The enzyme chitin deacetylase was extracted from the fungus Mucor rouxii. The fungus was grown aerobically at pH 4.5, and a temperature of 28 +/- 2°C, using a shaker speed of 100 rpm. The highest enzymatic activity was obtained at 36 h of Mucor rouxii's growth. The protein content of Mucor rouxii was a function of incubation time and it increased in the course of the fungal growth up to 3 days. / The enzyme was partial purified by ammonium sulfate fractionation, followed by ion exchange chromatography using a Fast Protein Liquid Chromatography (FPLC) System. The specific activity in the 60--85% ammonium sulfate fraction showed higher specific activity (51.44 U/mg) and a 2.2-fold purification as compared with that of the crude extract (24.27 U/mg). The fraction after ion exchange chromatography step showed a specific activity of 233.65 U/mg, representing about 7.2-fold purification compared to the crude extract. This ion exchange fraction showed protein bands with molecular weights ranging from 45 to 75 kDa on SDS-PAGE (12%). / The enzyme showed optimal activity at a temperature of 50°C, and more than 90% of its activity was retained after 30 min incubation at this temperature. The optimal pH activity of chitin deacetylase from Mucor rouxii ranged from 5.5--6. The enzyme retained more than 80% of its activity in the pH range of 4.5--8.5 after 1 h incubation at room temperature, suggesting that this enzyme is quite stable in the slightly acidic to alkaline pH range. However, a lower pH (<4) and a higher pH (>9) caused the enzyme to lose more than 60% of its activity. / It was shown that the chitin deacetylase did not require the following metal ions (Co+2 and Mn+2) for activity. However, Mn+2 slightly increased the enzyme activity. The activity of the enzyme was also inhibited by EDTA.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.79132 |
| Date | January 2003 |
| Creators | Soltani, Sahel |
| Contributors | Simpson, B. K. (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 Food Science and Agricultural Chemistry.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001985463, proquestno: AAIMQ88301, Theses scanned by UMI/ProQuest. |
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