Irritable bowel syndrome (IBS) is a multifactorial disorder of the gut-brain axis, characterized by visceral hypersensitivity (VH). Psychobiotics, through GABA synthesis, are good candidates to alleviate gastrointestinal discomfort. Here, we analyzed the GABA-producer Lactococcus lactis CNCM I-5388 as an active-enzyme postbiotic to relieve VH mediated by psychological stress. L. lactis CNCM I-5388 was inactivated by ethanol while maintaining its glutamate decarboxylase (GAD) activity. This EtOH-treated nonviable form was given daily orally for 1, 5, or 10 days to female Wistar rats in comparison with viable L. lactis CNCM I-5388 or vehicle. Visceral sensitivity was measured by electromyography before and after partial restraint stress (PRS). GABA was quantified in the stomach collected from rats and in the gastric compartment of TIM-1 human gut model in fed state. A daily treatment for 5 and 10 days by L. lactis CNCM I-5388 both in its viable and nonviable forms counteracted VH promoted by PRS. However, only viable L. lactis CNCM I-5388 tended to reduce VH after a single administration. After 5-day treatment, only under PRS conditions, the production of GABA within the stomach was enhanced in rats treated with viable or nonviable L. lactis CNCM I-5388. This increase was confirmed by using the TIM-1 human gut model. We found that a postbiotic with an active-GAD enzyme of L. lactis CNCM I-5388, similarly to its viable psychobiotic form, exerts anti-VH properties in an IBS-like rat model. These effects are associated with GABA production in the stomach where the low pH promotes GAD activity.