Materials and methods
2.1. Organism and growth conditions
Bacillus spp., potent protease (SBP-114) and lipase (RK-2) producers
were isolated from the soil by enrichment and selective
screening on skim milk agar plate and tributyrin agar plate
respectively. These strains were identified by MIDILABS (New York,
DE, USA) as Bacillus licheniformis and B. subtilus based on 500-bp
analysis and on the partial 16S rRNA gene alignment with the Gene
Bank database (ncbi, 2012) (). The organisms were cultivated at
37 1 C in a bacteriological incubator for 24 h and subsequently
maintained at 4 C in a B.O.D incubator (Yorko, Deluxe-10) by
routine transfers on nutrient agar slants at pH value of 7.0.