Table 3 Kinetics of GOOX-VN mutant enzymes on cello- and xylo-oligosaccharides

Glucose

k_cat (min^-1)

361 ± 5

-

-

472 ± 6

(1,280 ± 70)^*

-

-

(713 ± 14)

-

K_m (mM)

15 ± 1

-

-

62 ± 2

(890 ± 60)

-

-

(325 ± 10)

-

k_cat/K_m (mM^-1 min^-1)

24 ± 2

-

-

7.7 ± 0.3

(1.4 ± 0.1)

-

-

(2.2 ± 0.1)

-

Cellobiose

k_cat (min^-1)

372 ± 14

497 ± 7

644 ± 8

480 ± 14

1,040 ± 30

117 ± 2

16 ± 1

916 ± 13

210 ± 4

K_m (mM)

0.05 ± 0.01

1.3 ± 0.1

2.5 ± 0.1

0.17 ± 0.02

13 ± 1

11.3 ± 0.4

1.1 ± 0.2

0.63 ± 0.03

8.8 ± 0.3

k_cat/K_m (mM^-1 min^-1)

7,000 ± 1,000

370 ± 20

250 ± 12

2,800 ± 300

79 ± 5

10.3 ± 0.4

15 ± 3

1,450 ± 80

24 ± 1

Cellotriose

k_cat (min^-1)

436 ± 14

533 ± 5

695 ± 6

465 ± 17

1,057 ± 15

124 ± 2

20.0 ± 0.3

1,032 ± 12

245 ± 3

K_m (mM)

0.08 ± 0.01

0.62 ± 0.02

1.17 ± 0.03

0.11 ± 0.01

6.6 ± 0.2

6.2 ± 0.2

0.57 ± 0.03

0.20 ± 0.01

4.6 ± 0.1

k_cat/K_m (mM^-1 min^-1)

5,600 ± 600

860 ± 30

596 ± 15

4,300 ± 500

160 ± 5

20 ± 1

35 ± 2

5,000 ± 200

54 ± 2

Xylose

k_cat (min^-1)

330 ± 8

-

-

189 ± 5

-

-

-

-

-

K_m (mM)

118 ± 7

-

-

167 ± 11

-

-

-

-

-

k_cat/K_m (mM^-1 min^-1)

2.8 ± 0.2

-

-

1.1 ± 0.1

-

-

-

-

-

Xylobiose

k_cat (min^-1)

449 ± 5

(446 ± 9)

764 ± 17

536 ± 6

(670 ± 50)

-

-

(332 ± 11)

(115 ± 7)

K_m (mM)

0.099 ± 0.005

(10.3 ± 0.3)

7.0 ± 0.3

0.43 ± 0.02

(48 ± 4)

-

-

(11 ± 1)

(23 ± 2)

k_cat/K_m (mM^-1 min^-1)

4,500 ± 200

(43 ± 2)

109 ± 5

1,250 ± 60

(14 ± 2)

-

-

(30 ± 2)

(5.0 ± 0.5)

Xylotriose

k_cat (min^-1)

524 ± 10

(590 ± 30)

920 ± 40

620 ± 20

(1,500 ± 300)

-

-

(260 ± 30)

(270 ± 30)

K_m (mM)

0.10 ± 0.01

(11 ± 1)

6.1 ± 0.5

0.34 ± 0.04

(100 ± 20)

-

-

(10 ± 2)

(41 ± 6)

k_cat/K_m (mM^-1 min^-1)

5,100 ± 400

(54 ± 6)

150 ± 14

1,800 ± 200

(15 ± 5)

-

-

(27 ± 5)

(7 ± 1)