The invention provides pine pollen enzyme, a preparation method thereof, as well as enzyme jelly prepared from the pine pollen enzyme and a preparation method of the enzyme jelly. The pine pollen enzyme is mainly prepared from the following raw materials: pine pollen, pine nuts, edible flowers, edible fruits and vegetables, brown sugar, rock candy, an activated yeast liquid, a fermenting strain and water. The preparation method of the pine pollen enzyme comprises the following steps: (1) washing the pine pollen, pine nuts, edible flowers and edible fruits and vegetables, performing blanching,cooling and draining, and performing grinding with water, pulping and coarse filtering; (2) adding the brown sugar and rock candy into the mixture, and performing dissolving; (3) adding the activatedyeast liquid into the mixture, and performing fermentation; and (4) inoculating the mixture with the fermenting strain, and performing fermentation and post-fermentation. The enzyme jelly is mainly prepared from the following raw materials: the pine pollen enzyme, white granulated sugar, gelatin, agar, citric acid and water. The invention further discloses a method for preparing the enzyme jelly.The pine pollen enzyme has golden bright color, properly sour and sweet taste and excellent flavor; the enzyme jelly has a sour-sweet delicious taste and the enzyme flavor; and the methods are simple,have low cost, and are suitable for industrial production.