Mushrooms have limited shelf-life and it can be prolonged if suitable conditions and treatments are effectively applied. In this study, nanocomposite material and antimicrobial agents with a combination of chitosan were used as novel packaging material for mushroom preservation. The microbiological analysis, physicochemical properties, headspace gas analysis, and polyphenol oxidase activity (PPO) during cold storage were investigated. As compared with control, coated mushrooms with chitosan (CHS), and nano-titanium dioxide CHSTiO2 thymol + tween-80 CHSTiO2/TT80 coating treatment showed significantly (p ≤ 0.05) lower respiration rate, microbial contaminations (4.27 log CFU/g), and (5.93 log CFU/g) for total yeast/mold and aerobic plate counts, respectively. The weight loss ratio was the lowest for CHSTiO2/TT80 (10.88% loss) followed by CHSTiO2 (11.76% loss). CHSTiO2/TT80 recorded a higher electrolyte leakage rate (25.84%) and acidity. While the lowest PPO activity was established for CHSTiO2 (17.09 U mg−1 Protein), while the lowest values for total soluble solid concentrations were reported for CHSTiO2/TT80 mushrooms (4.91%). These results indicated that CHSTiO2/TT80 coating treatment might delay the aging degree of white button mushrooms and be investigated as a novel packaging material for other food products in the future.