In this study, we show the successful application of different model-based approaches for the maximizing of macrolactin D production by Paenibacillus polymyxa. After four initial cultivations, a family of nonlinear dynamic biological models was determined automatically and ranked by their respective Akaike Information Criterion (AIC). The best models were then used in a multi-model setup for robust product maximization. The experimental validation shows the highest product yield attained compared with the identification runs so far. In subsequent fermentations, the online measurements of CO2 concentration, base consumption, and near-infrared spectroscopy (NIR) were used for model improvement. After model extension using expert knowledge, a single superior model could be identified. Model-based state estimation with a sigma-point Kalman filter (SPKF) was based on online measurement data, and this improved model enabled nonlinear real-time product maximization. The optimization increased the macrolactin D production even further by 28% compared with the initial robust multi-model offline optimization.