Catalytic Stability of S-1-(4-Hydroxyphenyl)-Ethanol Dehydrogenase from Aromatoleum aromaticum
Abstract
:1. Introduction
2. Results and Discussion
2.1. Synthesis of Chiral Alcohols with Pure S-HPED
2.2. Effects of pH and Reaction Temperature on the Activity of S-HPED
2.2.1. Impact of Glucose
2.2.2. Thermal Shift Assay
2.3. Catalytic Stability and Aggregation of S-HPED at Elevated Temperatures
2.3.1. Catalytic Stability Study under Storage Conditions
2.3.2. Catalytic Stability Study under Reactor Conditions
2.3.3. Aggregation under Storage Conditions
2.4. Modelling of Thermal Inactivation under Storage Conditions
3. Materials and Methods
3.1. Chemicals
3.2. Preparation of S-HPED
3.3. Synthesis of Chiral Alcohols with Pure S-HPED
3.4. Chiral Chromatographic Analysis
3.5. Effect of pH on the Initial Activity of S-HPED and Activity Decrease Rate
3.6. Effects of Temperature on the Activity of S-HPED
3.7. UV-Vis Activity Assay
3.8. S-HPED Stability Tests under Reaction Condition
3.9. Chromatographic Analyses—Reverse Phase (RP-HPLC)
3.10. Aggregation Measurements
3.11. Dynamic Light Scattering
3.12. Thermal Shift Assay
3.13. Thermal Inactivation Experiments
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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No. | Substrate | Product | Product S (%) | Product R (%) | % ee | Relative Activity * (%) |
---|---|---|---|---|---|---|
0 | acetophenone | 1-phenylethanol | 100 | 0 | 100 | 100 |
1 | 4′-chloroacetophenone | 1-(4-chlorophenyl)ethanol | 100 | 0 | 100 | 123 |
2 | 4′-metoxyacetophenone | 1-(4-methoxyphenyl)ethanol | 100 | 0 | 100 | 82 |
3 | 4′-fluoroacetophenone | 1-(4-fluorophenyl)ethanol | 100 | 0 | 100 | 108 |
4 | 4′-nitroacetophenone | 1-(4-nitrophenyl)ethanol | - | - | 100 | 201 |
5 | 3′-aminoacetophenone | 1-(3-aminophenyl)ethanol | 100 | 0 | 100 | 57 |
5 | 4′-aminoacetophenone | 1-(4-aminophenyl)ethanol | 96 | 4 | 92 | 7 |
7 | 4′-ethylacetophenone | 1-(4-ethylphenyl)ethanol | - | - | 100 | 70 |
8 | 4′-bromoacetophenone | 1-(4-bromophenyl)ethanol | 100 | 0 | 100 | 130 |
9 | 4-acetylbenzonitrile | 4-(1-hydroxyethyl)benzonitrile | - | - | 100 | 186 |
10 | 2,2-dichloroacetophenone | 2,2-dichloro-1-phenylethanol | - | - | 100 | 344 |
11 | 4′-acetylbiphenyl | 1-(biphenyl-4-yl)ethanol | - | - | 100 | 204 |
12 | 1-indanone | 1-indanol | - | - | - | 0 |
13 | 2,3-benzofuran | 2,3-dihydro-1-benzofuran-3-ol | - | - | - | 0 |
14 | 3-phenyl-1-indanone | 3-phenyl-1-indanol | - | - | - | 0 |
15 | 6-hydroxy-1-indanone | 2,3-dihydro-1H-indene-1,6-diol | - | - | - | 0 |
Time | pH 5.5 | pH 6.5 | pH 7.5 | pH 8.3 | pH 9.0 | |
---|---|---|---|---|---|---|
without additives | 0 h | 100% | 100% | 100% | 100% | 100% |
0.5 h | 0.3% | 3.2% | 2.4% | 2.5% | 2% | |
with 2 mM acetophenone | 0 h | 100% | 100% | 100% | 100% | 100% |
0.5 h | 1.4% | 2.6% | 2.4% | 2.1% | 1.2% | |
with 1.5 M glucose | 0 h | 100% | 100% | 100% | 100% | 100% |
0.5 h | 9.3% | 62.5% | 38% | 48% | 42.3% | |
2 h | 3.5% | 10% | 8.7% | 9.8% | 15.6% | |
5 h | 2% | 5.4% | 3% | 2.1% | 9.3% |
Temperature (°C) | Glucose-Free | in 1.5 M Glucose | ||||
---|---|---|---|---|---|---|
k (min−1) | sso | SD (%) | k (min−1) | sso | SD (%) | |
25 | 0.00583 ± 0.00022 | 0.01418 | 3.303 | --- | --- | --- |
30 | 0.01489 ± 0.00053 | 0.01373 | 3.132 | --- | --- | --- |
35 | 0.04029 ± 0.00144 | 0.01639 | 3.305 | --- | --- | --- |
40 | 0.10868 ± 0.00301 | 0.00563 | 2.082 | 0.00315 ± 0.00011 | 0.01119 | 3.19 |
45 | 0.36106 ± 0.01189 | 0.00155 | 1.488 | 0.00867 ± 0.00026 | 0.01694 | 3.36 |
50 | --- | --- | --- | 0.03558 ± 0.00134 | 0.01179 | 3.274 |
55 | --- | --- | --- | 0.12340 ± 0.00354 | 0.00661 | 2.451 |
60 | --- | --- | --- | 0.36083 ± 0.00673 | 0.00296 | 1.57 |
Parameters/ Inactivationconditions | k0 (min−1) | Ea (kJ mol−1) | SD (%) | n |
---|---|---|---|---|
glucose-free | 0.04266 ± 0.00127 | 156.60 ± 4.46 | 3.27 | 65 |
in 1.5 M glucose | 0.03447 ± 0.00127 | 211.55 ± 4.63 | 3.78 | 63 |
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Tataruch, M.; Illeová, V.; Kluza, A.; Cabadaj, P.; Polakovič, M. Catalytic Stability of S-1-(4-Hydroxyphenyl)-Ethanol Dehydrogenase from Aromatoleum aromaticum. Int. J. Mol. Sci. 2024, 25, 7385. https://doi.org/10.3390/ijms25137385
Tataruch M, Illeová V, Kluza A, Cabadaj P, Polakovič M. Catalytic Stability of S-1-(4-Hydroxyphenyl)-Ethanol Dehydrogenase from Aromatoleum aromaticum. International Journal of Molecular Sciences. 2024; 25(13):7385. https://doi.org/10.3390/ijms25137385
Chicago/Turabian StyleTataruch, Mateusz, Viera Illeová, Anna Kluza, Patrik Cabadaj, and Milan Polakovič. 2024. "Catalytic Stability of S-1-(4-Hydroxyphenyl)-Ethanol Dehydrogenase from Aromatoleum aromaticum" International Journal of Molecular Sciences 25, no. 13: 7385. https://doi.org/10.3390/ijms25137385