Coenzyme A ligases play an important role in metabolism by catalyzing the activation of carboxylic acids. In this study we describe the synthesis of aminoacyl-coenzyme As (CoAs) catalyzed by a CoA ligase from Penicillium chrysogenum. The enzyme accepted medium-chain length fatty acids as the best substrates, but the proteinogenic amino acids L-phenylalanine and L-tyrosine, as well as the non-proteinogenic amino acids D-phenylalanine, D-tyrosine and (R)- and (S)-beta-phenylalanine were also accepted. Of these amino acids, the highest activity was found for (R)-beta-phenylalanine, forming (R)-beta-phenylalanyl-CoA. Homology modeling suggested that alanine 312 is part of the active site cavity, and mutagenesis (A312G) yielded a variant that has an enhanced catalytic efficiency with beta-phenylalanines and D-alpha-phenylalanine. (C) 2011 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.
|Nummer van het tijdschrift||6|
|Status||Published - 23-mrt.-2011|