Acyl amino acids

摘要

Engineered polypeptides useful in synthesizing acyl amino acids are provided. Also provided are methods of making acyl amino acids using engineered polypeptides. In certain embodiments, an acyl amino acid produced using compositions and/or methods of the present invention comprises cocoyl glutamate.

主权项

1. A method of producing an acyl amino acid, comprising steps of:
providing an engineered polypeptide comprising each of:
a fatty acid linkage domain that is at least 70% identical at the amino acid level to that found in Bacillus subtilis's surfactin synthetase SrfA protein;a peptide synthetase domain that is at least 70% identical at the amino acid level to Bacillus subtilis's surfactin synthetase complex SrfA-A polypeptide subunit's first peptide synthetase domain; anda thioesterase domain that is at least 70% identical at the amino acid level to that found in Bacillus subtilis's surfactin synthetase complex's SrfC subunit;which fatty acid linkage domain, peptide synthetase domain and thioesterase domain are covalently linked; providing fatty acid recognized by the fatty acid linkage domain; providing an amino acid recognized by the peptide synthetase domain; incubating the engineered polypeptide, fatty acid, and amino acid under conditions and for a time sufficient for the fatty acid linkage domain to link the fatty acid to the amino acid to generate an acyl amino acid; incubating the engineered polypeptide and the acyl amino acid under conditions and for a time sufficient for the thioesterase domain to catalyze release of the acyl amino acid from the engineered polypeptide.