Production of 2-phenylethanol by Pichia fermentans L-5 as affected by cultivation temperature (20–40°C), medium pH (4.0–11.0) and shaking speed (0–250 rpm) as well as the kinetics of 2-phenylethanol formation during fermentation were investigated in the present study. Results showed the optimum culture temperature, initial medium pH and shaking speed for the 2-phenylethanol production by test organism were 30–35°C, 8.5 and 250 rpm, respectively. Kinetics study revealed that production of 2-phenylethanol during P. fermentans L-5 fermentation increased almost in parallel with the increase in biomass or viable population during the initial stage of fermentation. During a 72-h fermentation period, level of phenylalanine in medium dropped coincidentally with increasing biomass and 2-phenylethyl derivatives, from the start of fermentation. A maximum 2-phenylethanol production of ca 505.5 mg/l with a molar yield of ca 0.74 was obtained after 16 h of fermentation.