Table 1 Kinetic parameters of enzymes used in [19F]FDT synthesis

From: Distributable, metabolic PET reporting of tuberculosis

Enzyme

Substrate

KM (mM)

kcat (s-1)

kcat/KM (mM-1s-1)

OtsA a

UDP-Glc

1.7 ± 0.3

34 ± 1

20

OtsA a

G6P

7.3 ± 0.6

34 ± 1

4.7

TreT P. horikoshii b

UDP-Glc

4.6 ± 0.8

25 ± 2

5.5

TreT P. horikoshii b

Glc

8.1 ± 0.6

13.6 ± 0.4

1.7

TreT T. tenax c

UDP-Glc

2.3 ± 0.7

13.5 ± 1.4

6.2

TreT T. tenax b

Glc

5.0 ± 1.3

10.3 ± 1.3

2.1

OtsAB-fusion d

UDP-Glc

7.2 ± 5.7

-

-

OtsB e

T6P

3.6 ± 0.8

286 ± 34

79

OtsASf b

UDP-Glc

1.4 ± 0.2

63.5 ± 2.5

45.3

OtsASf b

G6P

4.9 ± 0.5

57.5 ± 2.3

11.9

OtsBSf e

T6P

2.8 ± 0.7

740 ± 96

264

  1. a Taken from reference. 46
  2. b Determined via continuous, coupled-enzyme spectrophotometric assay41 using decrease in NADH absorbance at 340 nm (ε340 = 6220 M-1cm-1) catalyzed by linked spectrophotometric assay at 25 °C.
  3. c Determined via endpoint, HPLC method with fixed Glc concentration at 70 °C.
  4. d Determined via endpoint, HPLC method with fixed Glc concentration at 37 °C.
  5. e Determined using continuous 1H NMR assay at 37 °C. See Supplementary Methods for further details. OtsASf and OtsBSf denote variants expressed from Spodoptera frugiperda rather than E. coli.