Although the nutrient compositions of edible mushrooms are well-studied, the effect of combining different mushrooms on their anti-glycation and antioxidant activities remains unknown. This study therefore aimed to identify mushroom combinations that exhibit synergistic anti-glycation and antioxidant activities. Five edible mushroom species, namely Agaricus bisporus, Lentinula edodes, Pleurotus ostreatus, Pleurotus eryngii, and Flammulina velutipes, were evaluated both individually and in pairwise combinations. Their bioactive profile (phenolics, tannins, flavonoids, and polysaccharides), as well as antioxidant and anti-glycation activities were analyzed to determine the types of activity interaction: synergism, addition, or antagonism. A. bisporus (7.5 mg/mL) showed the highest reducing capacity and tannin content. L. edodes demonstrated the strongest radical scavenging potential, while F. velutipes displayed the highest anti-glycation activity and phenolic content. Despite its high polysaccharide level, P. eryngii showed low antioxidant activity. Pairwise combinations revealed synergistic anti-glycation and antioxidant effects at low sample concentrations, while antagonistic anti-glycation and antioxidant effects were observed at high sample concentrations. The mushrooms’ polyphenols, tannins, and flavonoids were positively correlated with their antioxidant activity (r = 0.325 to 0.825, p < 0.05). However, they showed an inverse relationship (r = –0.349 to –0.644, p < 0.05) with polysaccharides and anti-glycation activity. The principal component analysis revealed that the types of bioactive content and mushroom combinations contributed to respective 53 and 23% of total activity variances. The best-performing mushroom combinations with synergistic anti-glycation and antioxidant activities were the mixtures of 7.5 mg/mL A. bisporus + 15 mg/mL F. velutipes, 7.5 mg/mL L. edodes + 7.5 mg/mL F. velutipes, and 7.5 mg/mL L. edodes + 15 mg/mL F. velutipes.