Enhanced Production of Extra Cellular Alkaline Protease in Bacillus Circulance Through Plasmid Transfer

Five local strains of Bacillus spp. were screened for alkaline protease production. They were Bacillus circulance, Bacillus cereus, Bacillus alvei, Bacillus sphaericus and Bacillus pumilus .Bacillus alvei revealed the highest alkaline protease production, while Bacillus circulance had the lowest productivity among all strains. All strains were characterized on the basis of resistance to either ampicillin (Ap) or tetracycline (Tc) antibiotics as a genetic marker. Bacillus strains were resistant to both antibiotics, (Ap) and (Tc), except Bacillus circulance which was sensitive to both. Due to their antibiotic resistent patterns, all strains were used as sources of plasmids (donor strains) to transform Bacillus circulance to study their effect in alkaline protease productivity. Eight Bacillus circulance transformants were isolated and selected according to their antibiotic resistance. Variation in alkaline protease production levels was detected in all transformants, i.e., 14.7 to 330 U/ml. Up to seven and half times more alkaline protease activity were found in transformants comparing with the recipient strain production .The three highest transformants in enzyme production were BC.NRC4, BC.NRC3 and BC.NRC5,i.e.,330,305 and 227.5 U/ml,respectively . Plasmid profile studies of recipient strain, Bacillus alvei, Bacillus sphaericus (donor strains) and the three highest transformants were done. Results confermed that transfer of plasmids to these transformants. The promising genetically improved strains produced in this study could be used commercially and the system described here could be used also in higher alkaline protease production of other Bacillus strains.

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