Biomasses of Penicillium camemberti, Penicillium roqueforti and Geotrichum candidum strains were grown and harvested after a culture incubation period that corresponded to the maximal dry weight of mycelium as well as to lipoxygenase (LOX) activity. The crude enzymatic extracts were recovered from the homogenized mycelium cells and the partially purified LOX extracts were obtained by ammonium sulfate precipitation. Using linoleic acid as substrate, the partially purified LOX extracts from P. camemberti, P. roqueforti and G. candidum exhibited a major activity, at pH 6.50, 5.50 and 3.75, respectively, and a minor one at pH 8.00. The partially purified LOX extracts exhibited an overall preferential specificity towards free fatty acids, including linoleic, linolenic and arachidonic acids, than that for fatty acid acylglycerols, including mono-, di- and tri-linolein. The Km and Vmax values for partially purified LOXs were investigated. Normal phase high-performance liquid chromatography (NP-HPLC) and gas-liquid chromatography/mass spectrometry (GC/MS) analyses showed that the LOX activity of the partially purified LOX extracts converted mainly linoleic acid into the corresponding 9- and 13-hydroperoxides (HPODs); however, the production of a relatively important proportion of 10-HPOD, ranging from 4 to 9% of the total HPODs, was also demonstrated with the partially purified LOX extracts from Penicillium sp. The chiral phase HPLC analysis demonstrated the production, by the partially purified LOX extracts from Penicillium sp., of both (R)- and (S)-enantiomers of HPODs, with a slightly higher proportion of the (S)-enantiomers. The crude enzymatic extracts from Penicillium sp. were incubated, at pH 6.50, with individual racemic mixture of 9(R,S)-, 10(R,S)-, 12( R,S)- and 13(R,S)-HPODs, prepared by photooxidation and separated by NP-HPLC. The experimental results indicated that only 10( S)-HPOD isomer was cleaved by a hydroperoxide lyase activity that resulted by the for
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.36678 |
| Date | January 2000 |
| Creators | Perraud, Xavier. |
| Contributors | Kermasha, S. (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 | Doctor of Philosophy (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: 001764649, proquestno: NQ64642, Theses scanned by UMI/ProQuest. |
Page generated in 0.0013 seconds