4 types of lactic acid bacteria (Lactobacillus bulgaricus, L. acidophilus, L. casei, and L. casei) were selected in this study. First, the zinc-rich culture process of lactic acid bacteria was optimized, and then the antioxidant activity of the fermentation supernatant of them was measured, and the digestion stability and bioavailability were further analyzed by in vitro simulated digestion. The results showed that the optimal of zinc addition time for zinc-rich lactic acid bacteria was 10 hours after inoculation, and the zinc ion concentration was 50 μg/mL. The antioxidant activity of lactic acid bacteria supernatant could be improved by zinc-rich culture. Four different types of lactic acid bacteria cultured with oyster polypeptide zinc had higher supernatant free radical scavenging activity than that from other zinc sources. The gastrointestinal release rate of L. bulgaricus cultured with oyster peptide-zinc chelates was (3.46±0.17)%, and the bioavailability was the highest (29.78±0.42) %. In summary, L. bulgaricus was a potential dominant zinc-rich strain. L. bulgaricus cultured with the zinc-rich zinc oyster peptide had good in vitro antioxidant activity, gastrointestinal digestive stability, and bioavailability.