Exploring the synthetic applicability of a new carboxylic acid reductase from Segniliparus rotundus DSM 44985

Adenylation Activity of Carboxylic Acid Reductases Enables the Synthesis of Amides

“…Amidation would be of particular interest because many natural amide synthetases,s uch as NRPS [14] and ATP-grasp enzymes [15,16] ,s uffer from very narrow substrate specificity,w hich limits their applications in biocatalysis.A lternatively,h ydrolases (such as lipases [17] and proteases [18] ), which are commonly used for amide formation, [2] require systems in which very little water is present, such as organic solvents,todrive the reaction towards amide bond formation. [19][20][21][22] Four CARc andidates (CARmm from Mycobacterium marinum, [3] CARni from Nocardia iowensis, [4] CARtp from Tsukamurella paurometabola, [10] and CARse from Segniliparus rotundus, [23] )were produced recombinantly in Escherichia coli,w ith the coexpressed gene for the Bacillus subtilis phosphopantetheinyl transferase (PPTase;S fp), [24] which is required for post-translational addition of the PPant group. [4] These CARs were purified (Supporting Information, Figure S1) and the amidation of carboxylic acid substrates 1-6 was tested.…”

Consolidated production of coniferol and other high-value aromatic alcohols directly from lignocellulosic biomass

“…Although these CARs had their activities evaluated against diverse acids previously, none of these studies provided a detailed comparison of diverse CARs using ferulic acid as a substrate. 23,[27][28][29] The relative activity of the enzyme candidates for the single reduction of 1 to 2 were, Nocardia Iowensis (NiCAR), 27 Segniliparus rugosus (SrCAR), 25,28 Segniliparus rotuduns (SroCAR), 25 Mycobacterium marinum (mCAR) 25 and Tsukamurella paurometabola (TpCAR) 21 with 100%, 100%, 91%, 80% and 42% respectively (Table 1). Both, NiCAR and SroCAR have been previously demonstrated to reduce 1, and NiCAR was reported to have greater activity against this substrate.…”

“…25 However, the reduction of ferulic acid to coniferol has not been explored using this proposed catalytic pathway, and indeed to our knowledge there are no reports of such a cascade using heterologous AKR enzymes. 23,[27][28][29][30] The generation of these chemicals from abundant biomass sources has many advantages including use of mild reaction conditions, compatibility with aqueous media, sourcing of catalysts from renewable feedstocks and chemo-, regio-and enantioselectivity. In this study we explored the use of both an in vitro enzymatic and whole-cell biocatalytic route to release ferulic acid from biomass sources and convert it directly into coniferol ( Fig.…”

Characterization of Type IV Carboxylate Reductases (CARs) for Whole Cell‐Mediated Preparation of 3‐Hydroxytyrosol

“…[14,[18][19][20] Further representatives of fungal CARs are not only needed to diversify the basis of phylogenetic analyses and corroborate conclusions from such analyses, but also to eventually assemble a repertoire of new enzymes with different characteristics. The protein sequences of NiCAR, [21,22] MmCAR, [8] SroCAR [23] and AtCAR [17,20] were taken as templates for a BLASTp search with the restriction to organisms with experimentally confirmed ability to reduce carboxylic acids. Hit sequences were further analyzed for the presence of essential key residues and motifs.…”

“…A phylogenetic tree of fungal and bacterial CAR enzymes, categorized in four subtypes. Newly discovered sequences and previously described CARs [8,14,15,18,19,23] were incorporated in the phylogenetic analysis. Green branches represent CARs investigated in this study.…”