CreBC is a highly conserved two-component regulatory system (TCS) in several gram-negative bacteria, including Escherichia coli, Aeromonas spp., Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. CreD is a conserved gene that encodes a predicted inner-membrane protein and is located near the creBC loci. Activation of CreBC increases creD expression; therefore, creD expression is generally used as a measure of CreBC activation in E. coli, Aeromonas spp., and P. aeruginosa systems. In this article, we aim to elucidate the expression of creD and further to investigate its functions in S. maltophilia. In spite of a short intergenic region of 81 bp between creBC and creD, creD is expressed separately from the adjacent creBC operon and from a promoter immediately upstream of creD (PcreD) in S. maltophilia. We found that the promoter activity of PcreD is negatively regulated by the creBC TCS, positively regulated by the bacterial culture density, and not affected by β-lactams. Furthermore, creD expression is not significantly altered in the presence of the phosphor-mimic variant of CreB, CreB(D55E), which mimics activated CreB. The functions of CreD of S. maltophilia were assessed by comparison among the following: wild-type KJ; the creD isogenic mutant, KJΔCreD; and the complementary strain, KJΔCreD(pCreD). The mutant lacking creD had cell division defects and aberrations in cell envelope integrity, which then triggered the σE-mediated envelope stress response. Thus, the results indicated that CreD plays a critical role in the maintenance of envelope integrity.