||
A 9 | blank | -2.8152683550859125E-6 | 0.26242700643215355 | ||||||||||||||
B 9 | blank | 1.4554147852293888E-7 | 1.9590323375686086E-4 | ||||||||||||||
C 9 | V37A/S248A | -8.99993172040422E-6 | 0.2255724467951563 | ||||||||||||||
D 9 | V37A/S248A | -1.031489516941991E-5 | 0.891813690223811 | ||||||||||||||
E 9 | V37A/S248A | -2.1162413891691294E-5 | 0.8877010401021397 | ||||||||||||||
F 9 | V37A/T249A | 2.0297333129148845E-6 | 0.10030075404718884 | ||||||||||||||
G 9 | V37A/T249A | -1.7237657198580164E-6 | 0.047940685227337534 | ||||||||||||||
H 9 | V37A/T249A | 3.95392726241263E-6 | 0.38505480647822266 | ||||||||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | mM/min | ||||||||||||||||
R- phenylpropylamine | *60 | (ΔΑbs/min) / (9,15 Αbs/mM) | 10 | *αραίωση πηγαδιού | μmol/(ml * min) | (Bradford) | μmol/ (mg * min) | ||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ΔΑbs/min | mmol/ (L * min) | αραιωση πηγαδιού | mmol/ (L * min) | U /ml | STDEV | (mg/ml) | U/mg | STDEV | |||
A 5 | blank | -2.1046153846153798E-5 | 0.7384085082003528 | 0 | |||||||||||||
B 5 | blank | -5.661538461538513E-6 | 0.13807532233956418 | ||||||||||||||
C 5 | V37A/S248A | -2.3092307692307673E-5 | 0.9084359103386661 | ||||||||||||||
D 5 | V37A/S248A | -2.329241371224123E-5 | 0.7991011186210762 | ||||||||||||||
E 5 | V37A/S248A | -2.0430769230769315E-5 | 0.8847372904398769 | ||||||||||||||
F 5 | V37A/T249A | 8.653846153846097E-6 | 0.7676931083677873 | 8.653846153846097E-6 | 5.192307692307659E-4 | 8.164005805515186E-5 | 10 | 8.164005805515185E-4 | 0.0011504596032897907 | 4.6548115461454085E-4 | 0.64171331857831 | 0.0017927937133026747 | 7.25372438343333E-4 | ||||
G 5 | V37A/T249A | 1.7830769230769164E-5 | 0.9527477796329081 | 1.7830769230769164E-5 | 0.0010698461538461497 | 1.6821480406386E-4 | 10 | 0.0016821480406386002 | |||||||||
H 5 | V37A/T249A | 1.0100000000000083E-5 | 0.41177000273244935 | 1.0100000000000083E-5 | 6.06000000000005E-4 | 9.52830188679253E-5 | 10 | 9.52830188679253E-4 | |||||||||
HEPES 50 mM pH 8.0 0,05 mM PLP | |||||||||||||||||
R- phenylbutylamine | |||||||||||||||||
Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | ||||||||||||
A 7 | blank | ||||||||||||||||
B 7 | blank | ||||||||||||||||
C 7 | V37A/S248A | ||||||||||||||||
D 7 | V37A/S248A | ||||||||||||||||
E 7 | V37A/S248A | ||||||||||||||||
F 7 | V37A/T249A | ||||||||||||||||
G 7 | V37A/T249A | ||||||||||||||||
H 7 | V37A/T249A | ||||||||||||||||
Measurments | |||||||||||||||||
double mutants_R-PEA | double mutants_R-PPA | double mutants_R-PBA | |||||||||||||||
Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | Well | Raw absorbance | Measurement time(s) | SLOPE sample | R squared | |||
A 9 | 0.5603 | 0 | -2.8152683550859125E-6 | 0.26242700643215355 | A 5 | 0.539 | 0 | -2.1046153846153798E-5 | 0.7384085082003528 | A 7 | 1.4259 | 0 | -5.186867090181742E-4 | 0.9578180506159945 | |||
A 9 | 0.5598 | 10.31 | A 5 | 0.5323 | 10.39 | A 7 | 1.3776 | 10.37 | |||||||||
A 9 | 0.5609 | 20.29 | A 5 | 0.5313 | 20.39 | A 7 | 1.3632 | 20.3 | |||||||||
A 9 | 0.562 | 30.26 | A 5 | 0.5231 | 30.39 | A 7 | 1.3559 | 30.36 | |||||||||
A 9 | 0.5619 | 40.23 | A 5 | 0.5223 | 40.39 | A 7 | 1.3528 | 40.29 | |||||||||
A 9 | 0.562 | 50.31 | A 5 | 0.5205 | 50.39 | A 7 | 1.356 | 50.36 | |||||||||
A 9 | 0.5629 | 60.29 | A 5 | 0.5187 | 60.39 | A 7 | 1.3486 | 60.29 | |||||||||
A 9 | 0.563 | 70.26 | A 5 | 0.5159 | 70.39 | A 7 | 1.3449 | 70.36 | |||||||||
A 9 | 0.5634 | 80.23 | A 5 | 0.5151 | 80.39 | A 7 | 1.3375 | 80.29 | |||||||||
A 9 | 0.5633 | 90.31 | A 5 | 0.5158 | 90.39 | A 7 | 1.3359 | 90.36 | |||||||||
A 9 | 0.5638 | 100.29 | A 5 | 0.5148 | 100.39 | A 7 | 1.3288 | 100.29 | |||||||||
A 9 | 0.5632 | 110.26 | A 5 | 0.5142 | 110.39 | A 7 | 1.3261 | 110.36 | |||||||||
A 9 | 0.5626 | 120.23 | A 5 | 0.5134 | 120.39 | A 7 | 1.3175 | 120.29 | |||||||||
A 9 | 0.5623 | 130.31 | A 5 | 0.5133 | 130.39 | A 7 | 1.3129 | 130.36 | |||||||||
A 9 | 0.5631 | 140.29 | A 5 | 0.5136 | 140.39 | A 7 | 1.3086 | 140.29 | |||||||||
A 9 | 0.5625 | 150.26 | A 5 | 0.5132 | 150.39 | A 7 | 1.3022 | 150.36 | |||||||||
A 9 | 0.5624 | 160.23 | A 5 | 0.5132 | 160.39 | A 7 | 1.2971 | 160.29 | |||||||||
A 9 | 0.5626 | 170.31 | A 5 | 0.5138 | 170.39 | A 7 | 1.2885 | 170.36 | |||||||||
A 9 | 0.5629 | 180.28 | A 5 | 0.5126 | 180.39 | A 7 | 1.2776 | 180.29 | |||||||||
A 9 | 0.563 | 190.26 | A 5 | 0.5116 | 190.39 | A 7 | 1.2803 | 190.36 | |||||||||
A 9 | 0.5632 | 200.23 | A 5 | 0.5118 | 200.39 | A 7 | 1.2701 | 200.29 | |||||||||
A 9 | 0.5626 | 210.31 | A 5 | 0.5118 | 210.39 | A 7 | 1.2665 | 210.36 | |||||||||
A 9 | 0.5628 | 220.29 | A 5 | 0.5117 | 220.39 | A 7 | 1.2645 | 220.29 | |||||||||
A 9 | 0.5628 | 230.26 | A 5 | 0.5116 | 230.39 | A 7 | 1.2614 | 230.37 | |||||||||
A 9 | 0.5625 | 240.23 | A 5 | 0.5116 | 240.39 | A 7 | 1.2499 | 240.3 | |||||||||
A 9 | 0.5624 | 250.31 | A 5 | 0.5116 | 250.39 | A 7 | 1.252 | 250.36 | |||||||||
A 9 | 0.5633 | 260.29 | A 5 | 0.5121 | 260.39 | A 7 | 1.2482 | 260.29 | |||||||||
A 9 | 0.5627 | 270.26 | A 5 | 0.5121 | 270.39 | A 7 | 1.2471 | 270.36 | |||||||||
A 9 | 0.5622 | 280.23 | A 5 | 0.5119 | 280.39 | A 7 | 1.2443 | 280.29 | |||||||||
A 9 | 0.5624 | 290.31 | A 5 | 0.5115 | 290.39 | A 7 | 1.2461 | 290.36 | |||||||||
A 9 | 0.5624 | 300.29 | A 5 | 0.5117 | 300.39 | A 7 | 1.2365 | 300.29 | |||||||||
B 9 | 0.5664 | 0.42 | 1.4554147852293888E-7 | 1.9590323375686086E-4 | B 5 | 0.5244 | 0.42 | -5.661538461538513E-6 | 0.13807532233956418 | B 7 | 1.4304 | 0.43 | -3.605671469805001E-4 | 0.9088649920415045 | |||
B 9 | 0.5662 | 10.74 | B 5 | 0.5199 | 10.82 | B 7 | 1.4178 | 10.79 | |||||||||
B 9 | 0.5687 | 20.71 | B 5 | 0.5188 | 20.82 | B 7 | 1.403 | 20.73 | |||||||||
B 9 | 0.5686 | 30.68 | B 5 | 0.5214 | 30.82 | B 7 | 1.3964 | 30.78 | |||||||||
B 9 | 0.5688 | 40.65 | B 5 | 0.5176 | 40.82 | B 7 | 1.3798 | 40.72 | |||||||||
B 9 | 0.5683 | 50.74 | B 5 | 0.5164 | 50.82 | B 7 | 1.3775 | 50.78 | |||||||||
B 9 | 0.5679 | 60.71 | B 5 | 0.5146 | 60.82 | B 7 | 1.371 | 60.72 | |||||||||
B 9 | 0.5679 | 70.68 | B 5 | 0.5147 | 70.82 | B 7 | 1.3651 | 70.78 | |||||||||
B 9 | 0.5675 | 80.65 | B 5 | 0.5141 | 80.82 | B 7 | 1.3586 | 80.72 | |||||||||
B 9 | 0.567 | 90.74 | B 5 | 0.5136 | 90.82 | B 7 | 1.3557 | 90.78 | |||||||||
B 9 | 0.5661 | 100.71 | B 5 | 0.5134 | 100.82 | B 7 | 1.3539 | 100.72 | |||||||||
B 9 | 0.5663 | 110.68 | B 5 | 0.513 | 110.82 | B 7 | 1.352 | 110.78 | |||||||||
B 9 | 0.5663 | 120.65 | B 5 | 0.5132 | 120.82 | B 7 | 1.3467 | 120.72 | |||||||||
B 9 | 0.5653 | 130.74 | B 5 | 0.5129 | 130.82 | B 7 | 1.3425 | 130.78 | |||||||||
B 9 | 0.5662 | 140.71 | B 5 | 0.5129 | 140.82 | B 7 | 1.3415 | 140.72 | |||||||||
B 9 | 0.5654 | 150.68 | B 5 | 0.5126 | 150.82 | B 7 | 1.3387 | 150.78 | |||||||||
B 9 | 0.5663 | 160.65 | B 5 | 0.5127 | 160.82 | B 7 | 1.3353 | 160.72 | |||||||||
B 9 | 0.5664 | 170.74 | B 5 | 0.5129 | 170.82 | B 7 | 1.3324 | 170.78 | |||||||||
B 9 | 0.5664 | 180.71 | B 5 | 0.5128 | 180.82 | B 7 | 1.3227 | 180.72 | |||||||||
B 9 | 0.5667 | 190.68 | B 5 | 0.5157 | 190.82 | B 7 | 1.3233 | 190.78 | |||||||||
B 9 | 0.5668 | 200.65 | B 5 | 0.5167 | 200.82 | B 7 | 1.3229 | 200.72 | |||||||||
B 9 | 0.5672 | 210.74 | B 5 | 0.5135 | 210.82 | B 7 | 1.3197 | 210.78 | |||||||||
B 9 | 0.567 | 220.71 | B 5 | 0.5135 | 220.82 | B 7 | 1.3177 | 220.72 | |||||||||
B 9 | 0.5673 | 230.68 | B 5 | 0.5122 | 230.82 | B 7 | 1.317 | 230.79 | |||||||||
B 9 | 0.5674 | 240.65 | B 5 | 0.5121 | 240.82 | B 7 | 1.3137 | 240.73 | |||||||||
B 9 | 0.5673 | 250.74 | B 5 | 0.5119 | 250.82 | B 7 | 1.3128 | 250.78 |