Abstract: Background and Objective: Glutamate is a non-essential amino acid and it improves the perception of the taste umami and serves as a building block of protein and physiological functions of the body. Increased use of glutamate in animal feed causes glutamate to rise globally. The aim of this study was to obtain the optimal conditions for glutamate production by Lactobacillus plantarum VM. Materials and Methods: Lactobacillus plantarum VM (L. plantarum VM) is a lactic acid bacteria originating from Minangkabau fermented foods and produces glutamate. The increased production of glutamate from Lactobacillus plantarum VM can be achieved by improving the nutrition and the growth environment of the bacteria. This study was designed in the form of a laboratory experiment protocol and was repeated 3 times. The variables measured in this study were the medium pH, temperature, incubation time, carbon source and nitrogen source. Results: The results of this study showed an optimum 5.5 pH (161.519 mg L–1), incubation time (36 h), temperature (36°C) (350.001 mg L–1), 11% glucose (566,535 mg L–1) and 0.5% peptone (680.525 mg L–1). Conclusion: Optimization of the initial pH of the media, incubation time, temperature, source C and source N can increase glutamate production.