This study was carried out to optimize dyeing conditions of unripe Citrus Unshiu extract on silk fabric and to evaluate antimicrobial activity of the dyed fabric for its potential use as a functional natural dye. Unripe fruits of Citrus Unshiu in Jeju Island, Korea, extracted in 80 % Ethanol solution to final solid dye powder were dyed on silk fabric under a variety of conditions such as dye bath concentrations, temperature, and dyeing duration together with mordanting. Dyeing fastness properties to washing, rubbing, perspiration, and light were tested and the antimicrobial activities of the dyed fabric against Staphylococcus aureus and Klebsiella pneumoniae were investigated quantitatively. As results, the fabric showed the maximum dye uptake (K/S) under the conditions of $80^{circ}C$ for temperature, 30 min for duration, and 600 % (o.w.f.) for dye concentration. Both pre- and post-mordanting seemed not to be effective on increasing K/S values of the dyed silk fabrics under the optimum dyeing conditions. Fastness ratings to washing, rubbing, and perspiration were all very good (4-5 grades) for both 300 and 600 % dyed silk fabrics. Excellent antimicrobial activities over 99 % reduction rate against two both bacteria were exhibited for all of dyed fabrics undergone more than 300 % of dye concentration. From these results, it was concluded that the dye concentration of 300 % of unripe Citrus Unshiu could be employed to produce antimicrobial silk fabric. Furthermore, to get more saturated shades on the fabric by the citrus, higher dye concentration such as 600 % was available as well.