Production of enantiomerically pure D-Phenylglycine using Pseudomonas aeruginosa 10145 as biocatalyst

Abstract

-Different bacterial strains were screened to detect nitrilase and/or nitrile hidratase/amidase activities towards benzonitrile, to be used as biocatalyst to produce enantiomerically pure non-proteinogenic amino acids using amino nitriles as starting material. The best biocatalyst found was Pseudomonas aeruginosa 10145, which showed high enzyme activities. Whole cells were used as catalyst for the transformation of 2-phenyl-2-amino-acetonitrile for the corresponding D-phenylglycine. The percentage conversion was followed by chiral HPLC. After 1 hour reaction 18% of 2-phenyl-2-amino-acetonitrile was converted into D-phenylglycine with an enantiomeric excess of over 95%. When an inducer was added to the media, an increase in nitrile hydrolyzing activities was detected, hence leading to total conversion of (R)-2-phenyl-2-amino-acetonitrile to the corresponding amino acid in 30 min reaction. The isolated yield of the target product was 50% and its characterization was performed by polarimetry, chiral HPLC, IR-FT spectroscopy and GC-MS.

Nitrilases in nitrile biocatalysis: recent progress and forthcoming research

“…NDB 1165 nitrilase was induced by propionitrile [49]. Benzonitrile was used as the inducer of nitrilase from P. aeruginosa 10145, in which cell growth was inhibited by the addition of benzonitrile at the beginning and thus the inducer was added after cultivation for 24 h [51]. The effects of different inducers on substrate specificity of A. faecalis MTCC 10757 were investigated by Nageshwar et al, and acrylonitrile was the most preferred inducer and substrate [61].…”

Enantioresolution of Amino Acids: A Decade’s Perspective, Prospects and Challenges

“…Most of the basic methods and principles reported in literature for enantiomeric resolutions are based on experimental investigations carried out on amino acids or through their application as chiral selectors. Demand for (single) enantiomerically pure AAs has continued to grow in such areas as pharmaceuticals [4], biochemistry [5], archeology [6], chiral synthesis [7], etc. The syntheses of optically active compounds (whether of pharmaceutical or synthetic importance, or as promising candidates as chiral ligands and auxiliaries in asymmetric syntheses) result in the formation of a mixture of products with one enantiomer predominating.…”

ENANTIOPURE R(-)-3-AMINOISOBUTYRIC ACID SYNTHESIS USING Pseudomonas aeruginosa AS ENANTIOSPECIFIC BIOCATALYST

“…Chiral HPLC was performed by the protocol described by Alonso et al (2008), slightly modified. It was conducted at room temperature with a Nucleosil Chiral-1 column (4.6 x 250 mm, Macherey-Nagel) using 1mM copper sulfate as the mobile phase at a flow rate of 0.7 mL.min -1 .…”