A Chemo-Enzymatic Road Map to the Synthesis of CoA Esters

Abstract

Coenzyme A (CoA) is a ubiquitous cofactor present in every known organism. The thioesters of CoA are core intermediates in many metabolic processes, such as the citric acid cycle, fatty acid biosynthesis and secondary metabolism, including polyketide biosynthesis. Synthesis of CoA-thioesters is vital for the study of CoA-dependent enzymes and pathways, but also as standards for metabolomics studies. In this work we systematically tested five chemo-enzymatic methods for the synthesis of the three most abundant acyl-CoA thioester classes in biology; saturated acyl-CoAs, α,β-unsaturated acyl-CoAs (i.e., enoyl-CoA derivatives), and α-carboxylated acyl-CoAs (i.e., malonyl-CoA derivatives). Additionally we report on the substrate promiscuity of three newly described acyl-CoA dehydrogenases that allow the simple conversion of acyl-CoAs into enoyl-CoAs. With these five methods, we synthesized 26 different CoA-thioesters with a yield of 40% or higher. The CoA esters produced range from short-to long-chain, include branched and α,β-unsaturated representatives as well as other functional groups. Based on our results we provide a general guideline to the optimal synthesis method of a given CoA-thioester in respect to its functional group(s) and the commercial availability of the precursor molecule. The proposed synthetic routes can be performed in small scale and do not require special chemical equipment, making them convenient also for biological laboratories.

Characterization of an Orphan Type III Polyketide Synthase Conserved in Uncultivated “Entotheonella” Sponge Symbionts

“…Reactions were performed in a total volume of 200 μL and were incubated for 3 h at 37 °C. Starter units were kindly provided by the group of Erb . Reactions for time‐course experiments and isolation of polyketides contained phosphate buffer (500 m m , pH 6) with malonyl‐CoA (300 μ m ) and palmitoyl‐CoA (100 μ m ).…”

Enzymatic Halogenation and Dehalogenation Reactions: Pervasive and Mechanistically Diverse

“…44 Clearly, a co-crystal structure of an acyl- S -CP FDH in complex with its physiological substrate will go a long way in verifying or refuting the above hypotheses. Advances in the biosynthesis of acyl-coenzyme A thioesters, 122,123 mass spectrometry based analytical techniques 124 that circumvent alkaline offloading of substrates and products from acyl- S -CPs prior to the characterization, and synthesis of molecular probes to interrogate flavin-dependent enzymes 125 will undoubtedly aid in answering these mechanistic conundrums that have persisted for more than a decade.…”

Anaerobic degradation of phenanthrene by a sulfate‐reducing enrichment culture

“…A chemical standard of 2-phenanthroyl-CoA or 4-and 9-phenanthroic acid was synthesized in a small-scale reaction following a two-step procedure (Kawaguchi et al, 1981;Peter et al, 2016). First, 2-phenanthroic acid was activated with carbonyldiimidazole (CDI) in tetrahydrofuran (THF).…”