Effects of Sulfide Flavors on AHL‐Mediated Quorum Sensing and Biofilm Formation of Hafnia alvei

Abstract In this study, 10 different sulfide flavor compounds commonly used as food additives were screened for antiquorum‐sensing activity. Among these, diallyl disulfide (DADS) and methyl 2‐methyl‐3‐furyl disulfide (MMFDS) were found to exert the strongest inhibition against violacein production in Chromobacterium violaceum 026, the tested biosensor strain. DADS and MMFDS also inhibited the growth of Hafnia alvei H4, yielding MIC values of 48 and 41.6 mM, respectively. In addition, DADS and MMFDS also inhibited the ability of H. alvei H4 to produce acyl‐homoserine lactone as demonstrated by the reduced level of C6‐HSL in the supernatant of DADS‐treated culture. At concentrations corresponding to 1/4 MIC, DADS, and MMFDS inhibited the swarming ability of H. alvei H4 by 73.50% and 76.43%, respectively, while having virtually no effect on cell growth. The same concentrations of DADS and MMFDS also completely inhibited the formation of biofilm. These antiquorum sensing effects of DADS and MMFDS involved changes in the expression of the quorum‐sensing genes luxI and luxR . Quantitative RT‐PCR analysis showed that the mRNA levels of both genes were significantly reduced by DADS and MMDFS at concentrations below their MICs. However, further test using a mutant strain of H . alvei lacking luxR ( ΔluxR ) revealed significant reduction in luxI mRNA level upon treatment of the strain with DADS or MMDFS, but no change in luxR mRNA level occurred when a luxI ‐lacking mutant ( ΔluxI ) was treated with these compounds. The result therefore suggested that the antiquorum‐sensing effect of DADS and MMFDS against H. alvei H4 might operate mainly through the inhibition of luxI expression in the cells. Practical Application The sulfide flavors compounds used in this paper are commonly used in food processing in China and are listed in the national standard of Chinese food additives GB2760‐2014. The application of sulfide flavors in food processing can enhance aroma and prevent food spoilage.