Xylanase production from a Bacillus subtilis gene cloned into a strain of Escherichia coli was measured. Although this gene was expressed in E. coli at several temperatures, efficient normal xylanase secretion did not occur, the observed protein release apparently depending on cell leakage or lysis. Screening for a better microbial protein secretor free of cellulase selected B. cereus #259. The strain had wild plasmids that were hard to eliminate using acridine orange and elevated temperature curing techniques. While still bearing 5 wild plasmids, attempts to transform B. cereus #259 were unsuccessful using conventional methods and electroporation. Another strain, B. cereus #518, found to be free of wild plasmids, was then used. A bidirectional vector shuttle plasmid (pMK3) was employed to carry the cloned gene into this B. cereus strain. Transformation was carried out by high voltage electroporation. Xylanase production by the new B. cereus clone was similar to that from E. coli, but was shown to be continuously and normally secreted. The xylanase gene products from the E. coli and B. cereus hosts were shown to function identically. Both xylanases improved the delignification of unbleached softwood and hardwood kraft pulps, thus reducing the Cl$ sb2$ required to achieve a given degree of bleaching, without altering the physical properties of the fibers. Using a target kappa number lignin content) of 5, xylanase pretreatment of aspen kraft pulp led to a 22% saving of chlorine. Adsorbable organic halogens in the bleachery effluent were also lowered by more than 50%.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.69517 |
| Date | January 1993 |
| Creators | Tremblay, Louis |
| Contributors | Archibald, Fred (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 Microbiology.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001338489, proquestno: AAIMM87810, Theses scanned by UMI/ProQuest. |
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