Laccase and Its Mutant Displayed on the <i>Bacillus subtilis</i> Spore Coat for Oxidation of Phenolic Compounds in Organic Solvents
Enzymes displayed on the <i>Bacillus subtilis</i> spore coat have several features that are useful for biocatalysis. The enzyme is preimmobilized on an inert surface of the spore coat, which is due to the natural sporulation process. As a result, protein stability can be increased, and t...
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MDPI AG
2021-05-01
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| Online Access: | https://www.mdpi.com/2073-4344/11/5/606 |
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| author | Silu Sheng Edgardo T. Farinas |
| author_facet | Silu Sheng Edgardo T. Farinas |
| author_sort | Silu Sheng |
| collection | DOAJ |
| description | Enzymes displayed on the <i>Bacillus subtilis</i> spore coat have several features that are useful for biocatalysis. The enzyme is preimmobilized on an inert surface of the spore coat, which is due to the natural sporulation process. As a result, protein stability can be increased, and they are resistant to environmental changes. Next, they would not lyse under extreme conditions, such as in organic solvents. Furthermore, they can be easily removed from the reaction solution and reused. The laboratory evolved CotA laccase variant T480A-CotA was used to oxidize the following phenolic substrates: (+)-catechin, (−)-epicatechin, and sinapic acid. The kinetic parameters were determined and T480A-CotA had a greater <i>V</i><sub>max</sub>/<i>K</i><sub>m</sub> than wt-CotA for all substrates. The <i>V</i><sub>max</sub>/<i>K</i><sub>m</sub> for T480A-CotA was 4.1, 5.6, and 1.4-fold greater than wt-CotA for (+)-catechin, (−)-epicatechin, and sinapic acid, respectively. The activity of wt-CotA and T480A-CotA was measured at different concentrations from 0–70% in organic solvents (dimethyl sulfoxide, ethanol, methanol, and acetonitrile). The <i>V</i><sub>max</sub> for T480A-CotA was observed to be greater than the wt-CotA in all organic solvents. Finally, the T480A-CotA was recycled 7 times over a 23-h period and up to 60% activity for (+)-catechin remained. The product yield was up to 3.1-fold greater than the wild-type. |
| first_indexed | 2024-03-10T11:36:09Z |
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| language | English |
| last_indexed | 2024-03-10T11:36:09Z |
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| spelling | doaj.art-f6f01b72b46448c8b44da43f83411f432023-11-21T18:55:36ZengMDPI AGCatalysts2073-43442021-05-0111560610.3390/catal11050606Laccase and Its Mutant Displayed on the <i>Bacillus subtilis</i> Spore Coat for Oxidation of Phenolic Compounds in Organic SolventsSilu Sheng0Edgardo T. Farinas1GenScript USA Inc., 860 Centennial Avenue, Piscataway, NJ 08854, USADepartment of Chemistry and Environmental Science, New Jersey Institute of Technology, University Heights, Newark, NJ 07102, USAEnzymes displayed on the <i>Bacillus subtilis</i> spore coat have several features that are useful for biocatalysis. The enzyme is preimmobilized on an inert surface of the spore coat, which is due to the natural sporulation process. As a result, protein stability can be increased, and they are resistant to environmental changes. Next, they would not lyse under extreme conditions, such as in organic solvents. Furthermore, they can be easily removed from the reaction solution and reused. The laboratory evolved CotA laccase variant T480A-CotA was used to oxidize the following phenolic substrates: (+)-catechin, (−)-epicatechin, and sinapic acid. The kinetic parameters were determined and T480A-CotA had a greater <i>V</i><sub>max</sub>/<i>K</i><sub>m</sub> than wt-CotA for all substrates. The <i>V</i><sub>max</sub>/<i>K</i><sub>m</sub> for T480A-CotA was 4.1, 5.6, and 1.4-fold greater than wt-CotA for (+)-catechin, (−)-epicatechin, and sinapic acid, respectively. The activity of wt-CotA and T480A-CotA was measured at different concentrations from 0–70% in organic solvents (dimethyl sulfoxide, ethanol, methanol, and acetonitrile). The <i>V</i><sub>max</sub> for T480A-CotA was observed to be greater than the wt-CotA in all organic solvents. Finally, the T480A-CotA was recycled 7 times over a 23-h period and up to 60% activity for (+)-catechin remained. The product yield was up to 3.1-fold greater than the wild-type.https://www.mdpi.com/2073-4344/11/5/606protein displayprotein engineeringlaccaseorganic solvent stability |
| spellingShingle | Silu Sheng Edgardo T. Farinas Laccase and Its Mutant Displayed on the <i>Bacillus subtilis</i> Spore Coat for Oxidation of Phenolic Compounds in Organic Solvents Catalysts protein display protein engineering laccase organic solvent stability |
| title | Laccase and Its Mutant Displayed on the <i>Bacillus subtilis</i> Spore Coat for Oxidation of Phenolic Compounds in Organic Solvents |
| title_full | Laccase and Its Mutant Displayed on the <i>Bacillus subtilis</i> Spore Coat for Oxidation of Phenolic Compounds in Organic Solvents |
| title_fullStr | Laccase and Its Mutant Displayed on the <i>Bacillus subtilis</i> Spore Coat for Oxidation of Phenolic Compounds in Organic Solvents |
| title_full_unstemmed | Laccase and Its Mutant Displayed on the <i>Bacillus subtilis</i> Spore Coat for Oxidation of Phenolic Compounds in Organic Solvents |
| title_short | Laccase and Its Mutant Displayed on the <i>Bacillus subtilis</i> Spore Coat for Oxidation of Phenolic Compounds in Organic Solvents |
| title_sort | laccase and its mutant displayed on the i bacillus subtilis i spore coat for oxidation of phenolic compounds in organic solvents |
| topic | protein display protein engineering laccase organic solvent stability |
| url | https://www.mdpi.com/2073-4344/11/5/606 |
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