Sugar content is one of the important parameters to determine the quality of banana. In this paper, a non-destruction measurement system for sugar content is introduced using hyperspectral camera system over the VIS/NIR (400-1,000 nm) spectral range. Hyperspectral image (HSI) calibration was performed to compute reflectance value of banana surface in full wavelength range while spectral and spatial analysis was conducted using a partial least squares regression (PLSR) to create a model that computing relationship between the HSI spectra and the sugar content. The ground truth value of sugar content was measured using digital refractometer on the extracted banana sample. The proposed system was evaluated using 90 Ambon bananas (Musa acuminata Colla) which consist of 30 raw, 30 mature and 30 overripe banana. The PLSR model provided the root mean square error of 0.79 % and the correlation coefficient R2 of 0.988 in the full wavelength band. Finally, the proposed non-destruction prediction system could be implemented as an instrument for sugar content measurement of banana fruit.