3-Methylthio-1-propanol (3-Met) is widely used as a flavoring substance and an essential aroma ingredient in many foods. Producing 3-Met by microbial transformation is green and eco-friendly. In the present study, one strain, YHM-G, which produced a high level of 3-Met, was isolated from the Baijiu-producing environment. Strain YHM-G was identified as Hyphopichia burtonii according to its morphological properties, physiological and biochemical characteristics, and ribosomal large subunit 26S rRNA gene D1/D2 domain sequence analysis. The optimal conditions for 3-Met production by YHM-G were obtained by single factor design, Plackett–Burman design, steepest ascent path design and response surface methodology as follows: 42.7 g/L glucose, pH 6, 0.9 g/L yeast extract, 6 g/L L-methionine (L-Met), culture temperature 28 °C, shaking speed 210 rpm, loading volume 50 mL/250 mL, inoculum size 0.5% (v/v), culturing period 48 h and 2.5 g/L Tween-80. Under these optimal conditions, the 3-Met production by strain YHM-G was 3.16 g/L, a value 88.1% higher than that before optimization. Strain YHM-G can also produce a variety of flavor compounds that are important for many foods. This strain thus has the potential to increase the abundance of 3-Met in some fermented foods and enhance their aroma profiles.