Despite their common use as eating utensils in East Asia, chopsticks require complex fine motor-skills for adequate operation and are thus most frequently used with the dominant hand; however, the effect of training time on the proficiency of using chopsticks with the non-dominant hand, as well as the brain activity underlying changes in skill, remain unclear. This study characterised the effect of time spent training in chopstick operation with the non-dominant hand on chopstick-use proficiency and the related brain activity to obtain data that may help individuals who are obliged to change handedness due to neurological disease to learn to use their non-dominant hand in performing daily activities. Thirty-two healthy right-handed students were randomly allocated to training (n = 16) or control (n = 16) groups; the former received 6 weeks of training in chopstick use with their non-dominant (left) hand, and the latter received none. After training, significant improvements in the execution speed and smoothness of upper extremity joints were observed in the training group. Moreover, left dorsolateral prefrontal cortex activity significantly decreased, and bilateral premotor cortex activity significantly increased across training. These results indicated that 6 weeks of chopstick training with the non-dominant hand effectively improved chopstick operation.