Enzymatic synthesis ofN‐acyl‐l‐amino acids in a glycerol‐water system using acylase I from pig kidney

Abstract

N-Medium-and long-chain acyl-L-amino acids were enzymatically synthesized from the corresponding L-amino acids and fatty acids using a reverse hydrolysis. Enzymes that are suitable for the synthetic reaction of N-acyl-L-amino acids were screened on the basis of hydrolytic activity toward N-lauroyl-L-glutamic acid as an indicator. Acylase I from pig kidney (EC 3.5.1.14) showed the highest N-acyl-L-amino acid hydrolytic activity among 57 commercially available enzymes tested. Acylase I effectively catalyzed the synthesis of N-lauroyl-L-amino acids except for Nlauroyl-L-proline and N-lauroyl-L-tyrosine in a glycerol-water system. Under the optimized reaction conditions, N-lauroyl-L-arginine and N-lauroyl-L-glutamic acid were obtained in conversions of 82 and 44%, respectively. The equilibrium constants calculated from the conversion obtained were 5. 6, 15.4, 18.0, and 39.4 for the syntheses of N-lauroyl-L-glutamic acid, N α -lauroyl-L-lysine, Nlauroyl-L-glutamine, and N-lauroyl-L-methionine, respectively. NAcyl-L-arginines with myristic acid and palmitic acid as the fatty acid were also synthesized using acylase I.

Amino Acids, Lactic Acid and Ascorbic Acid as Raw Materials for Biocompatible Surfactants

“…The main advantages associated with the use of biocatalysts are mild reaction conditions and high enzymatic specificity, which often eliminate the need for regioselective protection of multifunctional starting materials. This point has been clearly demonstrated by a number of recent reports dealing with the application of enzymes to synthesis and/or modification of amino acids, sugar fatty acid esters, phospholipids and alkyl glycosides [40][41][42][43][44].…”

Increasing the synthesis/hydrolysis ratio of aminoacylase 1 by site-directed mutagenesis

“…Acy1 from porcine kidney has been considered as a biocatalyst for the aqueous one-step synthesis of short-chain and surface-active amphipathic N-acyl-L-amino acids [14,[17][18][19]. Our simulation of the pAcy1-catalyzed amino acid N-acylation reaction based on the kinetic model by Biselli (1992) (Supplementary material, Equation S1 and Fig.…”

“…The advantage of an aminoacylase-catalyzed process consists of the possibility to obtain the desired N-acyl-L-amino acid directly in a single step from the corresponding free L-amino acid and acyl donor, respectively. Strategies to facilitate this reaction for certain amino acid and acyl donor combinations include the use of water miscible solvents in combination with an excess of one substrate and continuous precipitation of the product [12][13][14]. Aminoacylase-1 from pig kidney (pAcy1) displays a marked preference for short-chain acyl moieties and non-branched aliphatic L-amino acids [15,16] and has gained considerable attention as chiral catalyst.…”

“…A number of studies have reported on the use of pAcy1 in reverse hydrolysis, e.g., in the synthesis of short-chain and surface-active amphipathic N-acyl-L-amino acids. The reaction, however, has an unfavorable equilibrium in water and proceeds at a low catalytic rate [14,[17][18][19].S y n t h e s i so f N-acetyl-L-methionine (A-L-Met) by pAcy1 was shown to be most efficient at pH 6 while hydrolysis prevails at higher pH [17,20].Y e t , a decreased concentration of the nucleophilic free amine and thus reduced overall synthetic rates, as well as low enzyme stability precluded robust processes at the acidic pH [21]. pAcy1 belongs to the M20 family of the MH clan of co-catalytic metallopeptidases [22].…”

Eco-Technologies for Obtaining Aminoacid Based Surfactants

“…The most important class of amino acid based surfactants is represented by N-acyl derivatives (acyl amino acids), comprising anionic, cationic, amphoteric or non-ionic surfactants, depending on the ionic nature of the amino acid. Synthesis methods include enzymatic [6][7][8] and chemo-enzymatic [9] processes, but mainly chemical methods due to relatively low production costs [10][11][12]. In this work we have focused on synthesis of surfactants based on hydroxyproline and glygylglycine amino acids, through processes with low environmental impact.…”