Leader sequence inducing a post-translational modification of po

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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4351723, 4352523, 43525231, 4353201, 536 231, C12P 2102, C07H 2104, C12N 121, C12N 1575

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059142502

ABSTRACT:
The method by which polypeptides having residues other than the 20 common amino acids are made is established. A leader peptide sequence and its gene are identified which induce or assist post-translational modifications of Cys, Thr and Ser in prokaryotes. The leader sequence may be used to induce the presence of covalent bonding sites in polypeptides and can be expressed by either naturally occurring or artificial means. Further, a subtilin mutant substituting isoleucine for Glu.sub.4 of the native sequence exhibits a 57-fold improvement in stability, resisting modification of the dehydroalanine residue at position 5. This stable mutant exhibits 3-4 times the specific activity, in suppression of bacterial spore outgrowth, of the native bacteriocin. A method for site-specific mutagenesis, as well as the resulting mutant gene, plasmid and transformant is similarly set forth.

REFERENCES:
patent: 5218101 (1993-06-01), Hansen
patent: 5516682 (1996-05-01), Hansen
patent: 5576420 (1996-11-01), Hansen

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