Gene cloning, purification, and characterization of α-methylserine aldolase from Bosea sp AJ110407 and its applicability for the enzymatic synthesis of α-methyl-L-serine and α-ethyl-L-serine

The gene encoding alpha-methylserine aldolase was isolated from Bosea sp. AJ110407. Sequence analysis revealed that the predicted amino acid sequence encoded by the 1320-bp open reading frame was 65.0% similar to the corresponding sequence of the enzyme isolated from Ralstonia sp. AJ110405. The gene was expressed in Escherichia coli, and the recombinant enzyme was purified. Gel filtration revealed the molecular mass of the purified enzyme to be approximately 78 kDa, suggesting that the enzyme is a homodimer. The enzyme exhibited a specific peak at 429 nm in the spectrum and contained I mol pyridoxal 5'-phosphate per mole of the subunit. The V-max value was 1.40 mu mol min(-1) mg(-1), and the K-m value was 1.5 mM for the reaction wherein formaldehyde was released from alpha-methyl-L-serine. This enzyme could also catalyze the reverse reaction, i.e., the synthesis of alpha-methyl-L-serine from L-alanine and formaldehyde. This activity was inhibited in the excess of formaldehyde; however, alpha-methyl-L-serine was efficiently produced from L-alanine in the presence of formaldehyde. This method was also applicable for producing alpha-ethyl-L-serine from L-2-aminobutyric acid. (C) 2008 Elsevier B.V. All rights reserved.