Cold-Active Lipase-Based Biocatalysts for Silymarin Valorization through Biocatalytic Acylation of Silybin
Abstract
:1. Introduction
2. Results and Discussion
2.1. Evaluation of Psychrobacter SC65A.3 Lipolytic Activity on Different Substrates
2.2. Characterization of the Immobilized Cold-Active Lipase
2.3. Cold-Active Lipase Catalyzing Silybin Acylation
3. Materials and Methods
3.1. Chemicals and Solutions
3.2. Plate Screening Assays
3.3. Preparation of the Protein Extract
3.4. Enzyme Assay
3.5. Lipase Immobilization
3.5.1. Preparation of the Immobilized Lipase Biocatalyst
3.5.2. Characterization of the Immobilized Lipase Biocatalyst
- mi—initial protein amount (after concentration of the extracellular extract);
- ms—protein amount in the supernatant after immobilization step;
- LAimmob—lipase activity of the immobilized protein fraction;
- LAi—lipase activity of the solution used for immobilization;
- LAs—lipase activity of the supernatant after immobilization.
3.6. Biocatalytic System for Silybin Acylation
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Recovery of Lipase Activity (%) | Immobilization Yield (%) | Protein Loading (%) | |
---|---|---|---|
S1 | 82 ± 5.1 | 80 ± 4.5 | 99 ± 5.3 |
S2 | 128 ± 6.3 | 84 ± 4.7 | 99 ± 4.8 |
S3 | 80 ± 5.4 | 76 ± 4.3 | 93 ± 4.6 |
S4 | 82 ± 5.5 | 74 ± 4.8 | 96 + 4.9 |
S5 | 105 ± 5.1 | 89 ± 4.0 | 90 ± 4.3 |
Biocatalyst | Km (mM) | kcat (min−1) | kcat/Km (mM−1 min−1) | |||
---|---|---|---|---|---|---|
25 °C | 37 °C | 25 °C | 37 °C | 25 °C | 37 °C | |
Free | 2.18 ± 0.15 | 1.85 ± 0.09 | 1.63 ± 0.08 | 1.95 ± 0.09 | 0.75 ± 0.05 | 1.05 ± 0.09 |
S1 | 1.61 ± 0.08 | 1.17 ± 0.09 | 0.99 ± 0.05 | 0.97 ± 0.06 | 0.62 ± 0.04 | 0.83 ± 0.04 |
S2 | 0.56 ± 0.04 | 0.65 ± 0.03 | 0.93 ± 0.05 | 0.99 ± 0.06 | 1.67 ± 0.09 | 1.54 ± 0.07 |
S3 | 4.18 ± 0.21 | 1.41 ± 0.07 | 2.29 ± 0.09 | 1.25 ± 0.07 | 0.55 ± 0.03 | 0.88 ± 0.04 |
S4 | 0.63 ± 0.03 | 0.78 ± 0.05 | 0.65 ± 0.03 | 0.92 ± 0.05 | 1.02 ± 0.09 | 1.17 ± 0.09 |
S5 | 0.94 ± 0.05 | 0.64 ± 0.03 | 0.85 ± 0.08 | 1.2 ± 0.09 | 0.90 ± 0.05 | 1.88 ± 0.09 |
Support | Immobilization Method | |
---|---|---|
S1 | sMP-COOH | EDC |
S2 | fMP-COOH | EDC |
S3 | sMP-NH2 | EDC |
S4 | sMP-NH-NH2 | NaIO4 |
S5 | MTC-NH2 | GA |
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Gheorghita, G.R.; Paun, V.I.; Neagu, S.; Maria, G.-M.; Enache, M.; Purcarea, C.; Parvulescu, V.I.; Tudorache, M. Cold-Active Lipase-Based Biocatalysts for Silymarin Valorization through Biocatalytic Acylation of Silybin. Catalysts 2021, 11, 1390. https://0-doi-org.brum.beds.ac.uk/10.3390/catal11111390
Gheorghita GR, Paun VI, Neagu S, Maria G-M, Enache M, Purcarea C, Parvulescu VI, Tudorache M. Cold-Active Lipase-Based Biocatalysts for Silymarin Valorization through Biocatalytic Acylation of Silybin. Catalysts. 2021; 11(11):1390. https://0-doi-org.brum.beds.ac.uk/10.3390/catal11111390
Chicago/Turabian StyleGheorghita, Giulia Roxana, Victoria Ioana Paun, Simona Neagu, Gabriel-Mihai Maria, Madalin Enache, Cristina Purcarea, Vasile I. Parvulescu, and Madalina Tudorache. 2021. "Cold-Active Lipase-Based Biocatalysts for Silymarin Valorization through Biocatalytic Acylation of Silybin" Catalysts 11, no. 11: 1390. https://0-doi-org.brum.beds.ac.uk/10.3390/catal11111390