Conversion of Racemic Unnatural Amino Acids to Optically Pure Forms by a Coupled Enzymatic Reaction
Abstract
:1. Introduction
2. Results and Discussion
2.1. Metal-Chelating Amino Acids in GCE Technology
2.2. System Design for the Preparation of Optically Pure BPA and HQA
2.3. RgDAAO and TtAT Activity for BPA and HQA
2.4. Enzymatic Preparation of Optically Pure BPA and HQA
2.5. Application of Optically Pure BPA and HQA to GCE Experiments
3. Materials and Methods
3.1. Expression and Purification of RgDAAO
3.2. Expression and Purification of TtAT
3.3. RgDAAO Reaction
3.4. TtAT Reaction
3.5. Coupled Reaction of RgDAAO and TtAT
3.6. Application to the GCE Technique
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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Substrate | kcat (min−1) | Km (mM) | kcat/Km (min−1mM−1) |
---|---|---|---|
NPA | 1710 ± 85 | 0.052 ± 0.0087 | 3.3 × 104 |
BPA | 448 ± 20 | 0.087 ± 0.010 [b] | 0.52 × 104 |
HQA | 124 ± 0.4 | 0.043 ± 0.0094 [b] | 0.29 × 104 |
Substrate | Conversion (%) [a] | Enantiomeric Excess (%) | |
---|---|---|---|
RgDAAO Reaction | Coupled Reaction | ||
BPA | 97 | 95 | 95 |
HQA | 98 | 98 | 98 |
Samples | GFP Intensity | |||
---|---|---|---|---|
BPA | HQA | |||
500 μM | 2000 μM | 250 μM | 1000 μM | |
l-form [a] | 1983 | 3937 | 2456 | 3456 |
d,l-form [b] | 1059 (1942) [c] | 2885 (3211) [c] | 1633 (2326) [c] | 2868 (2036) [c] |
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Lee, H.; Kim, D.; Kim, S.; Lee, H.S. Conversion of Racemic Unnatural Amino Acids to Optically Pure Forms by a Coupled Enzymatic Reaction. Molecules 2021, 26, 1274. https://doi.org/10.3390/molecules26051274
Lee H, Kim D, Kim S, Lee HS. Conversion of Racemic Unnatural Amino Acids to Optically Pure Forms by a Coupled Enzymatic Reaction. Molecules. 2021; 26(5):1274. https://doi.org/10.3390/molecules26051274
Chicago/Turabian StyleLee, Hannae, Dongchan Kim, Sooin Kim, and Hyun Soo Lee. 2021. "Conversion of Racemic Unnatural Amino Acids to Optically Pure Forms by a Coupled Enzymatic Reaction" Molecules 26, no. 5: 1274. https://doi.org/10.3390/molecules26051274