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de Rond, T.

Publications and source records attributed to de Rond, T..

2 recordsLinked to original sources

Discovery and Biosynthesis of Sulfenicin and Its New-to-Nature Acylsulfenic Acid Functional Group

Lifes organic molecules are built with diverse functional groups that enable biology by fine tuning intimate connections through time and space. As such, the discovery of new-to-nature functional groups can expand our understanding of the natural world and motivate new applications in biotechnology and biomedicine. Herein we report the genome-aided discovery of sulfenicin, a novel polyketide-nonribosomal peptide hybrid natural product from a marine Streptomyces bacterium bearing a unique acylsulfenic acid functionality. Through a series of heterologous biosynthesis, functional genetics, and enzymatic reconstitution experiments, we show that this previously described synthetic functional group is biologically assembled by a set of enzymes from both primary and secondary metabolism, including a novel flavin-dependent S-hydroxylase that hydroxylates a thiocarboxylic acids sulfur atom. While the sulfenicin biosynthetic gene cluster is presently without parallel in public databases, acylsulfenic acid-encoding enzymes are widely distributed in bacterial genomes, implying that this labile functional group may similarly have a broad distribution among specialized metabolites.

biochemistry↗

Co-Occurrence of Enzyme Domains Guides the Discovery of an Oxazolone Synthetase

Multidomain enzymes are cellular machines that orchestrate two or more catalytic activities to carry out metabolic transformations with increased control and speed. Our understanding of these enzymes capabilities drives progress in fundamental metabolic research, biocatalysis, and human health. Here, we report the development of a new genome mining approach for the targeted discovery of novel biochemical transformations through the analysis of co-occurring enzyme domains (CO-ED) in a single protein. CO-ED was designed to identify unannotated multifunctional enzymes for functional characterization and discovery based on the premise that linked enzyme domains have evolved to function collaboratively. Guided by CO-ED, we targeted an unannotated predicted ThiF-nitroreductase di-domain enzyme found in more than 50 proteobacteria. Through heterologous expression and biochemical reconstitution, we discovered a series of new natural products containing the rare oxazolone (azlactone) heterocycle and characterized the di-domain enzyme as the first reported oxazolone synthetase in biology. This enzyme has the potential to become a valuable biocatalyst for the production of versatile oxazolone synthetic intermediates. This proof-of-principle experiment validates CO-ED-guided genome mining as a new method with potential broad utility for both the discovery of novel enzymatic transformations and the functional gene annotation of multidomain enzymes. TOC graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=109 SRC="FIGDIR/small/147165v2_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@20cb16org.highwire.dtl.DTLVardef@f54df0org.highwire.dtl.DTLVardef@cffa51org.highwire.dtl.DTLVardef@3b085d_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗