Chiral recognition of l-amino acids is of significant importance due to the crucial role of l-amino acids in life sciences and pharmaceutics. In this work, a chiral sensor with capability of probing two chiral amino acids by an attractive single-template molecular imprinting strategy is introduced and used in the simultaneous chiral recognition of d/l-alanine (d/l-Ala) and d/l-tyrosine (d/l-Tyr). The assay relies on the hydrolysis of l-alanyl-l-tyrosine dipeptide doped in silica/polypyrrole (SiO2/PPy) under acidic conditions, resulting in l-Ala and l-Tyr coimprinted chiral sensor. This work opens up a new avenue for simultaneous chiral sensing of two or more chiral amino acids by incorporating only one template, circumventing the shortcomings encountered with multitemplate molecularly imprinted technology.