Bacteria exist as aggregates or in biofilms to help with adaptation and protection from environmental stresses. The hypothesis that is tested in this paper is that the relative presence of glucose in the media, at the beginning of the growth phase, influences the surface chemistry of the cell, which as a consequence reduces the tendency for the cells to interact and form aggregates. In this study, we used Escherichia coli (E. coli) MG1655 as a model organism and measured the change in the surface chemistry of cells harvested at different growth phases, which had been cultured in Luria-Bertani media with and without the addition of glucose, using potentiometric titration and infrared spectroscopy. Cells, cultivated with the additional supplement of glucose at the beginning of the growth phase, displayed a higher concentration of bacterial surface functional groups and a variation in outer membrane proteins. As a consequence, the tendency for cell-to-cell attachment was significantly reduced. Our findings therefore revealed that glucose limits aggregation in E. coli MG1655 by altering the concentration of functional groups from macromolecules present on the bacterial surface.