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Aims: Development of cost‐effective production medium by applying statistical designs for single‐step fermentation of starch (corn flour – CF) to l‐(+) lactic acid, using inexpensive nitrogen sources as substitutes for peptone and yeast extract in MRS medium by amylolytic Lactobacillus amylophilus GV6.

Methods and Results: A two‐level Plackett–Burman design was employed for screening various available crude starches (flours) for l‐(+) lactic acid production by Lact. amylophilus GV6 using red lentil flour (RL) and bakers yeast cells (YC) as substitutes for commercial peptone and yeast extract in MRS medium in anaerobic submerged fermentation. Of all the tested flours, CF was found to be the most significant. Central composite rotatable design was employed to determine maximum production of l‐(+) lactic acid at optimum values of process variables, CF, RL, YC, CaCO₃ and incubation …