Optimal culture conditions were characterized for production of crude biosurfactant of Bacillus subtilis JK-1. During incubation of B. subtilis JK-1, the bacterial growth pattern, changes of the surface tension at variable temperatures, pH and NaCl concentrations in bacterial culture medium were studied. The strain was able to grow and produce biosurfactant at $15-45^{circ}C$, in the pH range of 6-10, and at 0-10% (w/v) NaCl. In case, culture broth pH was gradually changed to neutral or weak alkaline. Optimal culture conditions for crude biosurfactant production were at $35^{circ}C$ and pH 7.0 after 48 h incubation and the surface tension of biosurfactant was 24.0 mN/m. Besides, as the concentration of NaCl was increased from 0 to 10% (w/v), the growth was decreased, pH of the culture broth was converted from weak alkaline to acidic, and the surface tension rised.