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

Abstract

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.

Microbial Proteases Applications

“…Microbial proteases are the most commercially exploited enzyme worldwide. A large number of intracellular proteases are produced by microbes playing a vital role in differentiation, protein turnover, hormone regulation, and cellular protein pool, whereas extracellular proteases are significant in protein hydrolysis (Rao et al, 1998; Johnvesly and Naik, 2001; Adrio and Demain, 2014), such as in processing of photographic film (Kumar and Takagi, 1999; Patil and Chaudhari, 2009), enzymatic synthesis on the basis of solvent and detergent preparation (Simkhada et al, 2010a), substrate specificity (Soroor et al, 2009), thermal tolerance (Amoozegar et al, 2007), and production of zein hydrolysates (Miyaji et al, 2006; Dodia et al, 2008; Jaouadi et al, 2008).…”

Purification and Characterization of a Small Thermostable Protease from Streptomyces sp. CNXK100

“…P3 ( Cheng et al 2015 ). Besides, Mn 2+ and Fe 2+ induced a partial reduction in the activity of the CNXK100 protease whereas both were observed to completely abolish the protease activity from S. olivochromogenes ( Simkhada et al 2010 ). This observation underscores the potential applicability of the enzyme under conditions characterized by the presence of these heavy metals.…”

“…The K M value serves as an indicator of the enzyme’s affinity for its substrate. The K M of the CNXK100 protease was 1.3–4.2 times higher than that of the metalloprotease from S. olivochromogenes (0.74 mg/ml) ( Simkhada et al 2010 ) and the alkaline serine protease from S. griseus (2.50 mg/ml) ( El-Khonezy et al 2015 ). However, the V max value of the CNXK100 protease, representing the maximum reaction rate of substrate conversion, was remarkably higher (846–17200 times) than those of the protease from S. olivochromogenes (3876 U/mg) ( Simkhada et al 2010 ) and S. griseus (190 U/mg) ( El-Khonezy et al 2015 ).…”

Potential of Protease from Bacillus species for Biomedical and Industrial Applications

“…Microbial proteases are the most fiscally abused protein all throughout the planet. A colossal number of intracellular proteases are made by microorganisms accepting an essential occupation in detachment, protein turnover, chemical rule, and cell protein pool, while extracellular proteases are vital in protein hydrolysis (Adrio and Demain, 2014, for instance, in treatment of photographic film (Kumar and Takagi, 1999;Patil and Chaudhari, 2009), enzymatic combination dependent on dissolvable and cleaning agent status (Simkhada et al, 2010a), substrate disposition (Soroor et al, 2009), warm flexibility (Amoozegar et al, 2007), and production of zein hydrolysates (Miyaji et al, 2006;Dodia et al, 2008;Jaouadi et al, 2008).…”