Biomass is a key factor in fermentation process, directly influencing the performance of the fermentation system as well as the quality and yield of the targeted product. Therefore, the on-line estimation of biomass is indispensable. The soft-sensor based on support vector machine (SVM) for an on-line biomass estimation was analyzed in detail, and the improved SVM called the weighted least squares support vector machine was presented to follow the dynamic feature of fermentation process. The model based on the modified SVM was developed and demonstrated using simulation experiments.