Partial Purification of Protease by A Novel Bacterium, Bacillus cereus and Enzymatic Properties ​

Abstract

Proteases E.C. 3.4.X.X catalyze the hydrolysis of peptide bonds in proteins. Microbial proteases are classified based on their mode of action and biocatalytic mechanisms. They participate in most aspects of cell nutrition, physiology, and regulation, and in microbial pathogenesis. Protease production was detected from 15 bacteria isolated from soil samples and the one showed the highest protease activity was selected. The strain was identified and determined as Bacillus cereus by 16S rRNA phylogenetic analysis. After optimization of protease production from the novel medium, the Michaelis-Menten kinetics was studied. Temperature, pH and, time parameters of protease incubation was determined and maximum temperature was detected at 50oC as 5.15 IU/ml. The optimum pH range of the enzyme was in between pH 7-9. The crude enzyme was approximately 2-fold purified by dialysis.

Keywords

• Protease,Production,Optimization,Bacillus cereus

Yeni İzole Bacillus cereus Proteazının Kısmi Saflaştırılması ve Enzimolojik Özellikleri

Abstract

Proteazlar E.C. 3.4.X.X proteinlerdeki peptid bağlarının hidrolizinden sorumlu enzimlerdir. Mikrobiyal proteazlar, etkide bulundukları bölge ve biyokatalitik mekanizmalarına göre sınıflandırılırlar. Bu proteazlar hücre beslenmesi, fizyolojisi, düzenlenmesi ve mikrobiyal patogenezde rol oynamaktadırlar. Toprak örneklerinden izole edilmiş 15 bakterinin proteaz aktiviteleri saptanmış ve en yüksek aktiviteye sahip suş seçilerek stoklanmıştır. Bu suşun 16S rRNA filogenetik analizi yapılarak Bacillus cereus olduğu saptanmıştır. Proteaz üretiminin optimizasyonu sonrasında bu suşun Michaelis-Menten kinetiği hesaplanmıştır. Proteaz inkübasyonunun sıcaklık, pH ve zaman parametreleri araştırılmış ve maksimum sıcaklık aktivitesi 50oC’de 5.15 IU/ml olarak saptanmıştır. Enzimin optimum pH aralığı 7-9’dur. Ham enzim, diyaliz yöntemiyle yaklaşık olarak 2 kat saflaştırılmıştır

Keywords

• Proteaz, Üretim, Optimizasyon, Bacillus cereus.

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