The invention discloses a method for preparing (S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid through multienzyme coupling. The method comprises the following steps: by taking raceme of 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid or raceme of 1,2,3,4-tetrahydroisoquinoline-3-formate as a substrate, enabling (R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid in the substrate to react to generate imidic acid shown in a formula (II) under the catalysis action of oxidative dehydrogenase; and converting the imidic acid shown in the formula (II) into the (S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid in the presence of piperidine acid reductase and a coenzyme capable of providing hydride ions. The method is gentle in reaction condition, good in stereoselectivity, high in reaction efficiency, high in conversion rate, and the like.