RT Journal Article
SR Electronic
T1 Engineering DNA templated nonribosomal peptide synthesis
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.07.27.223297
DO 10.1101/2020.07.27.223297
A1 Hsin-Mei Huang
A1 Philipp Stephan
A1 Hajo Kries
YR 2020
UL http://biorxiv.org/content/early/2020/07/27/2020.07.27.223297.abstract
AB Nanocontainers or macromolecular scaffolds for artificial biocatalytic cascades facilitate sequential enzyme reactions but diffusive escape of intermediates limits rate enhancement. Nonribosomal peptide synthetases (NRPS) naturally form gigantic assembly lines and prevent escape by covalently tethering intermediates. Here, we have built DNA-templated NRPS (DT-NRPS) by adding zinc finger tags to split NRPS modules. The zinc fingers direct the NRPS modules to 9-bp binding sites on a DNA strand, where they form a catalytically active enzyme cascade. DT-NRPS outperform previously reported DNA templated enzyme cascades in terms of DNA acceleration which demonstrates that covalent intermediate channeling is possible along the DNA template. Attachment of assembly line enzymes to a DNA scaffold is a promising catalytic strategy for the sequence-controlled biosynthesis of nonribosomal peptides and other polymers.Competing Interest StatementThe authors have declared no competing interest.