Usage of GD-95 and GD-66 lipases as fusion partners leading to improved chimeric enzyme LipGD95-GD66
| Autor: | Renata Gudiukaite, Ausra Veteikyte, Audrius Gegeckas, Egle Lastauskiene, Inga Matijosyte, Mikas Sadauskas, Gytis Druteika, Vilius Malunavicius |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine food.ingredient Recombinant Fusion Proteins Argan oil Protein Engineering 01 natural sciences Biochemistry 03 medical and health sciences chemistry.chemical_compound food Structural Biology 010608 biotechnology Catalytic Domain Enzyme Stability Organic chemistry Lipase Enzyme Inhibitors Molecular Biology Ethanol biology Esterification Substrate (chemistry) Geobacillus General Medicine Transesterification Hydrogen-Ion Concentration Kinetics 030104 developmental biology chemistry Biocatalysis Metals biology.protein Solvents Methanol Acyl group |
| Zdroj: | International journal of biological macromolecules. 118 |
| ISSN: | 1879-0003 |
| Popis: | Lipases are used as biocatalysts in industrial processes mainly because of their stability at broad temperature and pH range, resistance to organic solvents and wide spectrum of substrates. The usage of several lipolytic domains, each with different activity and resistance profiles, enables both the flexibility and efficiency of industrial processes. In this study, GD-95 and GD-66 lipases produced by Geobacillus sp. 95 and Geobacillus sp. 66, respectively, were used as fusion partners to create a new fused lipolytic enzyme LipGD95-GD66. Chimeric LipGD95-GD66 lipase displayed tenfold increase in activity (200 U/mg) compared to parental GD-66 lipase, improved Vmax (10 μmol/min mg−1) and catalytic efficiency (2 ∗ 105 min−1 mM−1) for p-NP palmitate as a substrate and increased activity at 70–75 °C compared to both parental lipases. All three lipases also retained >50% of their lipolytic activity after incubation with methanol, n-hexane, ethanol and DMF for longer than three weeks, highlighting a great prospect for application in industrial processes. Moreover, transesterification results revealed the capability of parental GD-95 lipase to be the most promising biocatalyst for production of methyl and ethyl esters through eco-friendly transesterification using argan oil and ethanol/methanol as acceptors of acyl group. |
| Databáze: | OpenAIRE |
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