Isolation, Purification and Characterization of new cold active subtilisin-like protease from Bacillus sp. strain EL-GU1

Abstract

Proteases are hydrolytic enzymes that slice peptide bonds between amino acid residues and these enzymes have various industrial applications including detergent, food, pharmaceutical, leather and diagnostic reagent industries. Among them, alkaline proteases, the most commercialized enzymes in the industry, are of particular interest due to their potential applications in the detergent industry as cleaning additives. In this study, a novel alkaline protease from Bacillus sp. strain EL-GU1 was reported showing highest activity at pH 6 and 25°C. The novel protease was purified by using ammonium sulfate precipitation and identified by 16S rDNA sequencing. Highest activity was observed as 3300 µmol/min^-1mg^-1 when casein used as a substrate. Kinetic parameters of the enzyme were determined; K_M, V_max, k_cat and catalytic efficiency values were calculated as 1.4 mM, 1 mM/s, 2.10^-7 s^-1, 0.14 10^-7 s^-1M^-1, respectively. These results indicated that the novel cold active protease from Bacillus sp. strain EL- GU1 can be a good candidate for the detergent industry

Keywords

• Bacillus sp.,protease,enzyme characterization,cold active and alkali protease

Soğuk Aktif ve Alkali Subtilisin Benzeri Proteaz Enziminin Bacillus sp. strain EL-GU1’ den İzolasyonu, Saflaştırılması ve Karekterizasyonu

Abstract

One of the important hydrolytic enzymes are proteases that slice peptide bonds between amino acid residues. Proteases have various industrial applications including detergent, food, pharmaceutical, leather and diagnostic reagent industries. Among them, the most commercialized enzymes are alkaline proteases in the industry. Due to their potential applications in the detergent industry as cleaning additives, they are of particular interest. In this study, a novel protease from Bacillus sp. strain EL-GU1 was reported showing highest activity at pH 6 and 20°C. The novel protease was purified by using ammonium sulfate precipitation and identified by 16S rDNA sequencing. Highest activity was observed as 3300 µmol/min^-1mg^-1 when casein used as a substrate. Kinetic parameters of the enzyme were determined; K_M, V_max, k_cat and catalytic efficiency values were calculated as 1.4 mM, 1 mM/s, 2.10^-7 s^-1, 0.14 10^-7 s^-1M^-1, respectively. These results indicated that the novel cold active protease from Bacillus sp. strain EL- GU1 can be a good candidate for the detergent industry.

 

Keywords

• Bacillus sp.,proteaz,enzim karakterizasyonu,soğuk aktif ve alkali proteaz

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