Sugarcane juice is a highly perishable beverage, and its stabilization may be achieved by applying the proper heat treatment. This study aimed at optimizing the binomial holding time x temperature for whole cane juice pasteurization. Eleven runs were carried out according to a Central Composite Rotatable Design (CCRD). The pasteurized juice was aseptically filled into polyethylene terephthalate (PET) bottles and stored at 0 °C in the dark. The temperature (ranging from 78 to 92 ºC) and holding time (from 16 to 44 s) were tested as independent variables. The performances of the binomials were evaluated by physicochemical, microbiological, enzymatic and sensory tests, and instrumental measurement of color. The data were processed through Analysis of Variance (ANOVA), Tukey test, Principal Component Analysis (PCA), and Response Surface. The values obtained for pH, soluble solids content (SS) and titratable acidity (TA) of freshly extracted juice ranged from 5.15 to 5.37; 17.8 at 24.5 °Brix; and 0.052 to 0.125% (citric acid), respectively. The pasteurized juice showed values of 5.18 to 5.37 for pH, 17.4 to 24.1 for TSS and 0.062 to 0.123% for TA. The eleven samples, produced in different runs, showed significant differences in terms of physicochemical parameters. The comparison between the samples of fresh and pasteurized juice indicated that the heat treatment had little or no influence on those parameters. The lightness variations (ΔL *), red / green chroma (Δa *) and yellow / blue chroma (Δb *) ranged from-2.3 to +2.4;-3.2 to-0.9; and-5.5 to +0.2, respectively. The total color difference (ΔE *) between fresh and pasteurized juice ranged from 1.95 to 5.91. Decimal reductions of microorganisms ranged from 2.9 to > 4.7 for mesophiles, > 2.9 to > 3.8 for molds and yeasts, and > 3.0 to > 4.7 for psychrotrophics. Reductions in enzymatic activity ranged from 24.6 to 89.5% for polyphenoloxidase (PPO) and from 47.8 to 93.7% for peroxidase (POD). The average scores obtained for the pasteurized juice ranged from 5.6 to 7.3 for appearance, from 7.2 to 7.8 for flavor and from 6.9 to 7.7 for overall impression. The impact of the binomials applied to the pasteurization of the sugarcane juice widely varied. With respect to enzymatic inactivation and microorganism's reduction, the treatment at 90 ºC/40 s was the most efficient. The treatment at 85 ºC/44 s was the most effective in maintaining the original color of juice. In terms of sensory acceptability, the treatment at 85 ºC/16 s had the best performance. The binomial 90 ºC / 40 s was considered the most suitable for sugarcane juice pasteurization.