Directing Biosynthesis

Abstract

The increasingly rapid accumulation of genomic information is revolutionizing natural products discovery. However, the translation of sequence data to chemical products remains a challenge. Here, we detail methods used to circumvent the supply problem of cyanobactin natural products, both by engineered synthesis in E. coli and by using purified enzymes in vitro. Such methodologies exploit nature’s strategies of combinatorial chemistry in the cyanobactin class of RiPP natural products. As a result, it is possible to synthesize a wide variety of natural and unnatural compounds.

New developments in RiPP discovery, enzymology and engineering

“…325 This diversity generating strategy in RiPP biosynthesis complements the alternative approach of hypervariability of core peptide sequences. 304,[326][327][328] Both types of pathways result in high molecular diversity at the cost of catalytic efficiency. 78,325 This section discusses new tailoring reactions reported since 2013, the molecular mechanisms by which they take place, and highlights differences and commonalities in the various strategies used towards natural diversication of RiPPs.…”

Characterization of the Fast and Promiscuous Macrocyclase from Plant PCY1 Enables the Use of Simple Substrates

“…Cyanobactins possess a wide range of desirable and valuable bioactivities including anticancer (cytostatic and cytotoxic), antifouling, immunomodulating, and antineoplastic . An understanding of the cyanobactin biosynthetic machinery has underpinned the production of a wide range of novel cyanobactins in vivo , and in vitro. − While in vivo approachessometimes termed “cell factory”hold the advantage of simplicity, in vitro technologies have the unique capacity of harnessing the power and diversity of chemical synthesis.…”

Biocatalytic synthesis of peptidic natural products and related analogues

“…The first enzyme for the macrocyclization of RiPP peptide backbone was discovered in the biosynthesis of cyanobactins, specifically PatG in the patellamide pathway ( Koehnke et al., 2012 ; McIntosh et al., 2010 ). Cyanobactins are cyclic 6-to-8-amino acid peptides potentially containing multiple modifications, including heterocyclization, oxidation, and prenylation, which are catalyzed by heterocyclases (e.g., PatD), oxidases (e.g., ThcOx), N -terminal proteases (e.g., PatA), C -terminal proteases/cyclases (e.g., PatG), and prenyltransferases (e.g., PatF) ( Sardar et al., 2016 ). Natural cyanobactin analogues display promising biological activities ( Martins and Vasconcelos, 2015 ), e.g., cytotoxicity toward multiple-drug resistant cancers and protease inhibition.…”