Identification of three extracellular proteases from Bacillus subtilis KCTC 3014

摘要: Three extracellular proteases (Vpr, peptidase T, and subtilisin) were identified from the culture supernatant of Bacillus subtilis KCTC 3014. All proteins partially purified as a mature form by using DEAE-cellulose ion-exchange column chromatography. Their activities determined zymography densitometry. The relative molecular masses Vpr T (PepT) to be 68 48 kDa SDS-PAGE zymography, respectively. However, subtilisin formed "binding mode" at top separating gel. After denaturation boiling 100°C for 5 min, its mass was 29 kDa, whereas activity lost. optimal pH Vpr, PepT, 9.0, 6.0-7.0, 7.0-8.0, temperature 40, 50, 40°C, Inhibitor test revealed that serine PepT metalloprotease. Interestingly, we found showed no enzyme on 2DE zymogram genes, vpr, pepT, apr (encoding protein), cloned their nucleotide deduced amino acid sequences determined.