Propolis is a promising natural product that has been extensively researched and studied for its potential health and medical benefits. The lack of requisite high oil-containing propolis and existing variation in the quality and quantity of essential oil within agro-climatic regions pose a problem in the commercialization of essential oil. As a result, the current study was carried out to optimize and estimate the essential oil yield of propolis. The essential oil data of 62 propolis samples from ten agro-climatic areas of Odisha, as well as an investigation of their soil and environmental parameters, were used to construct an artificial neural network (ANN) based prediction model. The influential predictors were determined using Garson’s algorithm. To understand how the variables interact and to determine the optimum value of each variable for the greatest response, the response surface curves were plotted. The results revealed that the most suited model was multilayer-feed-forward neural networks with an R2value of 0.93. According to the model, altitude was found to have a very strong influence on response, followed by phosphorous & maximum average temperature. This research shows that using an ANN-based prediction model with a response surface methodology technique to estimate oil yield at a new site and maximize propolis oil yield at a specific site by adjusting variable parameters is a viable commercial option. To our knowledge, this is the first report on the development of a model to optimize and estimate the essential oil yield of propolis.