A novel alcohol resistant metalloproteinase, vimelysin, from vibrio sp. T1800: purification and characterization.
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 culture supernatant by three column chromatographies. About 150 mg 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 focusing. 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 degrees C when casein was used as a substrate, but it was 15 degrees C when FAGLA was used as a substrate. Interestingly, vimelysin activity was completely retained after 48 h of incubation at 25 degrees 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.
Oda
K
K
Department of Applied Biology, Kyoto Institute of Technology, Japan.
Okayama
K
K
Okutomi
K
K
Shimada
M
M
Sato
R
R
Takahashi
S
S
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008666:Metalloendopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D012997:Solvents; D013696:Temperature; D013820:Thermolysin / Q000737:chemistry / Q000378:metabolism; D014733:Vibrio / Q000201:enzymology*
D001426:Bacterial Proteins; D011480:Protease Inhibitors; D012997:Solvents; D008666:Metalloendopeptidases; C099068:vimelysin; D013820:Thermolysin