In this study, kaempferol and its rhamnosides (α-rhamnoisorobin, afzelin, kaempferitrin) were isolated electively by bioconversion technology from Hibiscus cannabinus L. leaves to evaluate the anti-wrinkles effects and anti-microbial effects. In order to evaluate anti-wrinkles activity, reduction of expression matrix metalloproteinase-1 (MMP-1) protein and proliferation/ pro-collagen production were investigated. Kaempferol and α-rhamnoisorobin showed inhibition activity of MMP-1 generated to compared to positive control. In HaCaT cell proliferation assay, kaempferol and α-rhamnoisorobin significantly promoted cell proliferation in a concentration-dependent manner. In addition, procollagen synthesis assays (by HDF-N cell) showd that TGF-β induced procollagen production and also, all four kinds of experimental significantly promoted procollagen synthesis in a concentration-dependent manner. Kaempferol and α-rhamnoisorobin exhibited strong antimicrobial activities on five of microbes, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger.