Pfaendtner, JimMundy, ChristopherWaggett, Ava2024-09-092024-09-092024Waggett_washington_0250O_26712.pdfhttps://hdl.handle.net/1773/51826Thesis (Master's)--University of Washington, 2024Microbial lipases constitute a class of biocatalysts with the ability to cleave ester linkages of long-chain triglycerides, a property which makes them particularly attractive for industrial applications ranging from food processing to pharmaceutical preparation. Candida rugosa lipase (CRL) is among the enzymes most frequently used in biotransformation. A notable feature of CRL is its propensity for interfacial activation, exhibiting elevated catalytic rates when acting at the interface between aqueous and hydrophobic phases. This phenomenon can be understood in the context of the enzyme’s hydrophobic substrate and the presence of a mobile lid domain. To advance our understanding of interfacial activation, we explore the dynamics of CRL rotation at the octane-water interface in this work. To do so, we employ Molecular Dynamics to evaluate the free energy of rotation at the interface, identifying a global minimum at the point of lid alignment to the interface that coincides with and is further stabilized by lid opening. Additionally, we investigate the role of leucine residues outside of the lid domain as they serve to instigate rotation of the lid towards the interface. These findings serve to further elucidate the mechanism of CRL activation at the oil-water interface and point towards a holistic effect of rotation and lid opening that is mediated by both lid and non-lid surface residues.application/pdfen-USnoneAdsorptionEnzymesHydrophobicityInterfacesLipasesMolecular dynamicsComputational chemistryBiophysicsMolecular chemistryChemical engineeringThesis