Efficient Preparation of Optically Active p-Trimethylsilylphenylalanine by Using Cell-Free Extract of Blastobacter sp. A17p-4

概要

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Optically active p-trimethylsilylphenylalanine (TMS-Phe) was prepared by enantioselective hydrolysis of N-carbbamoyl-DL-p-trimethylsilylphenylalanine (C-DL-TMS-Phe) with the cell-free extract of Blastobacter sp. A17p-4,which is known to produce N-carbamoyl-D-amino acid amidohydrolase (DCase). Although this bacterium also produced N-carbamoyl-L-amino acid aminohydrolase (LCase), heat treatment of the cell-free extract for 40 min at 50℃ and pH 7.0 was found to be effective in compoletely inactivating the LCase without loss of DCase activity, which provided a far simpler and more convenient method of preparing optically pure D-TMS Phe (99% enantiomeric excess, ee). Furthermore, optically pure L-TMS-Phe (99% ee) could be obtained by LCase-catalyzed hydrolysis of the residual substrate with the non-treated cell-free extract as the enzyme source. The optimum pH for the hydrolysis of C-L-TMS-Phe was 7.0,and addition of 2 mM Mn^<2+> and 5% N, N-dimethylformamide were effective in accelerating the activity of LCase and raising the substrate concentration, respectively.
公益社団法人日本生物工学会の論文
1997-09-25