Subtilisin with increased thermal stability

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase

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4351721, 4351723, 43525231, 435320, 536 27, 25217412, 935 14, 935 29, 935 74, C12N 956, C12N 1500, C12N 120, C07H 1512

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049802887

ABSTRACT:
Cloned DNA is mutated by creating a single-stranded target region in a cloned DNA segment, and introducing a mutation into the single-stranded target region by treating the target region with a chemical or biological mutagenizing agent capable of introducing mutations into single-stranded DNA. The mutated target region then is rendered double-stranded and a microorganism is transformed with the mutated double-stranded DNA present in an expression vector. The transformed microorganism is cultivated under conditions wherein the mutated DNA is expressed to form an expression product, and the expression product is screened to identify a desired mutation in the DNA segment. Mutant subtilisins of enhanced thermal stability are also disclosed.

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