TY  - JOUR
A1  - Soeriyadi, Angela H.
A1  - Ongley, Sarah E.
A1  - Kehr, Jan-Christoph
A1  - Pickford, Russel
A1  - Dittmann, Elke
A1  - Neilan, Brett A.
T1  - Tailoring enzyme stringency masks the multispecificity of a lyngbyatoxin (indolactam alkaloid) nonribosomal peptide synthetase
JF  - ChemBioChem
N2  - Indolactam alkaloids are activators of protein kinase C (PKC) and are of pharmacological interest for the treatment of pathologies involving PKC dysregulation. The marine cyanobacterial nonribosomal peptide synthetase (NRPS) pathway for lyngbyatoxin biosynthesis, which we previously expressed in E. coli, was studied for its amenability towards the biosynthesis of indolactam variants. Modification of culture conditions for our E. coli heterologous expression host and analysis of pathway products suggested the native lyngbyatoxin pathway NRPS does possess a degree of relaxed specificity. Site-directed mutagenesis of two positions within the adenylation domain (A-domain) substrate-binding pocket was performed, resulting in an alteration of substrate preference between valine, isoleucine, and leucine. We observed relative congruence of in vitro substrate activation by the LtxA NRPS to in vivo product formation. While there was a preference for isoleucine over leucine, the substitution of alternative tailoring domains may unveil the true in vivo effects of the mutations introduced herein.
KW  - a domain
KW  - indolactams
KW  - MbtH
KW  - natural products
KW  - teleocidin
Y1  - 2021
U6  - https://doi.org/10.1002/cbic.202100574
SN  - 1439-4227
SN  - 1439-7633
VL  - 23
IS  - 3
PB  - Wiley-VCH
CY  - Weinheim
ER  -