Enzyme immobilization strategies for the synthesis of perdeuterated molecules
(2018) KBKM05 20181Pure and Applied Biochemistry
Computational Chemistry
- Abstract
- This thesis covers established procedures for the successful and attempted immobilization of lipases and phospholipases to various hydrophobic support materials, utilizing a rotating bed reactor, magnetic stirrer and tube-rotator. The design of a robust platform was established with great instrumental knowledge and proper conditioning for the creation of efficient immobilizations and activity assays. Influencing factors as substrate amount, pH, temperature, solvent selection, solubility, RPM, Co-factors, support material, protein loading and water activity have all been considered and assessed, although not thoroughly evaluated due to the purpose of solely finding an efficient and working system for immobilization. The success of the... (More)
- This thesis covers established procedures for the successful and attempted immobilization of lipases and phospholipases to various hydrophobic support materials, utilizing a rotating bed reactor, magnetic stirrer and tube-rotator. The design of a robust platform was established with great instrumental knowledge and proper conditioning for the creation of efficient immobilizations and activity assays. Influencing factors as substrate amount, pH, temperature, solvent selection, solubility, RPM, Co-factors, support material, protein loading and water activity have all been considered and assessed, although not thoroughly evaluated due to the purpose of solely finding an efficient and working system for immobilization. The success of the immobilization experiments has primarily been validated utilizing the NanoDrop- instrument, for protein concentration estimations, and a variety of activity assays. The activity assays are performed utilizing both already commercially immobilized lipases and self-immobilized (in-house) lipases and phospholipases. The conversion rate, established from the activity assays were analyzed utilizing Gas Chromatography Flame Ionization Detector (GC-FID), Nuclear Magnetic Resonance (NMR) and Thin Layer Chromatography (TLC). Lipase from Rhizopus oryzae (ROL) was successfully immobilized, with the proper conditions and working procedures, via adsorption to various hydrophobic support materials. ROL effected a 94% conversion of the lauric acid in the esterification reaction of lauric acid to propyl laurate, and a maintained catalytic activity of 99% after two cycles. No confirmed immobilization of the sn2- specific phospholipase A2 from Porcine pancreas (PLA2) could be presented during the thesis, although a deeper knowledge for the reaction settings and conditions were acquired, resulting in promising opportunities for future development. (Less)
- Popular Abstract (Swedish)
- Användandet av nya strategier för immobilisering av enzymer (proteiner) möjliggör
för stora framsteg inom neutron forskning. Genom noggranna tester upprättades väl
valda villkor för användandet av enzymer, med ambitionen att effektivisera
framställning av så kallade ”tunga molekyler”. Något som i sin tur är värdefullt för att förstå hur celler är strukturerade samt signalerar och interagerar med varandra.
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/8951126
- author
- Bogojevic, Oliver LU
- supervisor
- organization
- course
- KBKM05 20181
- year
- 2018
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Applied Chemistry, Lipases, Phospholipases, Rhizopus oryzae
- language
- English
- id
- 8951126
- date added to LUP
- 2022-06-30 10:50:41
- date last changed
- 2022-06-30 10:50:41
@misc{8951126, abstract = {{This thesis covers established procedures for the successful and attempted immobilization of lipases and phospholipases to various hydrophobic support materials, utilizing a rotating bed reactor, magnetic stirrer and tube-rotator. The design of a robust platform was established with great instrumental knowledge and proper conditioning for the creation of efficient immobilizations and activity assays. Influencing factors as substrate amount, pH, temperature, solvent selection, solubility, RPM, Co-factors, support material, protein loading and water activity have all been considered and assessed, although not thoroughly evaluated due to the purpose of solely finding an efficient and working system for immobilization. The success of the immobilization experiments has primarily been validated utilizing the NanoDrop- instrument, for protein concentration estimations, and a variety of activity assays. The activity assays are performed utilizing both already commercially immobilized lipases and self-immobilized (in-house) lipases and phospholipases. The conversion rate, established from the activity assays were analyzed utilizing Gas Chromatography Flame Ionization Detector (GC-FID), Nuclear Magnetic Resonance (NMR) and Thin Layer Chromatography (TLC). Lipase from Rhizopus oryzae (ROL) was successfully immobilized, with the proper conditions and working procedures, via adsorption to various hydrophobic support materials. ROL effected a 94% conversion of the lauric acid in the esterification reaction of lauric acid to propyl laurate, and a maintained catalytic activity of 99% after two cycles. No confirmed immobilization of the sn2- specific phospholipase A2 from Porcine pancreas (PLA2) could be presented during the thesis, although a deeper knowledge for the reaction settings and conditions were acquired, resulting in promising opportunities for future development.}}, author = {{Bogojevic, Oliver}}, language = {{eng}}, note = {{Student Paper}}, title = {{Enzyme immobilization strategies for the synthesis of perdeuterated molecules}}, year = {{2018}}, }