CAUSING A PLURALITY OF MUTATION IN ARBITRARY POSITION OF PROTEIN CODING REGION

摘要

Methods and vectors are provided for the production of procaryotic carbonyl hydrolases in recombinant systems. DNA which encodes such hydrolases are mutated at predetermined regions by known methods or by a novel point mutagenesis method in order to generate mutant hydrolases. Particular point mutations in carbonyl hydrolases such as subtilisin result in modifications of oxidation stability, Km, Kcat, Kcat/Km ratio substrate specificity, specific activity or pH-activity profiles. These mutated hydrolases are particularly useful in laundry compositions. Mutations in the genes encoding the subtilisin or neutral protease of bacillus yield substantially normally sporulating bacillus strains which are incapable of excreting subtilisin or neutral protease. Such strains are useful in the recombinant synthesis of heterologous proteins.