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Literature summary for 3.5.1.87 extracted from

  • Soriano-Maldonado, P.; Rodriguez-Alonso, M.; Hernandez-Cervantes, C.; Rodriguez-Garcia, I.; Clemente-Jimenez, J.; Rodriguez-Vico, F.; Martinez-Rodriguez, S.; Las Heras-Vazquez, F.
    Amidohydrolase process: expanding the use of L-N-carbamoylase/N-succinyl-amino acid racemase tandem for the production of different optically pure L-amino acids (2014), Process Biochem., 49, 1281-1287.
No PubMed abstract available

Application

Application Comment Organism
synthesis development of a bienzymatic biocatalyst system comprising an N-succinylamino acid racemase from Geobacillus kaustophilus CECT4264 and the enantiospecific L-N-carbamoylase from Geobacillus stearothermophilus CECT43. The biocatalyst system is able to produce optically pure natural and non-natural L-amino acids starting from racemic mixtures of N-acetyl-, N-formyl- and N-carbamoyl-amino acids by dynamic kinetic resolution. The fastest conversion rate is found with N-formyl-aminoacids, followed by N-carbamoyl- and N-acetyl-amino acids, and the an N-succinylamino acid racemase proves to be the limiting step of the system due to its lower specific activity, overview Geobacillus stearothermophilus

Cloned(Commentary)

Cloned (Comment) Organism
gene Bslcar, recombinant overexpression in Escherichia coli strain BL21 using the pJAVI80Rha plasmid Geobacillus stearothermophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ essential for activity Geobacillus stearothermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
N-carbamoyl-DL-methionine + H2O Geobacillus stearothermophilus
-
L-methionine + CO2 + NH3
-
?
N-carbamoyl-DL-methionine + H2O Geobacillus stearothermophilus CECT43
-
L-methionine + CO2 + NH3
-
?
N-formyl-DL-methionine + H2O Geobacillus stearothermophilus
-
L-methionine + CO2
-
?
N-formyl-DL-methionine + H2O Geobacillus stearothermophilus CECT43
-
L-methionine + CO2
-
?

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus Q53389 gene Bslcar
-
Geobacillus stearothermophilus CECT43 Q53389 gene Bslcar
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the enzyme is a strict enantiospecific L-N-carbamoylase. Development of a bienzymatic system comprising an N-succinylamino acid racemase from Geobacillus kaustophilus CECT4264 and the enantiospecific L-N-carbamoylase from Geobacillus stearothermophilus CECT43. The biocatalyst system is able to produce optically pure natural and non-natural L-amino acids starting from racemic mixtures of N-acetyl-, N-formyl- and N-carbamoyl-amino acids by dynamic kinetic resolution. The fastest conversion rate is found with N-formyl-aminoacids, followed by N-carbamoyl- and N-acetyl-amino acids, and the an N-succinylamino acid racemase proves to be the limiting stepof the system due to its lower specific activity, overview Geobacillus stearothermophilus ?
-
?
additional information the enzyme is a strict enantiospecific L-N-carbamoylase. Development of a bienzymatic system comprising an N-succinylamino acid racemase from Geobacillus kaustophilus CECT4264 and the enantiospecific L-N-carbamoylase from Geobacillus stearothermophilus CECT43. The biocatalyst system is able to produce optically pure natural and non-natural L-amino acids starting from racemic mixtures of N-acetyl-, N-formyl- and N-carbamoyl-amino acids by dynamic kinetic resolution. The fastest conversion rate is found with N-formyl-aminoacids, followed by N-carbamoyl- and N-acetyl-amino acids, and the an N-succinylamino acid racemase proves to be the limiting stepof the system due to its lower specific activity, overview Geobacillus stearothermophilus CECT43 ?
-
?
N-carbamoyl-DL-2-aminoburyric acid + H2O
-
Geobacillus stearothermophilus L-2-aminobutyric acid + CO2 + NH3
-
?
N-carbamoyl-DL-2-aminoburyric acid + H2O
-
Geobacillus stearothermophilus CECT43 L-2-aminobutyric acid + CO2 + NH3
-
?
N-carbamoyl-DL-homophenylalanine + H2O
-
Geobacillus stearothermophilus L-homophenylalanine + CO2 + NH3
-
?
N-carbamoyl-DL-homophenylalanine + H2O
-
Geobacillus stearothermophilus CECT43 L-homophenylalanine + CO2 + NH3
-
?
N-carbamoyl-DL-methionine + H2O
-
Geobacillus stearothermophilus L-methionine + CO2 + NH3
-
?
N-carbamoyl-DL-methionine + H2O
-
Geobacillus stearothermophilus CECT43 L-methionine + CO2 + NH3
-
?
N-carbamoyl-DL-norleucine + H2O
-
Geobacillus stearothermophilus L-norleucine + CO2 + NH3
-
?
N-carbamoyl-DL-norvaline + H2O
-
Geobacillus stearothermophilus L-norvaline + CO2 + NH3
-
?
N-carbamoyl-DL-phenylglycine + H2O low activity Geobacillus stearothermophilus L-phenylglycine + CO2 + NH3
-
?
N-formyl-DL-2-aminobutyric acid + H2O
-
Geobacillus stearothermophilus L-2-aminobutyric acid + CO2
-
?
N-formyl-DL-ethionine + H2O
-
Geobacillus stearothermophilus L-ethionine + CO2
-
?
N-formyl-DL-homophenylalanine + H2O
-
Geobacillus stearothermophilus L-homophenylalanine + CO2
-
?
N-formyl-DL-methionine + H2O
-
Geobacillus stearothermophilus L-methionine + CO2
-
?
N-formyl-DL-methionine + H2O
-
Geobacillus stearothermophilus CECT43 L-methionine + CO2
-
?
N-formyl-DL-norleucine + H2O
-
Geobacillus stearothermophilus L-norleucine + CO2
-
?
N-formyl-DL-norvaline + H2O
-
Geobacillus stearothermophilus L-norvaline + CO2
-
?
N-formyl-DL-phenylglycine + H2O
-
Geobacillus stearothermophilus L-phenylglycine + CO2
-
?

Synonyms

Synonyms Comment Organism
BsLcar
-
Geobacillus stearothermophilus
L-N-carbamoylase
-
Geobacillus stearothermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45 70 biocatalyst system, optimum temperature ranges are 45-55°C for N-formyl-amino acids and 55-70°C for N-carbamoyl-derivatives Geobacillus stearothermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
pH 8.0, the bienzymatic system retains almost total operability after 24 h of incubation at 45°C, overview Geobacillus stearothermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
biocatalyst system for both N-formyl- and N-carbamoyl-amino acid substrates Geobacillus stearothermophilus

pH Stability

pH Stability pH Stability Maximum Comment Organism
8
-
45°C, the bienzymatic system retains almost total operability after 24 h of incubation at pH 8.0, overview Geobacillus stearothermophilus