Ribosome-mediated synthesis of natural product-like peptides via cell-free translation

Abstract

Peptide natural products (PNPs) represent a unique class of compounds known for their fascinating structural motifs with important biological activities. Lately, PNPs have garnered a lot of interest for their application in drug discovery. Nevertheless, lack of diversity oriented synthetic/biosynthetic platforms to generate large natural product-like libraries has limited their development as peptide therapeutics. The promiscuity of cell-free translation has allowed for the synthesis of artificial PNPs having complex structural features. Modified cell-free translation systems coupled with the display technologies have generated diverse natural product-like peptide libraries and led to the discovery of several biologically active molecules. Such technologies have drastically decreased the time to obtain peptide drug leads and therefore, revolutionized the field of peptide drug discovery. In this account, we review recent developments in the synthesis of natural product-like peptides via cell-free translation.

Multiplex transcriptional characterizations across diverse bacterial species using cell‐free systems

“…Cell‐free systems, made up of either individually reconstituted cellular components (Shimizu et al , ; Wang et al , ; Villarreal et al , ) or cell lysates (Jewett et al , ; Garamella et al , ), can support a variety of catalytic reactions in vitro when supplied with energy sources, cofactors, and ions (Calhoun & Swartz, , ; Jewett et al , ). These cell‐free approaches have facilitated the development of therapeutically useful natural products (Dudley et al , ; Maini et al , ) and biologics with non‐standard amino acids (Martin et al , ) or chemical moieties otherwise challenging to synthesize (Jaroentomeechai et al , ). They have also been used to generate viable phage particles directly from purified phage DNA (Shin et al , ; Garamella et al , ; Rustad et al , ) and detect pathogens in emerging diagnostic applications (Pardee et al , ; Takahashi et al , ).…”

Rapid and scalable preparation of bacterial lysates for cell-free gene expression

“…Although in this paper we focused on production of lysates for cell-free expression, they could also be employed for other emerging synthetic biology and biotechnology applications, such as cell-free proteomics (4), metabolic engineering (1) or natural product and natural-like product biosynthesis (8)(9)(10). Given the pace of development of cell-free synthetic biology and biotechnology, and the rapidly growing number of researchers working in these fields, we believe that this accessible method of lysate preparation will be of benefit to a large number of researchers.…”

Genetically Encoded Libraries of Constrained Peptides

“…An engineered ribozyme that is flexibly capable of esterifying activated amino acids on full‐length in vitro transcribed tRNAs has been generated . As a result, peptides can be composed of over 400 different constituents instead of the ordinary 20 amino acids.…”