In order to make use of enzymes in unnatural applications more efficiently, novel enzymes have to be developed and they may arise from an existing enzyme with similar performance. The fast way to improve the performance of the enzyme is by directed evolution. It alters a naturally existing protein, without in depth knowledge of the starting enzyme and produces mutants containing the required activities. / The lipase from Bacillus thermocatenulatus is an alko-thermophilic enzyme. Optimization of the lipase production is carried out for use with the new screening method using a pH indicator to evaluate hydrolytic activities. This newly cloned lipase is next characterized with two substrate libraries made up of commercially available general and enantiomerically pure esters. An important chiral synthon solketal is identified as the directed evolution target molecule for this lipase. By modifying with error-prone PCR, the enantioselectivity of the lipase towards solketal n-octanoate is improved.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.30691 |
| Date | January 1999 |
| Creators | Liu, Andrew Man Fai, 1975- |
| Contributors | Kazlauskas, Romas J. (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Chemistry.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001744491, proquestno: MQ64395, Theses scanned by UMI/ProQuest. |
Page generated in 0.0014 seconds