RESUMEN

About 110 newly isolated halophilic and halotolerant bacteria were screened for protease production. A moderately halophilic strain (CJ4), isolated from Chott Eldjerid Hypersaline lake in Tunisia, showed the highest activity on agar plate and was then selected. The biochemical and physiological characterization of the isolate along with the 16S rRNA sequence analysis placed it in the genus Halobacillus. Protease production was maximal at 120 g/L NaCl (2 M) and it started from the post-exponential phase reaching a maximum level at the early decline phase of bacterial growth. Protease activity was optimal at 0.4 M NaCl, pH 9 and 45 °C. It showed an excellent stability over wide ranges of temperatures (30-60 °C), NaCl concentrations (0-5 M), and pH values (5-10), which make it a good candidate for industrial applications at harsh conditions. Crude protease was strongly inhibited by PMSF revealing the dominance of serine proteases. Protease activity exhibited high stability in the presence of several organic solvents and detergent additives. These findings make Halobacillus sp. CJ4 protease with a great interest for many biotechnological applications at high salt or low water content such as peptide synthesis and detergent formulation.

Asunto(s)

Halobacillus/enzimología , Serina Proteasas/aislamiento & purificación , Serina Proteasas/metabolismo , Técnicas de Tipificación Bacteriana , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Detergentes/metabolismo , Inhibidores Enzimáticos/metabolismo , Estabilidad de Enzimas , Halobacillus/clasificación , Halobacillus/genética , Halobacillus/fisiología , Concentración de Iones de Hidrógeno , Lagos/microbiología , Fluoruro de Fenilmetilsulfonilo/metabolismo , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Serina Proteasas/química , Cloruro de Sodio/metabolismo , Solventes/metabolismo , Temperatura , Túnez