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dc.contributor.advisorBrandsdal, Bjørn Olav
dc.contributor.advisorIsaksen, Geir Villy
dc.contributor.authorBurkow, Magnus
dc.date.accessioned2020-10-29T08:18:48Z
dc.date.available2020-10-29T08:18:48Z
dc.date.issued2020-06-01
dc.description.abstractDrug discovery and refinement for use in the industry is an important and lucrative field within chemistry. In this regard, computational chemistry has shown to be a valuable tool in assisting drug discovery. A set of intriguing proteins used in drug discovery are cold-adapted enzymes. These enzymes have a lower activation point which makes them of interest in industrial use. In this study, the cold-adapted shrimp alkaline phosphatase’s (SAP) affinity towards the immunoassay substrates PNPP and AMPPD has been studied. With free energy calculations using the linear interaction energy (LIE) and free energy perturbation (FEP) methods, SAP’s affinity towards the substrates has been determined. In addition, putative mutations of SAP have been carried out by modelling SAP upon the already catalytic effective calf intestinal alkaline phosphatase (CIP) to increase its affinity towards PNPP and AMPPD. It was found that both substrates bound favourably to SAP and all its different mutations. Specificity towards PNPP and AMPPD changed depending on the mutations, where G102R would bind PNPP over AMPPD, while H149D would bind AMPPD over PNPP. The most successful mutation was a metal exchange of Zn2+ in the M3 metal site to Mg2+ according to the LIE method. The FEP method on the other hand revealed a decrease in binding free energies when transforming Zn2+ to Mg2+. One possible solution given is that a key water molecule coordinated poorly in the FEP simulations compared to LIE simulations. However, even though literature shows that Mg2+ increases the catalytic effect, this does not necessarily imply that the proteins affinity towards the substrates need to increase.en_US
dc.identifier.urihttps://hdl.handle.net/10037/19697
dc.language.isoengen_US
dc.publisherUiT Norges arktiske universiteten_US
dc.publisherUiT The Arctic University of Norwayen_US
dc.rights.accessRightsopenAccessen_US
dc.rights.holderCopyright 2020 The Author(s)
dc.rights.urihttps://creativecommons.org/licenses/by-nc-sa/4.0en_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)en_US
dc.subject.courseIDKJE-3900
dc.subjectVDP::Matematikk og Naturvitenskap: 400::Kjemi: 440::Teoretisk kjemi, kvantekjemi: 444en_US
dc.subjectVDP::Mathematics and natural science: 400::Chemistry: 440::Theoretical chemistry, quantum chemistry: 444en_US
dc.titleComputational Studies of Selected Mutants of Shrimp Alkaline Phosphataseen_US
dc.typeMaster thesisen_US
dc.typeMastergradsoppgaveen_US


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Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)