sulforaphane is a kind of isothiocyanate, which is obtained by hydrolysis of glucosinolate by the unique myrosinase in plants. It has been proved to prevent the occurrence of many chronic diseases, such as obesity, diabetes and cancer. The impact of SFN on obese human gut flora, however, has not been established. In this research, SFN was isolated from broccoli seeds and then refined to achieve 95% purity. Next, an investigation was conducted into the digestion and fermentation processes of SFN. The stability of the SFN in simulated saliva, gastric fluid, and intestinal juice provides evidence that it can reach the gut and be available for utilization by gut microflora. In vitro fermentation of SFN by gut microbes in obese patients results in alteration in constitution of microbiota and production of short chain fatty acids. As the result of SFN ingestion by human gut bacteria, the content of butyric and valeric acids increased 1.21- and 1.46-fold, respectively. In obese human guts, the relative abundances of the beneficial genera including Lactobacillus, Weissella, Leuconosto, Algiphilus and Faecalibacterium significantly increased, whilst the detrimental genera, such as Escherichia-Shigella, Klebsiella, Clostridium_sensu_stricto_1, Sutterella, Megamonas and Proteus drastically declined. Taken together, these findings demonstrate that SFN can be used as a nutraceutical ingredient for obese patients and for improving human health.