The tyrosinase inhibitor producing strain, Burkholderia cepacia TKU025 was isolated from the soil in the northern Taiwan by using squid pen powder (SPP) as the sole carbon/nitrogen source and identified as the Burkholderia cepacia. The strain was a non-lactose-fermenting and Gram-negative bacteria. The optimal conditions for tyrosinase inhibitor production were cells used to inoculate in 25 mL of medium containing 1.0% SPP, 0.1% K2HPO4 and 0.05% MgSO4．7H2O（pH7）and shaken at 30 °C for 4 days. The tyrosinase inhibitor was purified by ethyl acetate, chromatography of Sepharose S-100 and thin-layer chromatography (tlc). The tyrosinase inhibitor had broad thermal stability and pH stability. It maintained it’s initial activity at 100 °C and pH 2-12, respectively. Besides, the purified inhibitor by Sepharose S-100 chromatography had antimicrobial activity against E. coli and B. cereusTKU006. Both the best inhibition were more than 95 %.