Tuning Immobilized Enzyme Features by Combining Solid-Phase Physicochemical Modification and Mineralization
Abstract
:1. Introduction
2. Results and Discussion
2.1. Preparation of the Immobilized and Chemically Modified TLL-Biocatalysts
2.2. Modification of Octyl-TLL with Different Phosphate Salts
2.3. Effect of Mineralization on the Immobilized Enzyme Previously Modified
2.4. Preparation of the Immobilized and Chemically Modified CALB-Biocatalysts
2.5. Mineralization of Octyl-CALB
2.6. Effect of Mineralization on the Immobilized Enzyme Previously Modified
3. Materials and Methods
3.1. Materials
3.2. Methods
3.2.1. Immobilization of Lipases on Octyl-Agarose Beads
3.2.2. Immobilization of Lipases on Octyl-Agarose Beads
3.2.3. Modification of Immobilized Enzyme with Metallic Salt/Phosphate
3.2.4. Thermal Inactivation of Different Lipase Biocatalysts
3.2.5. Determination of the Biocatalysts Activities versus Different Substrates
Hydrolysis of p-NPB
Hydrolysis of Triacetin
Hydrolysis of R- or S-Methyl Mandelate
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Biocatalysts | Activity (U/g) | |||
---|---|---|---|---|
p-NPB | Triacetin | R-Methyl Mandelate | S-Methyl Mandelate | |
Octyl-TLL | 1416 ± 51 | 601 ± 6 | 1.2 ± 0.1 | 3.2 ± 0.2 |
Octyl-TLL-GA | 1350 ± 91 | 48 ± 2 | 0.8 ± 0.1 | 2.4 ± 0.1 |
Octyl-TLL-Amin | 1675 ± 49 | 88 ± 4 | 0.6 ± 0.1 | 3.2 ± 0.2 |
Octyl-TLL-TNBS | 1764 ± 82 | 60 ± 3 | 0.9 ± 0.1 | 3.1 ± 0.2 |
Octyl-TLL-PEI | 1363 ± 61 | 90 ± 4 | 9.8 ± 0.5 | 7.7 ± 0.4 |
Octyl-TLL-DS | 985 ± 52 | 28 ± 1 | 7.5 ± 0.4 | 6.5 ± 0.2 |
Biocatalysts | Activity (U/g) | |||
---|---|---|---|---|
p-NPB | Triacetin | R-Methyl Mandelate | S-Methyl Mandelate | |
Octyl-TLL | 1416 ± 51 | 61 ± 6 | 1.2 ± 0.1 | 3.2 ± 0.2 |
Octyl-TLL-ZnP * | 1040 ± 64 | 88 ± 6 | 0.9 ± 0.1 | 3.0 ± 0.1 |
Octyl-TLL-CoP | 1207 ± 72 | 65 ± 5 | 1.1 ± 0.1 | 2.9 ± 0.2 |
Octyl-TLL-CuP | 1144 ± 90 | 67 ± 6 | 0.8 ± 0.1 | 2.7 ± 0.1 |
Octyl-TLL-NiP | 1371 ± 87 | 53 ± 3 | 1.5 ± 0.1 | 3.0 ± 0.2 |
Octyl-TLL-MgP | 1665 ± 41 | 66 ± 3 | 1.5 ± 0.1 | 2.4 ± 0.1 |
Line | Biocatalysts | Activity (U/g) | |||
---|---|---|---|---|---|
1 | p-NPB | Triacetin | R-Methyl Mandelate | S-Methyl Mandelate | |
2 | Octyl-TLL-GA | 1350 ± 91 | 48 ± 2 | 0.8 ± 0.1 | 2.4 ± 0.1 |
3 | Octyl-TLL-GA-ZnP | 1408 ± 84 | 50 ± 2 | 0.7 ± 0.1 | 3.0 ± 0.1 |
4 | Octyl-TLL-GA-CoP | 1133 ± 61 | 35 ± 2 | 0.2 ± 0.1 | 2.0 ± 0.1 |
5 | Octyl-TLL-GA-CuP | 1142 ± 42 | 38 ± 2 | 0.6 ± 0.1 | 2.1 ± 0.1 |
6 | Octyl-TLL-GA-NiP | 1308 ± 54 | 48 ± 2 | 0.6 ± 0.1 | 2.2 ± 0.1 |
7 | Octyl-TLL-GA-MgP | 1404 ± 120 | 46 ± 2 | 0.7 ± 0.1 | 2.4 ± 0.1 |
8 | Octyl-TLL-Amin | 1675 ± 49 | 88 ± 4 | 0.6 ± 0.1 | 3.2 ± 0.2 |
9 | Octyl-TLL-Amin-ZnP | 1422 ± 208 | 78 ± 4 | 0.6 ± 0.1 | 3.1 ± 0.2 |
10 | Octyl-TLL-Amin-CoP | 1647 ± 87 | 53 ± 3 | 0.1 ± 0.1 | 2.2 ± 0.1 |
11 | Octyl-TLL-Amin-CuP | 1618 ± 36 | 79 ± 4 | 0.9 ± 0.4 | 3.0 ± 0.2 |
12 | Octyl-TLL-Amin-NiP | 1863 ± 107 | 63 ± 3 | 0.6 ± 0.1 | 2.8 ± 0.1 |
13 | Octyl-TLL-Amin-MgP | 1979 ± 90 | 79 ± 4 | 0.8 ± 0.1 | 2.9 ± 0.1 |
14 | Octyl-TLL-TNBS | 1763 ± 82 | 60 ± 3 | 0.9 ± 0.1 | 3.1 ± 0.2 |
15 | Octyl-TLL-TNBS-ZnP | 1836 ± 97 | 47 ± 2. | 0.9 ± 0.1 | 2.7 ± 0.1 |
16 | Octyl-TLL-TNBS-CoP | 1591 ± 109 | 46 ± 2 | 0.4 ± 0.1 | 2.4 ± 0.1 |
17 | Octyl-TLL-TNBS-CuP | 1501 ± 76 | 45 ± 2 | 1.0 ± 0. 14 | 3.0 ± 0.2 |
18 | Octyl-TLL-TNBS-NiP | 1592 ± 128 | 44 ± 2 | 1.4 ± 0.1 | 3.1 ± 0.2 |
19 | Octyl-TLL-TNBS-MgP | 1544 ± 60 | 46 ± 2 | 1.1 ± 0.1 | 3.0 ± 0.2 |
Line | Biocatalysts | Activity (U/g) | |||
---|---|---|---|---|---|
1 | p-NPB | Triacetin | R-Methyl Mandelate | S-Methyl Mandelate | |
2 | Octyl-TLL-PEI | 1363 ± 61 | 90 ± 4 | 9.8 ± 0.5 | 7.7 ± 0.4 |
3 | Octyl-TLL-PEI-ZnP | 1479 ± 20 | 106 ± 5 | 8.2 ± 0.4 | 8.2 ± 0.5 |
4 | Octyl-TLL-PEI-CoP | 1442 ± 31 | 108 ± 6 | 7.7 ± 0.4 | 8.1 ± 0.3 |
5 | Octyl-TLL-PEI-CuP | 1351 ± 80 | 120 ± 6 | 8.7 ± 0.4 | 8.0 ± 0.2 |
6 | Octyl-TLL-PEI-NiP | 1309 ± 72 | 94 ± 4 | 8.5 ± 0.5 | 7.9 ± 0.3 |
7 | Octyl-TLL-PEI-MgP | 1323 ± 50 | 80 ± 5 | 8.2 ± 0.4 | 7.7 ± 0.3 |
8 | Octyl-TLL-DS | 985 ± 52 | 28 ± 1 | 7.5 ± 0.4 | 6.5 ± 0.2 |
9 | Octyl-TLL-DS-ZnP | 1041 ± 31 | 37 ± 2 | 9.9 ± 0.4 | 7.3 ± 0.4 |
10 | Octyl-TLL-DS-CoP | 1084 ± 46 | 27 ± 1 | 8.1 ± 0.3 | 6.1 ± 0.2 |
11 | Octyl-TLL-DS-CuP | 898 ± 54 | 30 ± 2 | 8.9 ± 0.4 | 7.2 ± 0.4 |
12 | Octyl-TLL-DS-NiP | 1041 ± 40 | 28 ± 1 | 8.5 ± 0.5 | 7.2 ± 0.3 |
13 | Octyl-TLL-DS-MgP | 951 ± 23 | 31 ± 1 | 8.9 ± 0.4 | 7.4 ± 0.4 |
Biocatalysts | Activity (U/g) | |||
---|---|---|---|---|
p-NPB | Triacetin | R-Methyl Mandelate | S-Methyl Mandelate | |
Octyl-CALB | 1151 ± 62 | 132 ± 6 | 19.5 ± 0.9 | 39.0 ± 1.9 |
Octyl-CALB-GA | 1325 ± 23 | 94 ± 5 | 16.8 ± 0.9 | 28.0 ± 1.4 |
Octyl-CALB-Amin | 1312 ± 36 | 77 ± 3 | 26.5 ± 1.3 | 36.3 ± 1.9 |
Octyl-CALB-TNBS | 1336 ± 36 | 89 ± 7 | 18.3 ± 0.8 | 41.1 ± 2.0 |
Octyl-CALB-PEI | 1518 ± 91 | 143 ± 9 | 26.1 ± 1.5 | 33.3 ± 1.7 |
Octyl-CALB-DS | 1294 ± 85 | 121 ± 4 | 20.0 ± 0.8 | 43.2 ± 2.2 |
Biocatalysts | Activity (U/g) | |||
---|---|---|---|---|
p-NPB | Triacetin | R-Methyl Mandelate | S-Methyl Mandelate | |
Octyl-CALB | 1151 ± 62 | 132 ± 6 | 19.5 ± 0.9 | 39.0 ± 1.9 |
Octyl-CALB-ZnP | 1172 ± 43 | 194 ± 7 | 25.6 ± 1.3 | 49.5 ± 2.9 |
Octyl-CALB-CoP | 967 ± 52 | 220 ± 13 | 27.7 ± 1.4 | 50.2 ± 2.2 |
Octyl-CALB-CuP | 998 ± 73 | 270 ± 14 | 26.8 ± 1.3 | 50.1 ± 2.9 |
Octyl-CALB-NiP | 1218 ± 78 | 222 ± 13 | 20.4 ± 1.0 | 43.5 ± 2.1 |
Octyl-CALB-MgP | 1141 ± 49 | 82 ± 5 | 17.5 ± 0.9 | 29.5 ± 1.6 |
Line | Biocatalysts | Activity (U/g) | |||
---|---|---|---|---|---|
1 | p-NPB | Triacetin | R-Methyl Mandelate | S-Methyl Mandelate | |
2 | Octyl-CALB-GA | 1324 ± 23 | 94 ± 5 | 16.8 ± 0.9 | 28.0 ± 1.4 |
3 | Octyl-CALB-GA-ZnP | 1195 ± 23 | 92 ± 5 | 9.7 ± 0.5 | 25.0 ± 1.9 |
4 | Octyl-CALB-GA-CoP | 1324 ± 39 | 85 ± 4 | 15.5 ± 0.8 | 26.6 ± 1.3 |
5 | Octyl-CALB-GA-CuP | 1195 ± 29 | 82 ± 4 | 14.1 ± 0.7 | 31.4 ± 1.4 |
6 | Octyl-CALB-GA-NiP | 1263 ± 44 | 78 ± 2 | 18. 9 ± 0.8 | 27.8 ± 1.2 |
7 | Octyl-CALB-GA-MgP | 1264 ± 56 | 121 ± 6 | 23.8 ± 1.0 | 29.2 ± 1.6 |
8 | Octyl-CALB-Amin | 1312 ± 36 | 77 ± 3 | 26.5 ± 1.3 | 36.3 ± 1.9 |
9 | Octyl-CALB-Amin-ZnP | 1266 ± 21 | 112 ± 7 | 23.6 ± 1.4 | 27.9 ± 1.3 |
10 | Octyl-CALB-Amin-CoP | 1225 ± 69 | 122 ± 6 | 25.3 ± 1.1 | 27.0 ± 1.0 |
11 | Octyl-CALB-Amin-CuP | 1341 ± 63 | 97 ± 6 | 24.3 ± 0.9 | 27.6 ± 1.2 |
12 | Octyl-CALB-Amin-NiP | 1369 ± 102 | 116 ± 6 | 24.7 ± 1.4 | 37.0 ± 1.9 |
13 | Octyl-CALB-Amin-MgP | 1264 ± 56 | 121 ± 6 | 23.8 ± 1.0 | 29.2 ± 1.6 |
14 | Octyl-CALB-TNBS | 1336 ± 36 | 89 ± 7 | 18.3 ± 0.8 | 41.1 ± 2.0 |
15 | Octyl-CALB-TNBS-ZnP | 1266 ± 70 | 139 ± 7 | 16.7 ± 0.8 | 39.1 ± 2.0 |
16 | Octyl-CALB-TNBS-CoP | 1263 ± 81 | 140 ± 8 | 17.4 ± 0.7 | 37.5 ± 1.9 |
17 | Octyl-CALB-TNBS-CuP | 1128 ± 58 | 169 ± 8 | 19.3 ± 0.9 | 43.3 ± 2.1 |
18 | Octyl-CALB-TNBS-NiP | 1214 ± 119 | 153 ± 7 | 20.0 ± 0.8 | 38.2 ± 1.9 |
19 | Octyl-CALB-TNBS-MgP | 1170 ± 34.9 | 145 ± 9 | 18.6 ± 1.2 | 39.8 ± 2.0 |
Line | Biocatalysts | Activity (U/g) | |||
---|---|---|---|---|---|
1 | p-NPB | Triacetin | R-Methyl Mandelate | S-Methyl Mandelate | |
2 | Octyl-CALB-PEI | 1518 ± 91 | 143 ± 9 | 26.1 ± 1.5 | 33.3 ± 1.7 |
3 | Octyl-CALB-PEI-ZnP | 1769 ± 30 | 181 ± 7 | 26.9 ± 1.4 | 38.2 ± 2.0 |
4 | Octyl-CALB-PEI-CoP | 1638 ± 33 | 201 ± 11 | 26.3 ± 1.4 | 40.8 ± 2.1 |
5 | Octyl-CALB-PEI-CuP | 1656 ± 48 | 219 ± 14 | 17.2 ± 0.9 | 45.9 ± 2.9 |
6 | Octyl-CALB-PEI-NiP | 1489 ± 93 | 200 ± 11 | 12.5 ± 0.7 | 47.8 ± 2.3 |
7 | Octyl-CALB-PEI-MgP | 1468 ± 89 | 207 ± 13 | 19.1 ± 1.0 | 48.5 ± 2.6 |
8 | Octyl-CALB-DS | 1294 ± 85 | 121 ± 4 | 20.0 ± 0.8 | 43.2 ± 2.2 |
9 | Octyl-CALB-DS-ZnP | 1251 ± 51 | 132 ± 8 | 23.6 ± 1.1 | 43.2 ± 1.9 |
10 | Octyl-CALB-DS-CoP | 1248 ± 57 | 144 ± 8 | 21.4 ± 1.0 | 44.8 ± 2.6 |
11 | Octyl-CALB-DS-CuP | 1188 ± 101 | 182 ± 10 | 20.9 ± 0.8 | 46.6 ± 2.5 |
12 | Octyl-CALB-DS-NiP | 1232 ± 31 | 159 ± 9 | 19.3 ± 0.6 | 41.0 ± 1.9 |
13 | Octyl-CALB-DS-MgP | 1308 ± 27 | 179 ± 10 | 21.5 ± 0.9 | 44.3 ± 1.4 |
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Guimarães, J.R.; Carballares, D.; Rocha-Martin, J.; Tardioli, P.W.; Fernandez-Lafuente, R. Tuning Immobilized Enzyme Features by Combining Solid-Phase Physicochemical Modification and Mineralization. Int. J. Mol. Sci. 2022, 23, 12808. https://doi.org/10.3390/ijms232112808
Guimarães JR, Carballares D, Rocha-Martin J, Tardioli PW, Fernandez-Lafuente R. Tuning Immobilized Enzyme Features by Combining Solid-Phase Physicochemical Modification and Mineralization. International Journal of Molecular Sciences. 2022; 23(21):12808. https://doi.org/10.3390/ijms232112808
Chicago/Turabian StyleGuimarães, José R., Diego Carballares, Javier Rocha-Martin, Paulo W. Tardioli, and Roberto Fernandez-Lafuente. 2022. "Tuning Immobilized Enzyme Features by Combining Solid-Phase Physicochemical Modification and Mineralization" International Journal of Molecular Sciences 23, no. 21: 12808. https://doi.org/10.3390/ijms232112808