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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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