In this work, electrochemical methods were used to prepare surface-enhanced Raman scattering (SERS)-active gold substrates to investigate the effect of substrate temperatures on improved SERS performances. The results indicate that the SERS enhancement capabilities are gradually raised from 25°C to a maximum at 40°C and monotonically decreased from 40 to 100°C. The SERS of rhodamine 6G (R6G) adsorbed on the SERS-active substrate at 40°C exhibits a higher intensity by 4-fold of magnitude, as compared with that of R6G adsorbed on the SERS-active substrate at 25°C. Also, SERS of polypyrrole (PPy) deposited on the roughened Au substrate treated at 35°C exhibits the highest intensity by 3-fold of magnitude, as compared with that of PPy deposited on the same roughened Au substrate at room temperature.