The marine bacterium Vibrio parahaemolyticus is responsible for the seafood-borne gastroenteritis in humans. Two-component system (TCS) TtrRS plays important roles in host colonization and works as an important regulator in V. parahaemolyticus. However, the mechanisms by which TtrRS regulates the downstream genes, as well as the function of their target genes, remain largely unknown. In this study, we performed a Find Individual Motif Occurrences (FIMO) analysis using the ttrR box to scan the V. parahaemolyticus genome and identified 68 potential binding sites harboring the ttrR box. Thereinto, a ttrR box exists in the promoter region of the 05890-05900 operon. We further demonstrated that TtrR directly regulates the expression of the 05890-05900 operon by binding to the ttrR box. In addition, another TCS, 05905/05910, could assist in the regulation of TtrR to the 05890-05900 operon. Subsequent study showed that the 05890-05900 operon promotes the sulfur metabolism of V. parahaemolyticus at the early growth stage. Our results suggested that TtrRS and their target genes generate a complex regulatory pathway to better control sulfur metabolism in V. parahaemolyticus.Please check and confirm the edit made in the article title and amend if necessary.It's OK.