The invention relates to the technical field of preservative films, in particular to a novel biological preservative film and a preparation method thereof. The film is prepared from the following components in parts by weight: 10-90 parts of polylactic acid, 10-90 parts of biodegradable aliphatic-aromatic copolyester, 2-5 parts of a toughening agent, 1-3 parts of chitosan, 1-3 parts of an absorbent, 1-5 parts of antibacterial oil, 0.3-0.6 part of an antioxidant and 0.2-0.6 part of a lubricant. The biological preservative film disclosed by the invention can be applied to fresh keeping of various foods such as cooked foods, fruits and vegetables, meat and the like, has a strong bactericidal effect, can kill more than 650 bacteria within multiple minutes, has extremely strong antibacterial, antiseptic and fresh-keeping functions, also has a spontaneous modified atmosphere function, is convenient to use, does not need to use a fresh-keeping agent, and greatly prolongs the storage and fresh-keeping period of the food. Specifically, a common fresh-keeping film can only keep the food fresh for 2-3 days, but the preservative film can keep the food fresh for 15-30 days, which is 5-10 timesthat of the common fresh-keeping film.