Lactobionic acid and its salts are substances that have several applications in the pharmaceutical area. These products were obtained by bioconversion through the enzymatic complex glucose-fructose-oxidoreductase (GFOR)/gluconolactonase (GL) present in calcium alginate immobilized cells of Zymomonas mobilis. In the reactions catalyzed by this enzyme system, the medium pH must be controlled at slightly acid values. For this purpose, NaOH, KOH, or Ca(OH)2 were used, and as a result, the respective salts were formed. The kinetic study on the formation of sodium, potassium and calcium lactobionates was followed by the steps of purification and characterization aiming the potential use of these compounds in the pharmaceutical area. In the assays for the bioproduction of sodium, potassium or calcium lactobionates, yields of 74, 77 and 84% were obtained, respectively. In the salts purification step, purity levels of approximately 95% were achieved. The structural identities of the lactobionate salts were determined by high resolution mass spectrometry, in addition to the 13C and 1H NMR analysis. The characterization demonstrates the selectivity of the enzymatic reaction of GFOR/GL of Z. mobilis, in the production of lactobionic acid and its salts.