The invention provides a method for screening stereoselective alpha-hydroxy acid
dehydrogenase, comprising the following steps: (1) dissolving a sample to be detected containing alpha-hydroxy acid
dehydrogenase in a buffer with pH of 6-9, adding an alpha-hydroxy acid chiral
monomer as a substrate, and carrying out
conversion reaction in water-bath at 20-50 DEG C; and (2) after the
conversion reaction, adding the conversion solution in an FeCl3 developer solution to conduct color development reaction for 5-30 min, when the color development reaction finishes, judging the result according to the color appeared in the reaction solution. The method has no need to carry out
derivatization on the substrate which is intend to screen, can rapidly identify the
dehydrogenase activity and optical selectivity of the selected microbe by using simple colourimetry, has the advantages of simple screening flow,
fast speed, low request for devices, strong versatility and the like, and offers great conveniences to the obtainment of optically pure products by using racemic
mandelic acid and
related derivatives as substrates to carry out resolution through biological enzymatic method.