Purification and characterization of an extracellular proteinase from Brevibacterium linens ATCC 9174

Abstract

An extracellular serine proteinase from Brevibacterium linens ATCC 9174 was purified to homogeneity, pH and temperature optima were 8.5 and 50°C, respectively. The results for the molecular mass of the proteinase were 56 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 126 kDa by gel filtration, indicating that the native enzyme exists as a dimer. Mg2+ and Ca2+ activated the proteinase, as did NaCl; however, Hg2+, Fe2+, and Zn2+ caused strong inhibition. The sequence of the first 20 N- terminal amino acids was NH2-Ala-Lys-Asn-Asp-Ala-Val-Gly-Gly-Met-Gly-Tyr- Leu-Ser-Met-Ile-Pro-Ser-Gln-Pro-Gly.