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An Oxidant- and Organic Solvent-Resistant Alkaline Metalloprotease from Streptomyces olivochromogenes

Simkhada, Jaya Ram, Cho, Seung Sik, Park, Seong Ju, Mander, Poonam, Choi, Yun Hee, Lee, Hyo Jeong, Yoo, Jin Cheol
Applied biochemistry and biotechnology 2010 v.162 no.5 pp. 1457-1470
EDTA (chelating agent), Streptomyces olivochromogenes, agarose, calcium, cobalt, copper, detergents, ethylene, filtration, gelatin, hydrogen peroxide, ion exchange, iron, manganese, metalloproteinases, molecular weight, oxidants, pH, polyacrylamide gel electrophoresis, solvents, thermal stability, zinc
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 °C and stable between pH 7.0 and 10.0. SOMP was stable below 45 °C and Ca²⁺ increased its thermostability. Ca²⁺ enhanced while Co²⁺, Cu²⁺, Zn²⁺, Mn²⁺, and Fe²⁺ inhibited the activity. Ethylenediaminetetraacetic acid and ethylene glycol-bis (β-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 H₂O₂. 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.