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ChemicalBook CAS DataBase List DL-3-Aminoisobutyric acid

DL-3-Aminoisobutyric acid synthesis

10synthesis methods
-

Yield:-

Reaction Conditions:

with phosphoric acid;dihydropyrimidinase;N-carbamoyl- -alanine amidohydrolase;nickel dichloride in aq. phosphate buffer at 30; pH=8;Enzymatic reaction;

Steps:

2.6. Bienzymatic system characterization

To analyze the effect of Ni2+ on enzymatic activity, samples of purified At car and SmelDhp enzymes were incubated together in the presence of different concentrations of NiCl2 at 4 ?C for several days. Standard enzymatic reaction was carried out with (R,S)-5-METDHU as substrate (10 mM) in 100 mM sodium phosphate buffer (pH 8.0), at 30 ?C in 1 ml reaction volume. Aliquots of 100 l were taken and stopped by addition of 900 l of 1% H3PO4. After centrifuging, the resulting supernatants were analyzed by high-performance liquid chromatography (HPLC). The HPLC system (LC2000Plus HPLC System, Jasco, Madrid, Spain) equipped with a Luna C18 column (4.6×250 mm; Phenomenex, Madrid, Spain) was used to determine the concentrations of 5-METDHU, N-carbamoyl-3-AiBA, and 3-AiBA. The mobile phase used in the analysis was 95% phosphoric acid (20 mM, pH 3.2) and 5% methanol, pumped at a flow rate of 1 ml min-1. The UV detector was fixed at 200 nm. All reactions conducted for the bienzymatic system characterization were carried out in triplicate. Optimal temperature of the bienzymatic system was evaluated from 25 to 60 ?C. Thermal stability was measured after 24 h of preincubation at temperatures from 4 to 50 ?C. A pH range of 6.0-9.0 was assayed (sodium phosphate and Tris/HCl buffers), at a concentration of 100 mM. Standard enzymatic reaction was then carried out with the Ni-amended At car (1 M) and SmelDhp (1 M) enzymes. The reaction progress was monitored by HPLC analysis as described above.

References:

Martínez-Gómez, Ana Isabel;Clemente-Jiménez, Josefa María;Rodríguez-Vico, Felipe;Kanerva, Liisa T.;Li, Xiang-Guo;Heras-Vázquez, Francisco Javier Las;Martínez-Rodríguez, Sergio [Process Biochemistry,2012,vol. 47,# 12,p. 2090 - 2096]

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