l-DOPA (3,4-dihydroxyphenyl-l-alanine) is an extensively used drug for the treatment of Parkinson's disease. In the present study, optimization of nutritional parameters influencing l-DOPA production was attempted using the response surface methodology (RSM) from Brevundimonas sp. SGJ. A Plackett–Burman design was used for screening of critical components, while further optimization was carried out using the Box–Behnken design. The optimized levels of factors predicted by the model were pH 5.02, 1.549 g l−1 tryptone, 4.207 g l−1 l-tyrosine and 0.0369 g l−1 CuSO4, which resulted in highest l-DOPA yield of 3.359 g l−1. The optimization of medium using RSM resulted in a 8.355-fold increase in the yield of l-DOPA. The anova showed a significant R2 value (0.9667), model F-value (29.068) and probability (0.001), with insignificant lack of fit. The highest tyrosinase activity observed was 2471 U mg−1 at the 18th hour of the incubation period with dry cell weight of 0.711 g l−1. l-DOPA production was confirmed by HPTLC, HPLC and GC-MS analysis. Thus, Brevundimonas sp. SGJ has the potential to be a new source for the production of l-DOPA.