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Before the absorption in the intestine, glucose encounters gut bacteria, which may serve as a barrier against hyperglycemia by metabolizing glucose. In the present study, we compared the capacity of enterobacterial strains to lower glucose levels in an in vitro model of nutrient-induced bacterial growth. Two probiotic strains, <i>Hafnia alvei HA4597</i> (<i>H. alvei</i>) and <i>Escherichia coli</i> (<i>E. coli</i>) <i>Nissle 1917</i>, as well as <i>E. coli K12</i>, were studied. To mimic bacterial growth in the gut, a planktonic culture was supplemented twice daily by the Luria Bertani milieu with or without 0.5% glucose. Repeated nutrient provision resulted in the incremental growth of bacteria. However, in the presence of glucose, the maximal growth of both strains of <i>E. coli</i> but not of <i>H. alvei</i> was inhibited. When glucose was added to the culture medium, a continuous decrease in its concentration was observed during each feeding phase. At its highest density, <i>H. alvei</i> displayed more efficient glucose consumption accompanied by a more pronounced downregulation of glucose transporters’ expression than <i>E. coli K12</i>. Thus, the study reveals that the probiotic strain <i>H. alvei HA4597</i> is more resilient to maintain its growth than <i>E. coli</i> in the presence of 0.5% glucose accompanied by more efficient glucose consumption. This experimental approach offers a new strategy for the identification of probiotics with increased glucose metabolizing capacities potentially useful for the prevention and co-treatment of type 2 diabetes.