Heterologous Expression of a Putative ClpC Chaperone Gene Leads to Induction of a Host Metabolite

Abstract

Genome mining provides exciting opportunities for the discovery of natural products. However, in contrast to traditional bioassay-guided approaches, challenges of genome mining include poor or no expression of biosynthetic gene clusters (BGCs). Additionally, given that thousands of BGCs are now available through extensive genome sequencing, how does one select BGCs for discovery? Synthetic biology techniques can be used for BGC refactoring and activation, whereas resistancegene-directed genome mining is a promising approach to discover bioactive natural products. Here we report the selection of a BGC by applying a resistance-gene-directed approach, cloning of the silent BGC from Micromonospora sp. B006, promoter exchange, and heterologous expression in Streptomyces coelicolor M1152. While we have yet to identify the encoded compound, we unexpectedly observed induction of a host metabolite, which we hypothesize is due to the presence of a caseinolytic protease C (ClpC) chaperone gene in the BGC, suggesting that ClpC chaperones may be used for BGC activation.

Chemoselectivity Strategy Based on B-Label Integrated with Tailored COF for Targeted Metabolomic Analysis of Short-Chain Fatty Acids by UHPLC-MS/MS

“…The chemoselective extraction of targeted metabolites is an emerging research topic, a powerful means for the studying of systemic metabolic profiling, which provides strong specificity, high detection sensitivity, and accurate quantification for the analysis of metabolites. , Short-chain fatty acids (SCFAs), straight or branched chain aliphatic organic acids with 2 to 6 carbon atoms (such as acetic acid, propanoic acid, isobutyric acid, and propanedioic acid), − are conducive to regulate the metabolism of glucose, cholesterol, and lipids in peripheral tissues − and play key roles in regulating appetite , and inflammation and maintaining intestinal and immune function. , SCFAs involve many pivotal metabolic processes, such as amino acid metabolism, fatty acid metabolism, and energy metabolism, among which short-chain binary fatty acids, such as α-ketoglutaric acid, succinic acid, and fumaric acid, are also the main intermediate products of the tricarboxylic acid (TCA) cycle. , Many literature reports have suggested that these SCFAs serve as potential biomarkers for disease diagnosis, evaluation of new drugs, and elucidation of drug action mechanisms. − …”