caa a22 4500 605498075 CHVBK 20210128100544.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-5880-7 doi (NATIONALLICENCE)springer-10.1007/s00253-014-5880-7 Enzymatic dynamic kinetic resolution of racemic N -formyl- and N -carbamoyl-amino acids using immobilized l - N -carbamoylase and N -succinyl-amino acid racemase [Elektronische Daten] [Pablo Soriano-Maldonado, Francisco Las Heras-Vazquez, Josefa Clemente-Jimenez, Felipe Rodriguez-Vico, Sergio Martínez-Rodríguez] Taking advantage of the catalytic promiscuity of l-carbamoylase from Geobacillus stearothermophilus CECT43 (BsLcar) and N-succinyl-amino acid racemase from Geobacillus kaustophilus CECT4264 (GkNSAAR), we have evaluated the production of different optically pure l-α-amino acids starting from different racemic N-formyl- and N-carbamoyl-amino acids using a dynamic kinetic resolution approach. The enzymes were immobilized on two different solid supports, resulting in improved stability of the enzymes in terms of thermostability and storage when compared to the enzymes in solution. The bienzymatic system retained up to 80% conversion efficiency after 20weeks at 4°C and up to 90% after 1week at 45°C. The immobilization process also resulted in a great enhancement of the activity of BsLcar toward N-formyl-tryptophan, showing for the first time that substrate specificity of l-carbamoylases can be influenced by this approach. The system was effective for the biosynthesis of natural and unnatural l-amino acids (enantiomeric excess (e.e.) >99.5%), such as l-methionine, l-alanine, l-tryptophan, l-homophenylalanine, l-aminobutyric acid, and l-norleucine, with a higher performance toward N-formyl-α-amino acid substrates. Biocatalyst reuse was studied, and after 10 reaction cycles, over 75% activity remained. Springer-Verlag Berlin Heidelberg, 2014 l - N -Carbamoylase nationallicence N -Succinyl-amino acid racemase nationallicence N -Acetyl-amino acid racemase nationallicence l -Norleucine nationallicence l -Homophenylalanine nationallicence l -2-Aminobutyric acid nationallicence l -Tryptophan nationallicence Soriano-Maldonado Pablo Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut Las Heras-Vazquez Francisco Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut Clemente-Jimenez Josefa Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut Rodriguez-Vico Felipe Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut Martínez-Rodríguez Sergio Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 283-291 0175-7598 99:1<283 2015 99 253 https://doi.org/10.1007/s00253-014-5880-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-5880-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Soriano-Maldonado Pablo Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut NATIONALLICENCE 700 1- Las Heras-Vazquez Francisco Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut NATIONALLICENCE 700 1- Clemente-Jimenez Josefa Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut NATIONALLICENCE 700 1- Rodriguez-Vico Felipe Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut NATIONALLICENCE 700 1- Martínez-Rodríguez Sergio Departamento de Química y Física, Universidad de Almería, Campus de Excelencia Internacional Agroalimentario, CEIA3, Edificio CITE I, Carretera de Sacramento s/n., 04120 La Cañada de San Urbano, Almería, Spain aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 283-291 0175-7598 99:1<283 2015 99 253