Purification and partial characterisation of camel milk xanthine oxidoreductase.
Xanthine oxidoreductase (XOR) was purified in the presence of dithiothrietol from camel milk with yields of up to 22.2mg/l that were comparable to those obtained from bovine and human milk sources. On SDS-PAGE, the freshly purified camel milk XOR had a protein flavin (A280/A450) ratio of 5.3 +/- 0.4 and appeared homogenous with a single major band of approximately Mr 145.3 KDa. Surprisingly, in all the batches (n = 8) purified camel milk XOR showed no detectable activity towards xanthine or NADH. The molybdenum content of camel XOR was comparable to human and goat milk enzymes. After resulphuration, camel milk XOR gave a specific activity of 1.1 nmol/min/mg and 13.0 nmol/min/mg enzyme towards pterin (fluorimetric assay) and xanthine (spectrophotometric assay) respectively. This activity was markedly lower than that of human, bovine and goat enzymes obtained under the same conditions. These findings suggest that the molybdo-form of camel enzyme is totally under desulpho inactive form. It is possible that camel neonates are equipped with an enzymic system that reactivates XOR in their gut and consequently generates antibacterial reactive oxygen species.
Baghiani
A
A
Laboratory of Applied Biochemistry, Faculty of Sciences, University of Setif, Algeria.
Harrison
R
R
Benboubetra
M
M
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002162:Camelus; D002417:Cattle; D004229:Dithiothreitol / Q000494:pharmacology; D005260:Female; D006801:Humans; D008892:Milk / Q000201:enzymology*; D008970:Molecular Weight; D008982:Molybdenum / Q000032:analysis / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D010088:Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism; D011622:Pterins / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000494:pharmacology; D019820:Xanthine / Q000378:metabolism; D014968:Xanthine Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D014969:Xanthine Oxidase / Q000302:isolation & purification* / Q000378:metabolism
D000900:Anti-Bacterial Agents; D011622:Pterins; D017382:Reactive Oxygen Species; D009243:NAD; D019820:Xanthine; D008982:Molybdenum; D010088:Oxidoreductases; D014968:Xanthine Dehydrogenase; D014969:Xanthine Oxidase; D004229:Dithiothreitol