The estimation of source information from limited measurements of a sensor network is a challenging inverse problem, which can be viewed as an assimilation process of the observed concentration data and the predicted concentration data. When dealing with releases in built-up areas, the predicted data are generally obtained by the Reynolds-averaged Navier-Stokes (BANS) equations, which yields building-resolving results; however, RANS-based models are outperformed by large-eddy simulation (LES) in the predictions of both airflow and dispersion. Therefore, it is important to explore the possibility of improving the estimation of the source parameters by using the LES approach. In this paper, a novel source term estimation method is proposed based on LES approach using Bayesian inference. The source-receptor relationship is obtained by solving the adjoint equations constructed using the time-averaged flow field simulated by the LES approach based on the gradient diffusion hypothesis. A wind tunnel experiment with a constant point source downwind of a single building model is used to evaluate the performance of the proposed method, which is compared with that of the existing method using a BANS model. The results show that the proposed method reduces the errors of source location and releasing strength by 77% and 28%, respectively.