Patent ID: 8293511

Claim:
An isolated non-ribosomal peptide synthetase (NRPS) that catalyses formation of the tripeptide hydroxyphenylglycyl-cysteinyl-valine or the tripeptide phenylglycyl-cysteinyl-valine from hydroxyphenylglycine, cysteine and valine or from phenylglycine, cysteine and valine, respectively, the NRPS comprising (i) a first module M1 specific for hydroxyphenylglycine or phenylglycine comprising SEQ ID NO: 2 or SEQ ID NO: 4 or, where the first module M1 is specific for hydroxyphenylglycine the first module M1 comprises a sequence additionally selected from the sixth module of a Calcium-Dependent Antibiotic (CDA) synthetase of a Streptomyces coelicolor or the fourth module of a Chloroerenomycin Synthetase of a Amycolatopsis orientalis or the fifth module of a Chloroerenomycin Synthetase of a Amycolatopsis orientalis, or the seventh module of a Complestatin Synthetase of a Streptomyces lavendulae, and where the first module M1 is specific for phenylglycine the first module M1 comprises a sequence additionally selected from a C-terminal module of a SnbD protein of a Pristinamycin Synthetase of a Streptomyces pristinaspirali, (ii) a second module M2 specific for cysteine comprises SEQ ID NO: 6 or SEQ ID NO: 8 or a second module of an δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase of a Nocardia lactamdurans or a second module of an δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase of a Penicillium chrysogenum or a second module of an δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase of an Acremonium chrysogenum or a second module of an δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase of an Aspergillus nidulans or a domain of the second module of Bacillus subtilis RB14 Iturin Synthetase Protein ItuC, or an amino acid sequence of Penicllium chrysogenum enabling incorporation of the amino acid L-cysteine while being coupled to the amino acid Hpq or Pq, (iii) a third module M3 specific for valine having SEQ ID NO: 10 or a third module of an δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase of a Nocardia lactamdurans or a third module of an δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase of a Penicillium chrysogenum or a third module of an δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase of an Acremonium chrysogenum or a third module of an δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase of an Aspergillus nidulans , the third module M3 enabling incorporation of the amino acid L-valine to hydroxyphenylglycyl-Cysteine or phenylglycyl-Cysteine.