A Noyel Alcohol Resistant Metalloproteinase , Purification and

named yimelysin, was purified from the cutture supernatant by three column chromatographies. About 150mg of purified vime]ysin -'as obtained from 3.3 Iiters of the culture supernatant with a high yield of 57'ZF. The purified vimelysin showed a single protein band on SDS-PAGE with molecular vveight of 38,OOO. The isoelectric point of vimel)'sin was 4.3 by isoelectric focusing. The optimum pH of vimelysin was pH 8.0 or pH 6.5 using casein or furylacrylo}'1-glyeyl-leucine amide (FAGLA) as substrates, respectively. The optimum temperature of s'imet)'sin was 500C when casein was used as a substrate, but it was 150C when FAGI.A was used as a substrate. Interestingty, vimelysin activity was completely retained after 48h of incubation at 25eC in the presence of 50% ethanol. Moreover, vimel)'sin showed 40% activit)' of the control eyen in the presence of 10(X) ethanol, while thermol)'sin showed enly 5`M, actiyity under the same conditions.