In this work, colorants extraction process from gromwell was studied for making powder form of colorants by solving the high viscosity problem of gromwell extracts. In order to do that, sugar extracted together with colorants must be pre-extracted. For sugar decomposition, gromwell roots were pretreated with various enzyme solutions. The total sugar content of pre-extract with enzyme solution was measured. Accordingly, the effects of enzyme type and pretreatment condition on sugar decomposition were investigated to find appropriate enzyme(amylase, hemicellulase, pectinase) and enzyme activity (100~1000unit), pre-extracted time(3~24hr). Color characteristics and dye uptake of dyed fabrics were evaluated. Gromwell colorants were assessed for their potential antimicrobial activities, which possibly expand their end use as functional pigments. The efficiency of removing sugar was increased in the order of hemicellulase, pectinase, amylase, H2O. Gromwell colorants powder yield was in the range of 4.4% to 9.8% depending on pretreatment enzyme. Gromwell colorants produced RP color on the silk and wool fabrics with good dye uptake. Antimicrobial activity of gromwell colorants will greatly increase its potentiality for applying as functional natural colorants in the future.