Preparation, purification, biochemistry and molecular characterization of a novel protease SAP HM from Bacillus licheniformis k7a strain for detergent and peptide synthesis

摘要

The invention relates to the uniform purification of a new heat stable alkaline protease saphm prepared by Bacillus licheniformis k7a strain and the biochemical and molecular characteristics of the enzyme. The strain was newly isolated from soil samples in Hassi messuad, Algeria. After 24 hours of planting, the optimal yield of Erlenmeyer was 12500 U / ml to 45 ° C. The purification technology used in this workThe invention provides a uniform enzyme solution with a molecular weight of 30 kDa, which is determined by SDS PAGE. Its purity was also verified by sequencing the NH2 terminals of 26 AA (aqtvpqgiplikaekvqagfdgarv).It showed a high degree of homogeneity with serine Bacillus protease, 96% The optimal protease activity was obtained at pH 10 and 70 ° C on casein. The enzyme is basically stable within 24 hours of alkaline pH (6-12). It is a kind of enzyme belonging to the family of serum proteases. In fact, it is inhibited by serum enzyme specific inhibitor (PMSF).Page: 1 The thermal activity and thermal stability of 2 mm Ca and 10% sorbitol were greatly improved. Saphm has a wide range of specificity on protein matrix, and has better catalytic efficiency than alkaline protease with more than 2.5 liters.Surfactants (Tween 20, Tween 80 and Triton X-100) and bleachers (perborate sodium and h102) were used. Compared with liquid and solid detergents (pril-isis, tide, dipex, nadhif, Ariel, dixan and skip), the saphm protease has significant stability and compatibility. In addition, saphm protease was added to the detergent solution to improve the performance of pril Isis detergent to improve the bleaching of blood pollutants. On the other hand, the saphm gene encoding the new saphm protease was cloned,It was sequenced and expressed in E. coli BL21 (DE3). Recombinant protein ("rsaphm") s expressed in extracellular spaceIt has the same characteristics as native protein ("saphm"). In the end, it seems that phsams are well tolerated for the preparation of detergents.