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AMB Express

Table 1 Specificity constants of LinBUT, LinB MI , and their intermediate mutants

From: Stepwise enhancement of catalytic performance of haloalkane dehalogenase LinB towards β-hexachlorocyclohexane

Enzyme

Position of the different amino acid residues

Specificity constantkcat/Km(M-1s-1)

Data source

Folda

Mutant name

81

112

134

135

138

247

253

HCH→PCHL

% to LinBMI

PCHL→TCDL

% to LinBMI

 

0

LinBUT

 

A

A

I

A

I

A

M

17 ± 0

9

2.0 ± 1

0.2

This studyb

1

LinBUT A81T

M1-1

T

A

I

A

I

A

M

92 ± 3

50

2.0 ± 1

0.2

This study

1

LinBUT A112V

M1-2

A

V

I

A

I

A

M

4 ± 0

2.1

0

0

This study

1

LinBUT I134V

M1-3

A

A

V

A

I

A

M

21 ± 0

11

5 ± 1

0.5

This studyb

1

LinBUT A135T

M1-4

A

A

I

T

I

A

M

23 ± 1

12

6 ± 2

0.6

This study

1

LinBUT I138L

M1-5

A

A

I

A

L

A

M

35 ± 1

18

5 ± 1

0.5

This study

1

LinBUT A247H

M1-6

A

A

I

A

I

H

M

68 ± 2

36

4 ± 1

0.4

This studyb

1

LinBUT M253I

M1-7

A

A

I

A

I

A

I

39 ± 1

21

4 ± 1

0.4

This study

2

LinBUT I134V/A247H

M2-1

A

A

V

A

I

H

M

30 ± 0

16

7 ± 1

0.7

This studyb

3

LinBUT A112V/ I134V/A247H

M3-1

A

V

V

A

I

H

M

53 ± 3

28

17 ± 2

1.7

This study

3

LinBUT I134V/I138L/A247H

M3-2

A

A

V

A

L

H

M

69 ± 10

36

31 ± 7

3.1

This study

3

LinBUT I134V/A247H/M253I

M3-3

A

A

V

A

I

H

I

33 ± 1

17

7 ± 1

0.7

This study

4

LinBUT A112V/I134V/I138L/A247H

M4-1

A

V

V

A

L

H

M

48 ± 4

25

85 ± 10

8.5

This study

4

LinBUT I134V/I138L/A247H/M253I

M4-2

A

A

V

A

L

H

I

1.0 ± 0.2 x 102

53

83 ± 20

8.3

This study

5

LinBUT A112V/I134V/I138L/A247H/M253I

M5-1

A

V

V

A

L

H

I

1.2 ± 0.1 x 102

63

1.3 ± 0.2 x 102

13

This study

5

LinBUT A81T/I134V/I138L/A247H/M253I

M5-2

T

A

V

A

L

H

I

79 ± 10

42

50 ± 10

5

This study

5

LinBUT I134V/A135T/I138L/A247H/M253I

M5-3

A

A

V

T

L

H

I

68 ± 8

36

90 ± 20

9

This study

5

LinBUT A81T/A112V/I134V/I138L/A247H

M5-4

T

V

V

A

L

H

M

91 ± 9

48

83 ± 10

8.3

This study

5

LinBUT A112V/I134V/A135T/I138L/A247H

M5-5

A

V

V

T

L

H

M

84 ± 20

44

73 ± 20

7.3

This study

6

LinBMI T81A

M6-1

A

V

V

T

L

H

I

68 ± 3

36

9.5 ± 5.0 x 102

95

Okai et al. [2013]

6

LinBMI V112A

M6-2

T

A

V

T

L

H

I

1.0 ± 0.09 x 102

53

2.3 ± 0.4 x 102

23

Okai et al. [2013]

6

LinBMI V134I

M6-3

T

V

I

T

L

H

I

1.2 ± 0.05 x 102

63

80 ± 3

8

Ito et al. [2007]

6

LinBMI T135A

M6-4

T

V

V

A

L

H

I

83 ± 5

44

4.2 ± 1 x 102

42

Okai et al. [2013]

6

LinBMI L138I

M6-5

T

V

V

T

I

H

I

1.3 ± 0.1 x 102

68

2.2 ± 0.4 x 102

22

Okai et al. [2013]

6

LinBMI H247A

M6-6

T

V

V

T

L

A

I

2.1 ± 0.2 x 102

111

2.4 ± 0.2 x 102

24

Ito et al. [2007]

6

LinBMI I253M

M6-7

T

V

V

T

L

H

M

2.1 ± 0.3 x 102

111

1.4 ± 0.2 x 102

14

Okai et al. [2013]

7

LinBMI

 

T

V

V

T

L

H

I

1.9 ± 0.08 x 102

100

1.0 ± 0.3 x 103

100

Okai et al. [2013]

  1. aNumber of mutation into LinBUT.
  2. bThe activities of LinBUT, M1-3, M1-6, and M2-1 have already been measured in our previous study (Ito et al. [2007]), but they were reanalyzed in this study for the critical comparison with other mutants.
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