An oxidant- and organic solvent-resistant alkaline metalloprotease from Streptomyces olivochromogenes.
Organic solvent- and detergent-resistant proteases are important from an industrial viewpoint. However, they have been less frequently reported and only few of them are from actinomycetes. A metalloprotease from Streptomyces olivochromogenes (SOMP) was purified by ion exchange with Poros HQ and gel filtration with Sepharose CL-6B. Apparent molecular mass of the enzyme was estimated to be 51 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gelatin zymography. The activity was optimum at pH 7.5 and 50 degrees C and stable between pH 7.0 and 10.0. SOMP was stable below 45 degrees C and Ca(2+) increased its thermostability. Ca(2+) enhanced while Co(2+), Cu(2+), Zn(2+), Mn(2+), and Fe(2+) inhibited the activity. Ethylenediaminetetraacetic acid and ethylene glycol-bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, but not phenylmethylsulfonyl fluoride, aprotinin, and pefabloc SC, significantly suppressed the activity, suggesting that it might be a metalloprotease. Importantly, it is highly resistant against various detergents, organic solvents, and oxidizing agents, and the activity is enhanced by H(2)O(2). The enzyme could be a novel protease based on its origin and peculiar biochemical properties. It may be useful in biotechnological applications especially for organic solvent-based enzymatic synthesis.
Simkhada
Jaya Ram
JR
Department of Pharmacy, College of Pharmacy, Chosun University, Dong-gu, Gwangju 501-759, Korea.
Cho
Seung Sik
SS
Park
Seong Ju
SJ
Mander
Poonam
P
Choi
Yun Hee
YH
Lee
Hyo Jeong
HJ
Yoo
Jin Cheol
JC
D000468:Alkalies / Q000378:metabolism*; D002850:Chromatography, Gel; D003902:Detergents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007700:Kinetics; D045726:Metalloproteases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D009930:Organic Chemicals / Q000494:pharmacology*; D016877:Oxidants / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000494:pharmacology; D019163:Reducing Agents / Q000494:pharmacology; D012997:Solvents / Q000494:pharmacology*; D013302:Streptomyces / Q000187:drug effects / Q000201:enzymology*; D013696:Temperature
D000468:Alkalies; D003902:Detergents; D008670:Metals; D009930:Organic Chemicals; D016877:Oxidants; D011480:Protease Inhibitors; D019163:Reducing Agents; D012997:Solvents; D045726:Metalloproteases