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  • Title: A genetic system yields self-cleaving inteins for bioseparations.
    Author: Wood DW, Wu W, Belfort G, Derbyshire V, Belfort M.
    Journal: Nat Biotechnol; 1999 Sep; 17(9):889-92. PubMed ID: 10471931.
    Abstract:
    A self-cleaving element for use in bioseparations has been derived from a naturally occurring, 43 kDa protein splicing element (intein) through a combination of protein engineering and random mutagenesis. A mini-intein (18 kDa) previously engineered for reduced size had compromised activity and was therefore subjected to random mutagenesis and genetic selection. In one selection a mini-intein was isolated with restored splicing activity, while in another, a mutant was isolated with enhanced, pH-sensitive C-terminal cleavage activity. The enhanced-cleavage mutant has utility in affinity fusion-based protein purification. These mutants also provide new insights into the structural and functional roles of some conserved residues in protein splicing.
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