Introducing functional metal nanoparticles (NPs) into flexible substrate is being increasingly attempted to expand their application. Here, we extend the synthesis of cellulose to its unmodified dope achieving freestanding nanocomposite decorated with bimetallic Ag-Au NPs through the one pot reaction. In the procedure, cellulose chain not only acts as a reducing agent but also a biocompatible support for NPs with a mean size of 7.9-9.7 nm. Meanwhile, changing the addition order of Ag and AuCl generated different atom arrangement in the bimetallic NPs. Moreover, the correlation of bioactivity to NP atom arrangement was studied. The result revealed that the nanocomposite containing NPs with an ultrathin Ag-rich outermost shell around an Au-rich core showed better bactericidal ability while lower cytotoxicity. In addition, the nanocomposite exhibited a sensitive SERS property for determination of R6G with a high enhancement factor of 10.