Method for producing optically active N-protected-propargylglycine

A method for producing (S)-N-protected-propargylglycine of the following formula (2), which comprises asymmetrically hydrolyzing an N-protected-propargylglycine of the following formula (1): wherein R 1 represents an alkyl group having 1 to 4 carbon atoms, R 2 and R 3 each independently represent a hydrogen atom or a protective group of an amino group, R 2 and R 3 being not simultaneously a hydrogen atom, and a carbon atom appended with a mark * represents an asymmetric carbon atom, using an asymmetric hydrolysis enzyme derived from a microorganism selected from Thermomyces genus, Aspergillus genus, Rhizopus genus, Penicillium genus, Pseudomonas genus, Humicola genus, Burkholderia genus, Candida genus and Streptomyces genus, or a cultured substance of a microorganism having an ability of producing this enzyme or a treated substance thereof, or using an asymmetric hydrolysis enzyme which is ±-amylase derived from a microorganism selected from Bacillus subtilis and Bacillus licheniformis.