Leader sequence inducing a post-translational modification of po

Details Leader sequence inducing a post-translational modification of po Leader sequence inducing a post-translational modification of po

1995-06-05

1999-03-23

Degen, Nancy

Chemistry: molecular biology and microbiology

Treatment of micro-organisms or enzymes with electrical or...

Modification of viruses

435 691, 4352523, 4353201, 530300, 530324, 536 231, C12N 1511, C12P 2102, C07H 2104, C07K 1932

Patent

active

058858113

ABSTRACT:
The method by which polypeptides having residues other than the 20 common amino acids are made is established. A leader peptide sequence (SEQ ID NO: 7) 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
Wei Lui & J. Norman Hanse, "Enhancement of the Chemical and Antimicrobial properties of Subtiline by Site-directed Mutagenesis", The Journal of Biological Chemistry, (1992) The American Society for Biochemistry and Molecular Biology, Inc., vol., 267, No. 35, Dec. 15, 1992, pp. 25078-25085 .

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