Enzyme Catalyzed Formation of CoA Adducts of Fluorinated Hexanoic Acid Analogues using a Long-Chain acyl-CoA Synthetase from Gordonia sp. Strain NB4-1Y

• Published in: Biochemistry (Easton) 2024-09, Vol.63 (17), p.2153-2165
• Main Authors: Mothersole, Robert G., Mothersole, Mina K., Goddard, Hannah G., Liu, Jinxia, Van Hamme, Jonathan D.
• Format: Article
• Language: English
• Subjects: Acyl Coenzyme A - chemistry; Acyl Coenzyme A - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Caproates - chemistry; Caproates - metabolism; Coenzyme A - chemistry; Coenzyme A - metabolism; Coenzyme A Ligases - chemistry; Coenzyme A Ligases - genetics; Coenzyme A Ligases - metabolism; enzyme kinetics; family; Gordonia Bacterium - enzymology; Gordonia Bacterium - genetics; Gordonia Bacterium - metabolism; Halogenation; hexanoic acid; Kinetics; long-chain-fatty-acid-CoA ligase; metabolites; propionic acid; soil bacteria; Substrate Specificity
• ISSN: 0006-2960; 1520-4995; 1520-4995
• DOI: 10.1021/acs.biochem.4c00336

Per and polyfluoroalkyl substances (PFAS) are a large family of anthropogenic fluorinated chemicals of increasing environmental concern. Over recent years, numerous microbial communities have been found to be capable of metabolizing some polyfluoroalkyl substances, generating a range of low-molecular-weight PFAS metabolites. One proposed pathway for the microbial breakdown of fluorinated carboxylates includes β-oxidation, this pathway is initiated by the formation of a CoA adduct. However, until recently no PFAS-CoA adducts had been reported. In a previous study, we were able to use a bacterial medium-chain acyl-CoA synthetase (mACS) to form CoA adducts of fluorinated adducts of propanoic acid and pentanoic acid but were not able to detect any products of fluorinated hexanoic acid analogues. Herein, we expressed and purified a long-chain acyl-CoA synthetase (lACS) and a A461K variant of mACS from the soil bacterium Gordonia sp. strain NB4–1Y and performed an analysis of substrate scope and enzyme kinetics using fluorinated and nonfluorinated carboxylates. We determined that lACS can catalyze the formation of CoA adducts of 1:5 fluorotelomer carboxylic acid (FTCA), 2:4 FTCA and 3:3 FTCA, albeit with generally low turnover rates (