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Table 3 Stability Characteristics of HRP Immobilisation Mutants in free solution and immobilised on CNBr-activated Sepharose

From: Arginine-to-lysine substitutions influence recombinant horseradish peroxidase stability and immobilisation effectiveness

 

Thermal Stability

Solvent and H2O2 Stability

Mutant

T50 (°C)

k-value (min-1)

SD (min-1)

t1/2 (min)

DMSO C50

MeOH C50

DMF C50

H2O2 C50

Free Solution

        

WT

50

0.0559

± 0.0034

12

35

53

14

17

Mutant 1

51

0.0535

± 0.0005

12

34

44

12

38

Mutant 2

50

0.0568

± 0.0057

12

30

35

16

44

Mutant 3

48

0.0109

± 0.0007

6

35

45

25

17

Mutant 4

50

0.0445

± 0.0003

17

30

35

16

33

Immobilised

        

WT

-

0.0255

± 0.0023

27

32

40

14

6

Mutant 1

-

0.0223

± 0.0020

3

24

32

17

5

Mutant 2

-

0.0544

± 0.0044

13

28

38

10

5

Mutant 3

-

0.0152

± 0.0017

5

42

43

29

5

Mutant 4

-

0.0491

± 0.0049

14

28

38

15

5

  1. WT, Wildtype; Mutant 1, R118K/R159K/R283K; Mutant 2, R118K/R159K; Mutant 3, R118K/R159K/K232N/K241F/R283K; Mutant 4, R118K/R159K/K232N/K241F. Modelled k-values were calculated using the Enzfitter™ software package (Version 1.05. Cambridge UK: Biosoft Ltd.; 1987). Values are the mean of three independent determinations in each case. Standard deviations were < 5% for solvent and < 10% for H2O2 studies. Units of C50 are % v/v for solvents and mM for H2O2.
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BMC Biotechnology

ISSN: 1472-6750

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