Selected Cell
Cell:
Value:
Introduction
ResultsTable
Bradford assays
LS_ATA WT
SingleMutant V37A
SingleMutant S248A
SingleMutant T249A
Double mutants
A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
Q
R
S
T
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
| 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 | ||||||||||||||||||
A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
| 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 | _ | ||||||||
A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
| 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 |
A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
Q
R
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
| 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 |
A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
Q
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
| 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 |
A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
Q
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
| Kinetic assay_single mutant S248A _ pH= 8.0_ R-PEA, R-PPA, R-PBA | ||||||||||||||||
| HEPES 50 mM pH 8.0 0,05 mM PLP | ||||||||||||||||
| R-PEA | ||||||||||||||||
| Well | Sample | ΔΑbs/s | R sqrd | ΔΑbs/s (blank) M.O | ΔΑbs/s (sample-blank) | |||||||||||
| A 1 | blank (R)-PEA | 2.166735364859991E-6 | 0.1802165137303058 | |||||||||||||
| B 1 | blank (R)-PEA | -2.2522388779618927E-5 | 0.9338607674512869 | -2.2522388779618927E-5 | ||||||||||||
| C 1 | S248 (1:5) | -2.5660527592494576E-5 | 0.8731471684635295 | -2.5660527592494576E-5 | ||||||||||||
| D 1 | S248 (1:5) | -4.191745485015371E-5 | 0.8165368291223859 | -4.191745485015371E-5 | ||||||||||||
| E 1 | S248 (1:5) | -9.878318610699929E-6 | 0.6692791431495088 | -9.878318610699929E-6 | ||||||||||||
| F 1 | S248 not diluted | -3.335274015524088E-5 | 0.9216262120495528 | -3.335274015524088E-5 | ||||||||||||
| G 1 | S248 not diluted | -2.875488811496776E-5 | 0.908009433117033 | -2.875488811496776E-5 | ||||||||||||
| H 1 | S248 not diluted | -1.2527124281758425E-5 | 0.628829639979381 | -1.2527124281758425E-5 | ||||||||||||
| 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 | -6.650183150183142E-5 | 0.9640948574251315 | -5.9432234432234424E-5 | ||||||||||||
| B 2 | blank (R)-PPA | -5.2362637362637424E-5 | 0.9770939116836415 | |||||||||||||
| C 2 | S248 (1:5) | -5.947496947496952E-5 | 0.9506572176383411 | -5.947496947496952E-5 | ||||||||||||
| D 2 | S248 (1:5) | -5.806471306471307E-5 | 0.9624556476281505 | -5.806471306471307E-5 | ||||||||||||
| E 2 | S248 (1:5) | -4.682539682539677E-5 | 0.9238139151788849 | -4.682539682539677E-5 | ||||||||||||
| F 2 | S248 not diluted | -5.271062271062269E-5 | 0.9543384715615864 | -5.271062271062269E-5 | ||||||||||||
| G 2 | S248 not diluted | -4.149572649572659E-5 | 0.9426102849112168 | -4.149572649572659E-5 | ||||||||||||
| H 2 | S248 not diluted | -3.883394383394385E-5 | 0.9572792391339422 | -3.883394383394385E-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 | -3.3330280830280826E-4 | 0.9898591650708541 | -3.51510989010989E-4 | ||||||||||||
| B 3 | blank (R)-PBA | -3.697191697191698E-4 | 0.9880400166589185 | |||||||||||||
| C 3 | GamPro S248 (1:5) | -2.4297924297924305E-4 | 0.9873468529274182 | -2.4297924297924305E-4 | ||||||||||||
| D 3 | GamPro S248 (1:5) | -3.0100732600732584E-4 | 0.997011855858767 | -3.0100732600732584E-4 | ||||||||||||
| E 3 | GamPro S248 (1:5) | -2.589926739926741E-4 | 0.9816912447295705 | -2.589926739926741E-4 | ||||||||||||
| Measurements | ||||||||||||||||
| S248_PEA | S248_PPA | S248_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 1 | 0.7085 | 0 | 2.166735364859991E-6 | 0.1802165137303058 | A 2 | 0.7352 | 0 | -6.650183150183142E-5 | 0.9640948574251315 | A 3 | 1.809 | 0 | -3.3330280830280826E-4 | 0.9898591650708541 | ||
| A 1 | 0.7106 | 10.47 | A 2 | 0.7425 | 10.44 | A 3 | 1.8058 | 10.43 | ||||||||
| A 1 | 0.71 | 20.44 | A 2 | 0.7383 | 20.44 | A 3 | 1.8028 | 20.43 | ||||||||
| A 1 | 0.7064 | 30.4 | A 2 | 0.7364 | 30.44 | A 3 | 1.8107 | 30.43 | ||||||||
| A 1 | 0.7067 | 40.47 | A 2 | 0.7359 | 40.44 | A 3 | 1.8074 | 40.43 | ||||||||
| A 1 | 0.7064 | 50.44 | A 2 | 0.7353 | 50.44 | A 3 | 1.804 | 50.43 | ||||||||
| A 1 | 0.706 | 60.4 | A 2 | 0.7334 | 60.44 | A 3 | 1.8037 | 60.43 | ||||||||
| A 1 | 0.7053 | 70.47 | A 2 | 0.7329 | 70.44 | A 3 | 1.7969 | 70.43 | ||||||||
| A 1 | 0.7046 | 80.44 | A 2 | 0.7319 | 80.44 | A 3 | 1.7961 | 80.43 | ||||||||
| A 1 | 0.7057 | 90.4 | A 2 | 0.7303 | 90.44 | A 3 | 1.7904 | 90.43 | ||||||||
| A 1 | 0.7054 | 100.47 | A 2 | 0.7296 | 100.44 | A 3 | 1.7866 | 100.43 | ||||||||
| A 1 | 0.7058 | 110.44 | A 2 | 0.7283 | 110.44 | A 3 | 1.7823 | 110.43 | ||||||||
| A 1 | 0.706 | 120.4 | A 2 | 0.7272 | 120.44 | A 3 | 1.777 | 120.43 | ||||||||
| A 1 | 0.7054 | 130.47 | A 2 | 0.7269 | 130.44 | A 3 | 1.7751 | 130.43 | ||||||||
| A 1 | 0.7061 | 140.44 | A 2 | 0.7264 | 140.44 | A 3 | 1.7727 | 140.43 | ||||||||
| A 1 | 0.7061 | 150.4 | A 2 | 0.7255 | 150.44 | A 3 | 1.7804 | 150.43 | ||||||||
| A 1 | 0.7063 | 160.47 | A 2 | 0.7246 | 160.44 | A 3 | 1.7701 | 160.43 | ||||||||
| A 1 | 0.7062 | 170.44 | A 2 | 0.7244 | 170.44 | A 3 | 1.7645 | 170.43 | ||||||||
| A 1 | 0.7062 | 180.4 | A 2 | 0.7235 | 180.44 | A 3 | 1.7613 | 180.43 | ||||||||
| A 1 | 0.7062 | 190.47 | A 2 | 0.7228 | 190.44 | A 3 | 1.7602 | 190.43 | ||||||||
| A 1 | 0.7063 | 200.44 | A 2 | 0.723 | 200.44 | A 3 | 1.7517 | 200.43 | ||||||||
| A 1 | 0.7073 | 210.4 | A 2 | 0.722 | 210.44 | A 3 | 1.7464 | 210.43 | ||||||||
| A 1 | 0.7064 | 220.47 | A 2 | 0.7217 | 220.44 | A 3 | 1.7461 | 220.43 | ||||||||
| A 1 | 0.7068 | 230.44 | A 2 | 0.7211 | 230.44 | A 3 | 1.7435 | 230.43 | ||||||||
| A 1 | 0.7078 | 240.4 | A 2 | 0.7208 | 240.44 | A 3 | 1.7407 | 240.43 | ||||||||
| A 1 | 0.7069 | 250.47 | A 2 | 0.7203 | 250.44 | A 3 | 1.736 | 250.43 | ||||||||
| A 1 | 0.7057 | 260.44 | A 2 | 0.7196 | 260.44 | A 3 | 1.7344 | 260.43 | ||||||||
| A 1 | 0.706 | 270.4 | A 2 | 0.7196 | 270.44 | A 3 | 1.7306 | 270.43 | ||||||||
| A 1 | 0.7055 | 280.47 | A 2 | 0.7185 | 280.44 | A 3 | 1.7275 | 280.43 | ||||||||
| A 1 | 0.7055 | 290.44 | A 2 | 0.7186 | 290.44 | A 3 | 1.725 | 290.43 | ||||||||
| A 1 | 0.7061 | 300.4 | A 2 | 0.718 | 300.44 | A 3 | 1.724 | 300.43 | ||||||||
| B 1 | 0.7201 | 0.46 | -2.2522388779618927E-5 | 0.9338607674512869 | B 2 | 0.7363 | 0.42 | -5.2362637362637424E-5 | 0.9770939116836415 | B 3 | 1.7503 | 0.43 | -3.697191697191698E-4 | 0.9880400166589185 | ||
| B 1 | 0.7226 | 10.93 | B 2 | 0.735 | 10.87 | B 3 | 1.7652 | 10.86 | ||||||||
| B 1 | 0.7209 | 20.9 | B 2 | 0.7348 | 20.87 | B 3 | 1.7591 | 20.86 | ||||||||
| B 1 | 0.7197 | 30.86 | B 2 | 0.734 | 30.87 | B 3 | 1.7553 | 30.86 | ||||||||
| B 1 | 0.7192 | 40.93 | B 2 | 0.7326 | 40.87 | B 3 | 1.7476 | 40.86 | ||||||||
| B 1 | 0.718 | 50.9 | B 2 | 0.7317 | 50.87 | B 3 | 1.7412 | 50.86 | ||||||||
| B 1 | 0.7172 | 60.86 | B 2 | 0.7308 | 60.87 | B 3 | 1.7344 | 60.86 | ||||||||
| B 1 | 0.7176 | 70.93 | B 2 | 0.7301 | 70.87 | B 3 | 1.7316 | 70.86 | ||||||||
| B 1 | 0.716 | 80.9 | B 2 | 0.7293 | 80.87 | B 3 | 1.7299 | 80.86 | ||||||||
| B 1 | 0.7166 | 90.86 | B 2 | 0.7286 | 90.87 | B 3 | 1.7273 | 90.86 | ||||||||
| B 1 | 0.7165 | 100.93 | B 2 | 0.7278 | 100.87 | B 3 | 1.7278 | 100.86 | ||||||||
| B 1 | 0.7164 | 110.9 | B 2 | 0.7274 | 110.87 | B 3 | 1.7272 | 110.86 | ||||||||
| B 1 | 0.7162 | 120.86 | B 2 | 0.7272 | 120.87 | B 3 | 1.7245 | 120.86 | ||||||||
| B 1 | 0.7154 | 130.93 | B 2 | 0.7261 | 130.87 | B 3 | 1.72 | 130.86 | ||||||||
| B 1 | 0.7154 | 140.9 | B 2 | 0.7253 | 140.87 | B 3 | 1.7149 | 140.86 | ||||||||
| B 1 | 0.7152 | 150.86 | B 2 | 0.725 | 150.87 | B 3 | 1.7057 | 150.86 | ||||||||
| B 1 | 0.7147 | 160.93 | B 2 | 0.7242 | 160.87 | B 3 | 1.7014 | 160.86 | ||||||||
| B 1 | 0.7146 | 170.9 | B 2 | 0.7235 | 170.87 | B 3 | 1.6988 | 170.86 | ||||||||
| B 1 | 0.7145 | 180.86 | B 2 | 0.723 | 180.87 | B 3 | 1.6942 | 180.86 | ||||||||
| B 1 | 0.7141 | 190.93 | B 2 | 0.7228 | 190.87 | B 3 | 1.6902 | 190.86 | ||||||||
| B 1 | 0.7142 | 200.9 | B 2 | 0.7225 | 200.87 | B 3 | 1.6883 | 200.86 | ||||||||
| B 1 | 0.7143 | 210.86 | B 2 | 0.7217 | 210.87 | B 3 | 1.6827 | 210.86 | ||||||||
| B 1 | 0.7138 | 220.93 | B 2 | 0.7218 | 220.87 | B 3 | 1.6781 | 220.86 | ||||||||
| B 1 | 0.7134 | 230.9 | B 2 | 0.7213 | 230.87 | B 3 | 1.6743 | 230.86 | ||||||||
| B 1 | 0.7141 | 240.86 | B 2 | 0.7209 | 240.87 | B 3 | 1.6708 | 240.86 | ||||||||
| B 1 | 0.7137 | 250.93 | B 2 | 0.7201 | 250.87 | B 3 | 1.6666 | 250.86 | ||||||||
| B 1 | 0.7128 | 260.9 | B 2 | 0.7199 | 260.87 | B 3 | 1.663 | 260.86 | ||||||||
| B 1 | 0.7125 | 270.86 | B 2 | 0.72 | 270.87 | B 3 | 1.6616 | 270.86 | ||||||||
| B 1 | 0.7119 | 280.93 | B 2 | 0.7191 | 280.87 | B 3 | 1.6596 | 280.86 | ||||||||
| B 1 | 0.7124 | 290.9 | B 2 | 0.7191 | 290.87 | B 3 | 1.6571 | 290.86 |
A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
Q
R
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
| 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 |
A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
Q
R
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
| 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 |
