Regulation of process parameters is a cost-effective approach to control microorganism-derived dissolved organic nitrogen (mDON) formation in low-temperature biological wastewater conditions. However, the integrated influence of multiple parameters in this process is poorly defined. In this study, mathematical methodology was used to evaluate the combined effects of hydraulic retention time (HRT), solids retention time (SRT), and mixed liquor suspended solids (MLSS) on mDON formation at 8 °C. This study also systematically explored how multiple combinations of those three parameters affected mDON chemodiversity (fluorescent properties and molecular compositions), microbial compositions, and specific relationships between mDON molecules and microbial species in activated sludge systems. Results showed that combined effects significantly controlled the mDON formation at 8 °C (P < .05). The systematic analysis suggested that the multi-parameter effects modulated the distribution of different mDON compositions and shaped the microbial communities. Most bacterial phyla as the generalist and a few as the specialist were displayed in 2487 pairs of strong microbe-mDON connections (|r| ≥ 0.6, P < .05). Moreover, network analysis on microbe-mDON relationships identified the network centers as crucial media in terms of combined effects of process parameters on mDON formation. Our results provide comprehensive insight on the roles of multi-parameter covariation influences in regulating the high complexity of mDON traits and microbe-mDON linkages, thereby highlighting the necessity to focus on the combined effects of process parameters for effective and correct controlling strategies on mDON concentrations. Display omitted •Parameter covariations significantly control mDON formation at 8 °C (P < .05).•Multi-parameter combined effects include linear and interactive functions.•mDON molecular traits exhibit different responses to different parameter combinations.•Most phyla show high connectivity with mDON molecules as network centers.•Multi-parameter covariations enable various microbe-mDON relationships.