Document Detail

Evaluation of substrate promiscuity of an L-carbamoyl amino acid amidohydrolase from Geobacillus stearothermophilus CECT43.
MedLine Citation:
PMID:  20730754     Owner:  NLM     Status:  MEDLINE    
N-carbamoyl-amino-acid amidohydrolase (also known as N-carbamoylase) is the stereospecific enzyme responsible for the chirality of the D- or L-amino acid obtained in the "Hydantoinase Process." This process is based on the dynamic kinetic resolution of D,L-5-monosubstituted hydantoins. In this work, we have demonstrated the capability of a recombinant L-N-carbamoylase from the thermophilic bacterium Geobacillus stearothermophilus CECT43 (BsLcar) to hydrolyze N-acetyl and N-formyl-L-amino acids as well as the known N-carbamoyl-L-amino acids, thus proving its substrate promiscuity. BsLcar showed faster hydrolysis for N-formyl-L-amino acids than for N-carbamoyl and N-acetyl-L-derivatives, with a catalytic efficiency (k(cat)/K(m)) of 8.58 x 10(5), 1.83 x 10(4), and 1.78 x 10(3) (s(-1) M(-1)), respectively, for the three precursors of L-methionine. Optimum reaction conditions for BsLcar, using the three N-substituted-L-methionine substrates, were 65 degrees C and pH 7.5. In all three cases, the metal ions Co(2+), Mn(2+), and Ni(2+) greatly enhanced BsLcar activity, whereas metal-chelating agents inhibited it, showing that BsLcar is a metalloenzyme. The Co(2+)-dependent activity profile of the enzyme showed no detectable inhibition at high metal ion concentrations.
Joaquín Pozo-Dengra; Ana Isabel Martínez-Gómez; Sergio Martínez-Rodríguez; Josefa María Clemente-Jiménez; Felipe Rodríguez-Vico; Francisco Javier Las Heras-Vázquez
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology progress     Volume:  26     ISSN:  1520-6033     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    2010 Jul-Aug
Date Detail:
Created Date:  2010-08-23     Completed Date:  2010-12-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  954-9     Citation Subset:  IM    
Copyright Information:
(c) 2010 American Institute of Chemical Engineers
Dept. de Química-Física, Bioquímica y Química Inorgánica. Edificio C.I.T.E. I., Universidad de Almería, La Cañada de San Urbano, Almería, Spain.
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MeSH Terms
Amidohydrolases / genetics,  metabolism*
Geobacillus stearothermophilus / enzymology*,  metabolism
Hydrogen-Ion Concentration
Substrate Specificity
Reg. No./Substance:
EC 3.5.-/Amidohydrolases; EC 3.5.1.-/N-carbamoylamino acid amidohydrolase

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