The invention discloses a biological brewing method for high-quality mature honey. The method comprises the following steps: firstly a honey-brewing room is constructed, which is composed of at leastone closed room with a staff entrance door, and beehives are placed along the wall in the honey-brewing room, the wall of the honey-brewing room corresponding to the position of a beehive door is provided with a bee access hole communicating with the external environment; a filling barrel is placed outside the honey-brewing room or inside the honey-brewing room, and a conveying pipe hermetically connected sealed with the filling barrel extends to inside of the beehive; the immature honey is poured into the barrel, the immature honey is conveyed into the beehive through the conveying pipe to feed the bee until the honeycomb billet is completely sealed with the beeswax cover. The method is simple and easy to grasp, and the use of biological brewing greatly improves the quality and taste of the honey, because of the indoor environment, the floral fragrance of honey will not be emitted, the phenomenon of 'stealing bees' is eliminated, at the same time, the problem that bees only eat but donot work during the short period of the honey source is also solved which ensures the continuous production of honey and reduces the production costs of beekeepers.