Various disorders closely linked to gut dysbiosis have been associated with poor dietary patterns. Dietary prebiotic fibers play an essential role in modulating the gut microbiome by enhancing the abundance of beneficial microorganisms and improving the production of short-chain fatty acids. Arabinoxylan (AX) is a major component of most dietary fibers and has been shown to exhibit potential prebiotic properties and modulate gut microbiome composition. This study aimed to investigate the in vitro impact of bioengineered wheat arabinoxylan on depression-inversely linked gut microbes and human gut microbiome diversity and metabolism. This study demonstrates the ability of bioengineered AX to stimulate the growth of depression-inversely linked gut bacterial species (Faecalibacterium prausnitzii and Lacticaseibacillus rhamnosus LGG). On the microbiome composition, the bioengineered AX induced an increased abundance of beneficial bacterial taxa (Bacteroides, Bifidobacterium, Anaerofustis, and Eubacterium) compared to the control and native AX. These effects on microbes translated into significant metabolic activity and produced primary SCFAs (acetate, butyrate, and propionate). The findings from this study suggest that bioengineered wheat arabinoxylan could be considered a promising strategy for fostering next-generation prebiotics targeting depression-inversely linked gut microbes and also supports the structure-function relationship between AX and the human gut microbiome.