Brain bioenergetics could be affected by many intrinsic and extrinsic factors, yet no link has been established between markers of bioenergetics in specific brain regions and educational level. In this cross-sectional study, we evaluated N-acetylaspartate-to-creatine ratio (NAA/Cr), a biomarker of brain functionality, using multivoxel proton magnetic resonance spectroscopy in twelve brain locations among healthy volunteers (n = 51; 30 females; age 47.8 ± 16.3 years) who earned either a high school diploma (36 out of 51) or a college degree (15 out of 51). Hierarchical multiple regression analysis revealed that our model as a whole (including level of education as a predictor variable, and age and gender as control variables) explained significant percentage of the variance in the NAA/Cr levels at 8 out of 12 specific brain locations (P < 0.05). This was highlighted in left anterior mesial cortex where the model explained 63.1% of the variance in brain NAA/Cr, with education and age make significant contributions (72.6% and 48.5%, respectively) to our model (P < 0.05). Having superior brain N-acetylaspartate-to-creatine ratio appears to be related with higher education in healthy men and women.