[Design of hybrid metalloproteinases from Bacillus]

Possibility of using recombination mechanisms for the construction of hybrids between relative proteins containing highly homologous regions has been demonstrated. In order to design hybrid neutral proteases (NPr) NPr B. amyloliquefaciens--NPr B. brevis genes encoding these enzymes have been cloned into the same plasmid in tandem orientation with subsequent recombination between them. With the help of sequential inactivation we have managed to ensure efficient selection at intermediate stages as well as at the completion stage of construction when the desirable hybrid forms exhibited proteolytic activity. Constructions containing expressed genes of the hybrid neutral proteases NPr B. amyloliquefaciens--NPr B. brevis were obtained. The presence of several regions of high homology between the genes of the two Bacilli neutral proteinases determines the possibility of obtaining of various variants of hybrid proteins with different properties.