A Novel Alcohol Resistant Metalloproteinase, Vimelysin, from Vibrio sp. T1800: Purification and Characterization

Abstract

We found a novel metalloproteinase, which has high activity at low temperatures and in the presence of organic solvents, in the culture supernatant of a marine bacterium, Vibrio sp. T1800. The metalloproteinase, named vimelysin, was purified from the dulture supernatant by three column chromatographies. About 150mg of purified vimelysin was obtained from 3.3 liters of the culture supernatant with a high yield of 57%. The purified vimelysin showed a single protein band on SDS-PAGE with molecular weight of 38, 000. The isoelectric point of vimelysin was 4.3 by isoelectric f6cusing. The optimum pH of vimelysin was pH 8.0 or pH 6.5 using casein or furylacryloyl-glycyl-leucine amide (FAGLA) as substrates, respectively. The optimum temperature of vimelysin was 50°C when cdsein was uSed as a substrate, but it was 15°C when FAGLA was used as a substrate. Interestingly, vimelysin activity was completely retained after 48h of incubation at 25°C in the presence of 50% ethanol. Moreover, vimelysin showed 40% activity of the control even in the presence of 10% ethanol, while thermolysin showed only 5% activity under the same conditions.