The invention discloses threonine aldolase, mutants, threonine aldolase and mutant coding genes, recombinant vectors constructed by coding genes, recombinant genetically engineered bacteria obtained by transformation of recombinant vectors, and threonine aldolase and the application of mutants in the preparation of 2- / 3- / 4-position substituted phenylserine derivatives. The present invention uses threonine aldolase and mutants as biocatalysts, 2- / 3- / 4-position substituted benzaldehyde as substrate, pyridoxal 5-phosphate as coenzyme, glycine / glycine ester as Auxiliary substrate, carry out enzyme-catalyzed reaction in appropriate conditions and media, separate and prepare a series of phenylserine derivatives with different substituents, the total yield of this method is in the range of 76-99%, the ee value of the product is greater than 99%, and the de value is greater than 99%. The range is 70‑99%.