Listeria monocytogenes, a food-borne bacterial pathogen causing significant human mortality, propagates by expressing genes in response to environmental signals, such as temperature and pH. Listeria gene (lmo0799) encodes a protein homologous to the Bacillus subtilis YtvA, which has a flavin-light, oxygen or voltage (LOV) domain and a Sulfate Transporters Anti-Sigma factor antagonist (STAS) output domain that regulates transcription-initiation factor Sigma B in the bacterial stress response upon exposure to light. This could be significant for the pathogenesis of listeriosis because Sigma B has been linked to virulence of Listeria, and the Listeria Lmo0799 protein has recently been identified as a virulence factor activated by blue light. We have cloned, expressed heterologously in Escherichia coli and purified the full-length LM-LOV-STAS protein. Although it exhibits photochemical activity similar to that of YtvA, LM-LOV-STAS lacks an almost universally conserved arginine in the flavin-binding site, as well as another positively charged residue, a lysine in YtvA. The absence of these positive charges was found to destabilize retention of the flavin mononucleotide (FMN) chromophore in the LM-LOV-STAS protein, particularly at higher temperatures. The unusual sequence of the LM-LOV-STAS protein alters both spectral features and activation/deactivation kinetics, potentially expanding the sensory capacity of this LOV domain, e.g. to detect light plus cold.