Abstract
Given that lipase is an enzyme applicable in various industrial fields and water-miscible organic solvents are important reaction media for developing industrial-scale biocatalysis, a structure-based strategy was explored to stabilize lipase A from Bacillus subtilis in a water–ethanol cosolvent. Site-directed mutagenesis of ethanol-interacting sites resulted in 4 mutants, i.e., Ser16Gly, Ala38Gly, Ala38Thr, and Leu108Asn, which were stable in 50% ethanol and had up to 1.8-fold higher stability than the wild-type. In addition, Leu108Asn was more thermostable at 45 °C than the wild type. The results discussed in this study not only provide insights into strategies for enzyme engineering to improve organic solvent resistance but also suggest perspectives on pioneering routes for constructing enzyme-based biorefineries to produce value-added fuels and chemicals.
Original language | English |
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Article number | 125394 |
Journal | Bioresource Technology |
Volume | 337 |
DOIs | |
State | Published - Oct 2021 |
Bibliographical note
Publisher Copyright:© 2021 Elsevier Ltd
Keywords
- Bacillus subtilis lipase A
- Engineering ethanol-interacting residues
- Rational surface design
- Stability in water–ethanol cosolvent