Effects of D-sorbitol concentration on the kinetics of D-sorbitol microbial oxidation were studied. The culture of Gluconobacter suboxydans S–22 was applied as a biocatalyst. The study was focused on the determination of oxygen uptake rates and the evaluation of factors affecting them. Special emphasis was given to estimations of dissolved oxygen saturation (Monod) constant (KsDO) and critical dissolved oxygen concentration. The estimated values of Monod constant (KsDO) were correlated with different process kinetics parameters and a series of acceptable relationships was established. Excellent correlation coefficients were found for KsDO as a linear function of (i) reciprocal viscosity (R2 = 0.9848), (ii) relative diffusion coefficient (R2 = 0.9983), (iii) cubic root of dissolved D-sorbitol mass concentration (R2 = 0.9915) and (iv) oxygen solubility in D-sorbitol solutions of different D-sorbitol concentrations (R2 = 0.9967). The estimated KsDO values were lower than their corresponding critical dissolved oxygen concentrations.